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Background While fungal infections of the bovine uterus are well-known diseases in pregnant cattle, very limited knowledge exists on the presence and significance of fungi in the uterus of non-pregnant cows. Presence of fungi in the uterine lumen of postpartum (pp) cows has been reported, but little attention has been paid to this as most studies of the bovine pp uterus have focused on bacteria. Case presentation Microscopy of uterine lavage cytology slides of three cows from one herd revealed the presence of numerous yeast-like organisms, which were located either free in the fluid or within macrophages. Two of the cows were around 30 days pp, while the third was 7 months pp. None of the cows had been treated with antibiotics. Culturing of the flush samples was unsuccessful, but Sanger sequencing of DNA extracted from an endometrial biopsy of one of the cows revealed the presence of Candida kefyr (Kluyveromyces marxianus). Fluorescence in situ hybridization examination of endometrial tissue sections of two cows using probes targeting 18S rRNA of the K. marxianus group was performed and revealed the presence of yeast cells on the endometrium. Histology was performed and demonstrated hyphal and non-hyphal yeast-like organisms on the surface of endometrium and in the crypts. Tissue invasion was restricted to the superficial part of the epithelium and although endometrial inflammation was present, this was mild and considered as not being caused by the fungi. One of the cows became pregnant and delivered a normal calf at term, while the two others were not bred. Conclusions Candida kefyr is commonly isolated from milk of cows with mastitis, but has not been reported in association with other diseases of cattle. The infection was present as a monoculture in all three cows, but the fungi had only colonized the uterine lumen and the endometrial surface. Only a mild non-suppurative endometrial inflammation was present, but within the uterine l

Pyometra is a common disease of cattle that causes infertility and thereby financial losses to the cattle industry. Bacteria involved in the development and progression of pyometra have been investigated by microbial culture but their tissue invading abilities, which is an important aspect of bacterial pathogenicity and development of lesions, have not been investigated. Bacterial invasion of the uterus and oviducts was studied in 21 cows diagnosed with pyometra at the time of slaughter by applying fluorescence in situ hybridization using probes targeting 16S ribosomal RNA of Fusobacterium necrophorum, Porphyromonas levii, Trueperella pyogenes and the overall bacterial domain Bacteria. Fusobacterium necrophorum and P. levii were found to invade the endometrium, especially if the endometrium was ulcerated, and penetrated deep into the lamina propria. These species co-localized within the tissue thus indicating a synergism. Trueperella pyogenes did not invade the uterine tissue. In addition to endometrial lesions, most cows with pyometra also had salpingitis but without significant bacterial invasion of the oviductal wall.

The aim of this study was to investigate bacterial invasiveness of the bovine endometrium during the postpartum period. Fluorescence in situ hybridization was applied to endometrial biopsies using probes for Fusobacterium necrophorum, Porphyromonas levii, Trueperella pyogenes, Escherichia coli and a probe for bacteria in general (the overall domain Bacteria) to determine their tissue localization. Holstein cows were sampled at three time points postpartum (T1: 4–12 days postpartum, T2: 24–32 days postpartum and T3: 46–54 days postpartum). At T1, cows were clinically scored as having a uterine infection based on presence of a brownish, fetid vaginal discharge or as normal if having normal lochia. An endometrial biopsy was taken from all cows at T1 (n = 57). Endometrial biopsies were taken from the same cows at T2 and T3 if allowed by the size of the cervical canal and if the cow had not been inseminated. Fifty and 39 biopsies were obtained at T2 and T3, respectively. The biopsies were evaluated for inflammation and for presence and localization of bacteria. When analyzed by the probe for the entire domain Bacteria, bacteria were found in most biopsies irrespectively of time (T1: 79.0%, T2: 82.0%, T3: 89.7%). Fusobacterium necrophorum and Porphyromonas levii were often present in the endometrium at T1 (61.1% and 47.8%, respectively), but the prevalence decreased significantly over time. Trueperella pyogenes and Escherichia coli were less prevalent at T1 (8.8% and 10.5%, respectively) and their prevalence also decreased significantly over time. Fusobacterium necrophorum and Porphyromonas levii were often co-localized intraepithelially or in the lamina propria. Trueperella pyogenes and Escherichia coli were located only on the endometrial surface. Due to the high prevalence of tissue invasiveness, these findings emphasize the importance of Fusobacterium necrophorum and Porphyromonas levii in postpartum uterine disease of cattle and indicate that tissue invasiveness is a

The bacteria present in the uterus during pyometra have previously been studied using bacteriological culturing. These studies identified Fusobacterium necrophorum and Trueperella pyogenes as the major contributors to the pathogenesis of pyometra. However, an increasing number of culture-independent studies have demonstrated that the bacterial diversity in most environments is underestimated in culture-based studies. Consequently, fastidious pyometra-associated pathogens may have been overlooked. Therefore, the primary purpose of this study was to investigate the diversity of bacteria in the uterus of cows with pyometra by using culture-independent 16S rRNA PCR combined with next generation sequencing. We investigated the microbial composition in the uterus of 21 cows with pyometra, which were obtained from a Danish slaughterhouse. Similar to the observations from the culture studies, Fusobacteriaceae, the family that F. necrophorum belongs to, was the operational taxonomic unit (OTU) observed in the largest quantities. By contrast, the Actinomycetaceae family, which includes T. pyogenes, constituted only 1% of the total number of reads. Thus we cannot confirm the previously reported role of species from this family in the pathogenesis of pyometra. Finally, we identified a large number of sequences representing three families of Gram-negative bacteria in the pyometra samples: Porphyromonadaceae, Mycoplasmataceae, and Pasteurellaceae. It is likely that these families comprise potential pathogenic species of a fastidious nature, which have been overlooked in previous studies. Our results increase the knowledge of the complexity of the pyometra microbiota and suggest that pathogens in addition to F. necrophorum may be involved in the pathogenesis of pyometra. (C) 2014 Elsevier B.V. All rights reserved.

