Uterus Of Dairy Cows Research Articles

Pangenomic and biochemical analyses of Helcococcus ovis reveal widespread tetracycline resistance and a novel bacterial species, Helcococcus bovis.

Helcococcus ovis (H. ovis) is an opportunistic bacterial pathogen of a wide range of animal hosts including domestic ruminants, swine, avians, and humans. In this study, we sequenced the genomes of 35 Helcococcus sp. clinical isolates from the uterus of dairy cows and explored their antimicrobial resistance and biochemical phenotypes in vitro. Phylogenetic and average nucleotide identity analyses classified four Helcococcus isolates within a cryptic clade representing an undescribed species, for which we propose the name Helcococcus bovis sp. nov. By establishing this new species clade, we also resolve the longstanding question of the classification of the Tongji strain responsible for a confirmed human conjunctival infection. This strain did not neatly fit into H. ovis and is instead a member of H. bovis. We applied whole genome comparative analyses to explore the pangenome, resistome, virulome, and taxonomic diversity of the remaining 31 H. ovis isolates. An overwhelming 97% of H. ovis strains (30 out of 31) harbor mobile tetracycline resistance genes and displayed significantly increased minimum inhibitory concentrations of tetracyclines in vitro. The high prevalence of mobile tetracycline resistance genes makes H. ovis a significant antimicrobial resistance gene reservoir in our food chain. Finally, the phylogenetic distribution of co-occurring high-virulence determinant genes of H. ovis across unlinked and distant loci highlights an instance of convergent gene loss in the species. In summary, this study showed that mobile genetic element-mediated tetracycline resistance is widespread in H. ovis, and that there is evidence of co-occurring virulence factors across clades suggesting convergent gene loss in the species. Finally, we introduced a novel Helcococcus species closely related to H. ovis, called H. bovis sp. nov., which has been reported to cause infection in humans.

Dynamics and Diversity of Intrauterine Anaerobic Microbiota in Dairy Cows with Clinical and Subclinical Endometritis

The aim of the study was to characterize the dynamics of anaerobic cultivable postpartum microbiota in the uterus of dairy cows. In total, 122 dairy cows were enrolled and sampled on day 0 (day of calving) and on days 3, 9, 15, 21, and 28 postpartum (pp). Samples were cultivated anaerobically and analyzed by MALDI-TOF MS. In total, 1858 isolates were recovered. The most prevalent facultative anaerobic genera were Trueperella (27.8%), Streptococcus (25.4%), and Escherichia (13.1%). The most prevalent obligate anaerobes were Peptoniphilus (9.3%), Bacteroides (3.3%), and Clostridium (2.4%). The microbial communities were highly dynamic and diverse. On the animal level, Trueperella pyogenes on day 21 and 28 pp was associated with clinical endometritis, and E. coli on day 21 pp was associated with subclinical endometritis. The occurrence of Streptococcus pluranimalium on day 28 was related to uterine health. The presence of T. pyogenes, Streptococcus, and Peptoniphilus was significantly associated with an increased risk for purulent vaginal discharge. Primiparous cows showed a higher prevalence of T. pyogenes, Fusobacterium necrophorum, Porphyromonas levii, and Peptoniphilus spp. than multiparous cows but were not more susceptible to uterine diseases. This study might provide a suitable basis for future co-cultivation studies to elucidate potential synergistic interactions between microbiota.

The effects of endometrial epithelium-derived exosomes on the recruitment and activation of T Lymphocytes in the uteri of dairy cows

The effects of endometrial epithelium-derived exosomes on the recruitment and activation of T Lymphocytes in the uteri of dairy cows

Draft Genome Sequences of Helcococcus ovis Strains Isolated at Time of Metritis Diagnosis from the Uterus of Holstein Dairy Cows.

Helcococcus ovis is an emerging pathogen implicated in the pathogenesis of metritis in dairy cows. Herein, we report the first draft genome sequences of four Helcococcus ovis isolates from the uterus of dairy cows with metritis. This information will enable a better understanding of the bacterium's pathogenicity and antimicrobial resistance.

Draft Genome Sequences of Two Fusobacterium necrophorum Strains Isolated from the Uterus of Dairy Cows with Metritis.

A commensal in the gastrointestinal tract, Fusobacterium necrophorum is involved in the pathogenicity of abscesses, foot rot, and metritis in cattle. Here, we present the draft genomes of two Fusobacterium necrophorum isolates from the uterus of dairy cows with metritis to allow for future comparative genome studies.

