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Uterine epithelial cells (UEC) and migrated polymorphonuclear cells (PMN) play important roles in the uterine defence against microbial infection. The aims of the present study were to investigate i) whether undiluted uterine secretions modulate the expression of genes associated with the innate immune response in UEC and PMN in vitro, ii) whether these changes differ between the two cell populations and iii) whether uterine secretions from cows with subclinical endometritis produce a different response to those from unaffected cows. Therefore, undiluted uterine secretions, cytobrush and biopsy samples were collected from bovine uteri at a local abattoir. All cows had calved at least 3 months prior to sample collection. Subclinical endometritis was diagnosed by cytology (≥5% polymorphonuclear neutrophils) and histology. The uteri were thereby retrospectively categorised as endometritis-positive (E-pos; n = 14), if either the cytology or the histology results were positive, or endometritis-negative (E-neg; n = 17), if both diagnostics were negative. Cultured UEC responded to secretions from E-pos and E-neg cows with an increased gene expression of CXC ligand (CXCL) 8 and interleukin (IL) 6 compared to incubation with control medium alone. PMN expressed significantly higher mRNA levels of CXCL5, CXCL8 and IL1B in response to supernatant from UEC incubated with secretions from both groups (E-pos and E-neg) compared to those incubated with control medium alone. Gene expression of IL10 in uterine epithelial cells remained comparable to the control in cells exposed to E-pos secretions and was 3.6 times lower in those exposed to E-neg secretions. These results demonstrate that the expression of genes associated with the innate immune response in UEC and indirectly also PMN is affected by uterine secretions in vitro. Depending on the target gene, these changes differ between the two cell populations. UEC exposed to uterine secretions from cows without subclinical endometritis produce lower levels of IL10 compared to those exposed to secretions from affected cows or control medium alone. Therefore, the model established in this study can be used as a valuable tool to further understand the contributions of the two cell populations to the coordinated immune response in the uterus.

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Tumor infiltrating lymphocytes (TILs) serve as prognostic biomarker in human breast cancer. Rabbits have the potential to act as animal model for human breast cancer, and close similarities exist between the rabbit and human immune system. The aim of this study is to characterize TILs in pet rabbit mammary carcinomas and to statistically correlate results with histological and immunohistochemical tumor characteristics. Microscopic evaluation of TILs was performed in hematoxylin and eosin stained sections of 107 rabbit mammary carcinomas according to international guidelines for human breast cancer. Data on histological features of malignancy, estrogen and progesterone receptor status and calponin expression were obtained from the data base. This study revealed a statistical association between stromal TILs in the central tumor (CT) and infiltrative margin. Higher maximal percentages of stromal TILs at the CT were statistically correlated with decreased mitotic count and lower tumor grade. An increased number of calponin positive tumor cells was statistically associated with a lower mitotic count and a higher percentage of stromal TILs. Results suggest that higher percentages of stromal TILs are useful biomarkers that may point toward a favorable prognosis in rabbit mammary carcinomas and support the concept of the use of rabbits for translational research.

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The effects of standard uterine body and hysteroscopic insemination on endometrial health were investigated. For this purpose, 33 mares were assigned to five different protocols: control (no insemination; n = 7), sham AI (sham uterine body insemination; n = 6), sham HysAI (sham hysteroscopic insemination; n = 7), standard AI (standard uterine body insemination, 300 × 106 progressively motile sperms (PMS); n = 7) and HysAI (hysteroscopic insemination, 100 × 106 PMS; n = 6). Sampling included uterine swabbing for microbiological examination, cytology for determination of polymorphonuclear neutrophils (PMNs) in the uterus, and endometrial biopsy collection for histology and characterization of endometrial immune cells on day 18 after ovulation (B1) as well as 8-10 hours (B2, day 20) and 72 hours after insemination (B3, day 23). Microbial contamination increased throughout the experiment in the sham insemination groups. Significant effects (P < .05) over time were detected for PMNs (cytology: sham HysAI, standard AI, and HysAI; histology: standard AI and HysAI), macrophages (immunohistochemistry: standard AI and HysAI) and T cells (immunohistochemistry: standard AI), showing an increase at B2 and a subsequent decrease toward baseline levels at B3. At B2, significant differences (P < .05) existed for PMNs (mean ± SEM) between control (1.3 ± 1.9%) and sham AI (2.2 ± 2.7%) versus standard AI (12.2 ± 4.7%) and for macrophages between control (4.1 ± 3.5%) and sham AI (2.5 ± 1.3%) versus standard AI (25.4 ± 15.8%). Thus, the cellular immune response of the endometrium depends on sperm deposition in the uterus and does not differ between hysteroscopic and standard uterine body insemination.

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Mares are seasonally polyestric. The breeding season in spring and summer and the winter anestrus are flanked by transitional periods. Endometrial diseases are a frequent cause of subfertility and have an economic impact on the horse breeding industry. They include different forms of endometrosis, endometritis, glandular maldifferentiation, and angiosis. Except for suppurative endometritis, these are subclinical and can only be diagnosed by the microscopic examination of an endometrial biopsy. Endometrosis is characterized by periglandular fibrosis and nonsuppurative endometritis by stromal infiltration with lymphocytes and plasma cells. The pathogenesis of endometrosis and nonsuppurative endometritis is still undetermined. Some mares are predisposed to persistent endometritis; this has likely a multifactorial etiology. Glandular differentiation has to be interpreted under consideration of the season. The presence of endometrial diseases is associated with alterations in the expression of several intra- and extracellular molecular markers. Some of them may have potential to be used as diagnostic biomarkers for equine endometrial health and disease. The aim of this review is to provide an overview on pathomorphological findings of equine endometrial diseases, to outline data on analyses of cellular and molecular mechanisms, and to discuss the impact of these data on reproduction and treatment.