Recent years’ advance in sequencing technology has resulted in extensive new knowledge of the microbial ecology of different environments. We used the technology to investigate the causality of endometritis, which is an inflammation in the inner lining of the uterus affecting up to 20% of dairy cows in Denmark post partum. Endometritis is linked to reduced reproductive performance, which is costly for the farmer and often leads to culling of the affected cows. With incomplete knowledge of the bacteria involved, treatment is performed without an option for choosing the best suited antimicrobial agent, which may lead to unnecessary antibiotic resistance development. Slaughterhouse samples were analysed in order to obtain information on the uterine microbiota from both cows with endometritis and seemingly healthy cows from a variety of herds. We sampled uteri from cows (n=50) from a slaughterhouse in Holstebro, Denmark. An incision was made into the right uterine horn and an endometrial biopsy was taken with a pair of sterilised scissors. The endometrial surface was sampled with a cotton-swab through the same incision. All samples were immediately put in RNAlater. The DNA was extracted with the Maxwell 16 LEV Blood kit (Promega), the 16S rRNA PCR was performed with primers targeting the V2 region, and the 454 next generation sequencing was performed by GATC. Previous results have shown that Proteobacteria and Tenericutes are the most important bacteria phyla in the uterus of healthy cows (Machado et al. 2012 and Galvão et al. 2012) while Escherichia coli, Trueperella Pyogenes, Prevotella species and Fusobacterium necrophorum have commonly been associated with endometritis (Sheldon 2006). Preliminary results indicate that we often find bacteria from phylum Actinobacteria in the healthy cows and often bacteria from phyla Bacteroidetes and Fusobacteria in cows with endometritis.

Hos mennesker med venstresidig endokarditis opstår der hyppigt neurologiske komplikationer. Patientgruppen der udvikler neurologiske komplikationer har en signifikant højere dødelighed end de patienter der ikke udvikler neurologiske komplikationer til venstresidig endokarditis. For at udbygge viden om biologiske processer bag og behandling af neurologiske komplikationer som følge af endokarditis, er der behov for at udføre forsøg på dyremodeller. I forbindelse med behovet for dyremodeller ønskes det undersøgt, om der hos svin er en sammenhæng mellem venstresidig valvulær endokarditis og udvikling af læsioner i hjernen. På baggrund af resultaterne foretages, en vurdering af om svin er egnede som dyremodel til humanforskning af hjernelæsioner som komplikation til endokarditis. Der blev til denne undersøgelse udvalgt 20 svin med venstresidig valvulær endokarditis og en kontrolgruppe på fem svin. Der blev udarbejdet 763 histologiske snit af hjernerne fra alle svin, og alle snit blev systematisk undersøgt i mikroskop. Der blev observeret seks forskellige typer læsioner i hjernerne fra svinene med endokarditis; Perivaskulære cuffings, encefalitis, leptomeningitis, vaskulitis, ødemer og ét infarkt. I alt blev der observeret 115 mikroskopiske læsioner i risikohjernerne. Fundene var fordelt mellem alle dele af hjernen (cerebrum, cerebellum og hjernestammen). De hyppigst observerede læsioner var perivaskulære cuffings og encefalitis. Konklusionen på undersøgelsen blev, at der er en sammenhæng mellem tilstedeværelse af venstresidig valvulær endokarditis hos svin og udviklingen af hjernelæsioner. Det blev ligeledes konkluderet at svin som udgangspunkt kan benyttes som dyremodel for encefalitis opstået som komplikation til venstresidig valvulær endokarditis, men at det først skal verificeres om der også opstår hjernelæsioner som komplikation til eksperimentelt induceret endokarditis hos svin. Det vurderes, at svin ikke kan benyttes som dyremodel for infarkter opstået som komplik, It is often seen that humans with endocarditis gets neurologic complications. The patients who contract these neurologic complications have an increased risk to die compared to patients who do not contract neurologic complications. Animal models are important tools to improve the scientific knowledge about the biological processes and to test new medicine and treatments. Regarding this need for using animal models, this study aims to prove whether there is a correlation between suffering from valvular endocarditis and developing cerebral lesions. On the basis of the findings, it will be evaluated whether swine can be useful as animal models for cerebral lesions secondary to valvular endocarditis in human medical research. Two groups of swine were used in this study, one group with 20 swine with valvular endocarditis and five swine without. 763 histological slides were made from the brains in both groups, and were thereafter evaluated systematic in a microscope. Six different types of lesions were found in the brains; - Perivascular cuffings - Encephalitis - Leptomeningitis - Vasculitis - Edema - Infarct All findings were microscopic in sizes and were found in all parts of the brain (cerebrum, cerebellum and brainstem). The conclusion was that there is a correlation between suffering from valvular endocarditis and developing cerebral lesions. Swine can be useful in human research in the study of encephalitis secondary to valvular endocarditis, if it is possible to verify development of encephalitis secondary to experimental induced endocarditis in swine. It is realized on basis of this study that swine are not suitable for use as animal models in study of infarcts secondary to valvular endocarditis. Besides this, it is in this study not found possible to estimate the possibility of brain lesions in swine with endocarditis, based on the presence of infarcts in the kidneys.