Randomized clinical trial of ceftiofur hydrochloride treatment to prevent acute puerperal metritis in dairy cows having Escherichia coli in their uterus at day 1 postpartum

Ceftiofur is a class 1 antibiotic (World Health Organization classification) and there is an increasing pressure from the veterinary and medical bodies to revisit its use for treating and preventing diseases in dairy cows such as acute puerperal metritis (APM). The presence of Escherichia coli (E. coli) in the uterus of dairy cows at day 1 postpartum was shown to increase by 3 the risk of subsequently developing APM. Unfortunately, there are no data describing the efficacy of ceftiofur treatment to prevent APM in the subset of cows with E. coli in their uterus at day 1 postpartum. Therefore, the objective of the study was to quantify the efficacy of ceftiofur treatment in dairy cows with E. coli in their uterus at day 1 postpartum to prevent APM.

Identification of Escherichia coli and Trueperella pyogenes isolated from the uterus of dairy cows using routine bacteriological testing and Fourier transform infrared spectroscopy.

BackgroundUterine disorders are common postpartum diseases in dairy cows. In practice, uterine treatment is often based on systemic or locally applied antimicrobials with no previous identification of pathogens. Accurate on-farm diagnostics are not available, and routine testing is time-consuming and cost intensive. An accurate method that could simplify the identification of uterine pathogenic bacteria and improve pathogen-specific treatments could be an important advance to practitioners. The objective of the present study was to evaluate whether a database built with uterine bacteria from European dairy cows could be used to identify bacteria from Argentinean cows by Fourier transformed infrared (FTIR) spectroscopy. Uterine samples from 64 multiparous dairy cows with different types of vaginal discharge (VD) were collected between 5 and 60 days postpartum, analyzed by routine bacteriological testing methods and then re-evaluated by FTIR spectroscopy (n = 27).ResultsFTIR spectroscopy identified Escherichia coli in 12 out of 14 samples and Trueperella pyogenes in 8 out of 10 samples. The agreement between the two methods was good with a Kappa coefficient of 0.73. In addition, the likelihood for bacterial growth of common uterine pathogens such as E. coli and T. pyogenes tended to increase with VD score. The odds for a positive result to E. coli or T. pyogenes was 1.88 times higher in cows with fetid VD than in herdmates with clear normal VD.ConclusionsWe conclude that the presence of E. coli and T. pyogenes in uterine samples from Argentinean dairy cows can be detected with FTIR with the use of a database built with uterine bacteria from European dairy cows. Future studies are needed to determine if FTIR can be used as an alternative to routine bacteriological testing methods.

Presence of superantigen genes and antimicrobial resistance in Staphylococcus isolates obtained from the uteri of dairy cows with clinical endometritis

Clinical endometritis is an important disease of dairy cattle and results in decreased reproductive performance. This disease is caused by contamination of the uterus with a broad spectrum of microorganisms...

Genomic and phenotypic characterization of Escherichia coli isolates recovered from the uterus of puerperal dairy cows

The role of Escherichia coli in the pathogenesis of the puerperal uterine infection of the cow is largely unknown. It is proposed that E. coli favors the persistence of Arcanobacterium pyogenes and gram-negative bacteria that are pivotal to the establishment of the infection. Here, we report the genomic and phenotypic characteristics of 72 E. coli isolates recovered from the uterus of dairy cows with normal puerperium (n=12; 35 isolates) or clinical metritis (n=18; 37 isolates), in an attempt to identify characteristics that are related to the establishment of uterine infection. We evaluated DNA fingerprints generated by repetitive element sequence-based PCR, phylogenetic grouping, the presence of 15 virulence factor genes, in vitro biofilm formation and its relationship to curli fimbriae expression, and cellulose production. We found a wide genetic diversity (40 clonal types), including types common to normal puerperium and clinical metritis cows (n=6), as well as types specific to normal puerperium (n=14) or clinical metritis (n=20) cows. Isolates were assigned to phylogenetic groups B1 (58%), A (31%), and D (11%). Only 4 virulence factor genes were detected (hlyE, hlyA, iuc, and eaeA). In vitro biofilm formation was significantly affected by culture medium and incubation temperature. Curli fimbriae expression and cellulose production, although related to biofilm formation, were not required for it. None of the evaluated E. coli characteristics were significantly related to the establishment of the uterine infection. In conclusion, data presented in this paper indicate that E. coli isolates recovered from the uterus of puerperal cows present a wide genetic diversity, do not belong to a known pathogenic group, and have a low potential of virulence and persistence. This corroborates the putative role of the bacterium in the pathogenesis of the puerperal uterine infection of the cow.

Genomic characterization of Arcanobacterium pyogenes isolates recovered from the uterus of dairy cows with normal puerperium or clinical metritis

Arcanobacterium pyogenes is considered to be the most relevant bacterium involved in the establishment of puerperal uterine infection in cattle due to its persistence in utero, resistance to treatment and synergic action with Gram negative anaerobes. Once the infection is established, A. pyogenes is responsible for the persistence of the infection. The objective of this study was to characterize A. pyogenes field isolates recovered from the uterus of cows with either normal puerperium or clinical metritis, in an attempt to identify factors that might be associated with the establishment and persistence of the disease. This characterization was based on BOX-PCR typing and on screening of eight virulence factor genes ( plo, nanP, nanH, cbpA, fimA, fimC, fimE, fimG) by conventional PCR. Finally, a relationship between clonal types, virulence factors and presence of disease was investigated. A. pyogenes clonal types identified from isolates recovered from the uterus of postpartum dairy cows differed among herds. Although some clonal types were strictly associated with the development of clinical metritis, others were identified from isolates recovered from normal puerperium and clinical metritis cows. Moreover, the presence of the eight virulence factor genes was not related with the ability to induce clinical metritis, suggesting that the type of A. pyogenes may not be a determinant factor in the development of the disease. We suggest that host intrinsic factors, the synergism between A. pyogenes and other bacteria and the differential gene expression of virulence factor genes may play a more relevant role in the establishment of puerperal uterine infections.

Gross abnormalities, bacteriology and histological lesions of uteri of dairy cows failing to conceive or maintain pregnancy

AIM: To describe the gross pathology, bacteriology and histopathology of the reproductive tracts of dairy cows that failed to conceive or maintain pregnancy. METHODS: The reproductive tracts of 105 cows that were not pregnant at the end of the seasonal breeding programme were retrieved following slaughter. The tracts were examined grossly, both horns of the uterine lumen were swabbed for bacteriology, and tissue was collected from each horn and the body of the uterus for histopathology. Grossly enlarged uteri and tracts containing purulent vaginal content were excluded. Histopathology was performed on three sections of uterine tissue from each of three cows in which no gross pathological changes were detected and from which no bacteria were isolated, from three cows in which no gross pathology was detected but from which bacteria were isolated, and from three cows in which gross pathological changes were detected but from which no bacteria were isolated. RESULTS: Thirty-six (34%) cows had one or more gross lesions which involved the ovary, uterine tube, uterus or vagina. Bacteria were isolated from the uteri of 22 (21%) cows. Isolates included Arcanobacterium pyogenes (n=1), Escherichia coli (n=1), Fusobacterium spp (n=1), Haemophilus somnus (n=5), Streptococcus acidominimus (n=12), S. bovis (n=2), S. uberis (n=1) and S. salivarious (n=1). In only five cows were both gross pathology and bacteria detected. There was no relationship between the isolation of bacteria and the diagnosis of gross pathology of the uterus. There were no differences in the degree of histopathological changes in the uteri from the three groups of cows examined, and lesions present were minor. CONCLUSIONS: Gross pathological changes and intra-uterine bacteria were found in 34% and 20% of cows, respectively, but the correlation between the two was poor. Histopathological changes were unremarkable, suggesting the bacteriological findings were coincidental, that causative agents of infertility were not present at the time of examination, or that unrelated causes such as nutritional anoestrus may have been responsible for the failure of some cows to conceive.

Isolation and antimicrobial susceptibility of obligate anaerobic bacteria recovered from the uteri of dairy cows with retained fetal membranes and postparturient endometritis.

The uteri of 77 postparturient dairy cows were sampled. Samples were cultured aerobically and anaerobically, and the nature of bacterial growth was identified. A mixed aerobic and anaerobic bacterial infection was found in 55% of the samples. Actinomyces pyogenes was the predominant aerobic species; it was found in 70% of the samples, whereas Bacteroides melaninogenicus was the most frequent anaerobic species isolated. Altogether, 16 species belonging to the genus Bacteroides were identified with variable frequencies. It appears that more than one Bacteroides species colonizes the uterus of a given cow postpartum. The minimal inhibitory concentrations (MICs) of clindamycin, metronidazole, tetracycline and ciprofloxacin for 83 anaerobic isolates were determined. All isolates were susceptible to clindamycin (MIC90 of 0.064 microgram/ml) and all but two to metronidazole. Susceptibility to ciprofloxacin was variable, with a bimodal distribution of MIC values. The MIC of tetracycline for 90% of the isolates was > 256 micrograms/ml.

Isolation and antimicrobial susceptibility of [formula omitted] recovered from the uterus of dairy cows with retained fetal membranes and post parturient endometritis

The minimal inhibitory concentrations (MICs) of 14 antimicrobial agents for Actinomyces pyogenes isolates recovered from the uterus of cows with retained fetal membranes (RFM) and post-parturient endometritis (PPE) were determined. Cows with RFM received a course of intrauterine tetracycline therapy, whereas some cows with PPE received a similar treatment and others did not. Samples of uterine exudate were aseptically collected at least 10 d after the last treatment. A profuse growth of A. pyogenes was observed on blood agar plates inoculated with uterine exudate from 72% of the 54 cows sampled. Differences were not observed in the type and characteristics of bacterial growth in the exudate of cows with PPE which were sampled before and after antibiotic treatment. The MIC of penicillin G, amoxycillin, cephalothin lincomycin and tylosin for 90% of the isolates was <0.5 μg/ml; the corresponding value for streptomycin, oxytetracycline (OTC), chloramphenicol, norfloxacin and sulfadiazine/trimethoprim was >10.0 μg/ml. The MIC of OTC for 90% of the A. pyogenes isolates was >100 μg/ml. The susceptibility of 14 A. pyogenes isolates of bovine udder origin and 3 A. pyogenes ATCC isolates from the bovine udder and abscesses in sheep and swine to antibiotics was determined, for comparative purposes. Most of the isolates recovered from the uterus were resistant to OTC and sulpha/TMP whereas this was not the case for isolates recovered from other sources.

Spatial distribution of progesterone in bovine uterus in relation to corpus luteum location and function

Distribution of progesterone was determined in the uterus of dairy cows at different luteal stages of the estrous cycle. Tissue samples were excised at different and spatially distinct portions of the uterine horn (proximal, medial, distal) and body of the uterus. A significant (P< 0.05) interaction was observed between uterine horn and CL progesterone level groups. Concentrations of progesterone were higher in tissues of the uterine horn ipsilateral to the CL vs the body or contralateral horn but only from cows with high CL progesterone content. Progesterone concentrations were significantly higher (P<0. 05) in the proximal, medial and distal parts of the uterine horn ipsilateral to the corpus luteum than in any part of the contralateral horn and body of the uterus with high CL progesterone group. There was no significant interaction between uterine horn and sites. Concentrations did not differ among sites within either horn, regardless of CL activity. A positive relationship existed between uterine progesterone levels (means of all sites) in the ipsilateral (r=0. 75) and contralateral (r=0. 67) horns and CL progesterone levels. No apparent correlation was found between uterine tissue concentrations and ovarian follicular fluid or granulosa tissue concentrations. The results indicated that uterine tissue progesterone levels depend on CL activity (content) and that localized uterine increases are due to location (horn ipsilateral to an active CL).

The role of non-specific uterine infection in the infertility of clinically normal repeat-breeder cows

The role of non-specific bacteria in the infertility of clinically normal repeat-breeder cows is reviewed, and data are presented from a recent study in which bacteriological and histological samples were taken from the uteri of dairy cows at intervals during the non-gravid period. The evidence does not support the view that endometritis is an important cause of this infertility. In the absence of accurate counts of the bacteria present in the uterus at the time of service and during the subsequent oestrous cycle, it is impossible to determine whether pathogenic bacteria cause the infertility by injuring the gametes or the developing conceptus. A comprehensive quantitative study is needed of the microbiology of the uterus of the repeatbreeder cow at service and during the subsequent oestrous cycle.

Bacteriologic studies of bovine genital tracts: The use of dye as an indicator of postmortem contamination of the uterus by vaginal fluids

Non-specific bacteria were more frequently isolated from uteri of culled dairy cattle than beef cows (P <.01). In some studies, food dye was placed in the vagina just prior to slaughter as an indicator of postmortem contamination of the uterus by vaginal fluids and bacteria. The dye was more often found in the uteri of dairy cows than beef cows (P <0.05). Bacteria could always be isolated from uteri totally contaminated by the dye. Bacteria were more frequently isolated from uncontaminated areas of uteri that were partially contaminated by the dye, as compared to uteri in which no dye was found (P <.01). It is hypothesized that an enlarged cervical canal may allow dye and also bacteria to transverse the cervical canal into the uterus during the slaughtering process. It is further hypothesized that an enlarged cervical canal may more readily permit nonspecific bacteria of the vagina to ascend into the uterus in the live animal and cause reproductive failure.

Non-specific uterine infection and bovine fertility. II. Infection patterns and endometritis before and after service.

Abstract Bacteriological and histopathological studies of samples obtained from the uteri of dairy cows during the 14 days preceding first service and the 10 days immediately after the first and subsequent services did not indicate that the presence of non-specific infection or endometritis around the time of service was an important cause of infertility in cows which were free from clinically detectable uterine disease.