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Subclinical endometritis (SE) in cattle is defined as clinically unapparent inflammation of the endometrium. It is reported to impair fertility in affected cows and causes economic loss within the dairy industry. A gold standard for diagnosis of SE has not been set. Uterine cytology and histopathology are both applied, but low agreement between these methods has been described. The objective of the present study was to assess the capability of uterine secretions (US) as a new medium for diagnosis of SE. A novel sampling tool was applied to retrieve US as well as cytological, histological and bacteriological samples of the endometrium after a singular passage through the cervix in 108 dairy cows (43-62 days post-partum [dpp]). To assess the quality of the US samples, a proteome analysis of samples from five healthy donors was performed, demonstrating that in vivo sampling of US was feasible and generated samples suitable for diagnostic purposes. Diagnosis of SE was realized by the combination of clinical, cytological, and histopathological findings. Quantitative analysis of pro- and anti-inflammatory cytokines (interleukin (IL)1B, IL6, IL8, IL17A, IL10) in US was conducted using AlphaLISA-technology. RNAlater-fixed endometrial biopsies were used for gene expression analysis of the cytokines IL1B, IL6, IL8, IL10 and tumor necrosis factor alpha (TNFα) as well as the prostaglandin-endoperoxide synthase 2 (PTGS2) and the antimicrobial peptide S100A9 by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR). Cows were assigned to groups according to their uterine health status. A large group of animals (n = 83) displayed no signs of endometritis (E.NEG). Cytological and histopathological examination revealed low agreement; hence, animals with SE were differentiated into SE(cyto) and SE(histo) groups (n = 7 and n = 13, respectively). One animal in group SE(cyto + histo) as well as four animals with signs of clinical endometritis (CE) were excluded from further analysis. SE(cyto) showed significantly higher median concentrations of IL1B, IL8 and IL17A in US as well as a significantly higher median expression of IL1B, IL8 and IL10 in endometrial biopsies compared to E.NEG. No significant differences were found for IL6 and IL10 in US and IL6, TNFα, PTGS2 and S100A9 in endometrial tissue between these groups. SE(histo) presented no differences concerning the analyzed parameters compared to E.NEG. In conclusion, a method to sample US was successfully established in dairy cows. The cytokines IL1B, IL8 and IL17A are promising candidates in diagnosing cytological endometritis by US. Further assessment of US might contribute to a better understanding of the pathological mechanisms leading to chronic endometrial inflammation and to impaired fertility in affected cows.

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Most mammary tumors in pet rabbits are carcinomas; prognostic factors are unknown. The aim of this study on rabbit mammary carcinomas was to determine the expression of myoepithelial markers that have a prognostic relevance in human cancers. Mammary carcinomas (n = 119) of female or female-spayed pet rabbits were immunostained for cytokeratin AE1/AE3, vimentin, smooth muscle actin (SMA), and calponin; and percentages of non-neoplastic myoepithelial cells (ME cells) and calponin-positive neoplastic cells were determined. Using statistical analysis, data were correlated with the age of the rabbits and histological tumor characteristics. All carcinomas contained retained spindle-shaped ME, while 115 also contained hypertrophic ME (HME). A statistically significant relationship existed between a higher age and an increase in HME. In 111 carcinomas (93%), tumor cells expressed calponin. There was a significant correlation between higher percentages of calponin-positive tumor cells and a lower mitotic count, an increased percentage of tubular growth, and a lower grading score, respectively. Data suggest that pet rabbit mammary carcinomas develop from progression of in situ cancer and that the extent of calponin expression in tumor cells influences their biological behavior. These results provide the basis for a long-term follow-up on the prognostic significance of calponin expression in mammary cancer cells.

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The aim of this review is to raise awareness for mammary tumors in rabbits and to report progress in related research. Currently, a standardized tumor classification for rabbits is not available, prognostic factors are unknown and the only treatment option is surgical excision. Studies showed that affected rabbits have a wide age range and are nearly exclusively female or female spayed. Most mammary tumors are carcinomas. These may occur together with non-neoplastic or benign mammary lesions. Frequent microscopic findings are lipid droplets in tumor cells, secretory activity and microscopic heterogeneity. Since carcinomas are often negative for estrogen and progesterone receptors (ER-α/PR), modulation of receptor function will unlikely be beneficial for most rabbits. ER-α and PR status may have prognostic significance, since ER-α- or PR-negative tumors have significantly higher mitotic rates than ER-α- or PR-positive tumors. The frequent secretory activity of rabbit mammary tumors may suggest an influence of prolactin on tumorigenesis. Available data contribute to comparative pathology and are the basis for future molecular studies into the identification of additional prognostic factors and novel therapeutic options. They will also reveal the suitability of the rabbit as a model for certain types of breast cancer in women.

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Equine endometrosis, a frequent cause of subfertility, is characterized by periglandular fibrosis, and no treatment exists. Endometrial biopsies not only contain diseased glands, but also contain healthy glands and stroma. Myoepithelial (ME) and myofibroblastic (MF) markers are calponin, smooth muscle actin (SMA), desmin and glial fibrillary acidic protein (GFAP). Epithelial vimentin expression indicates epithelial to mesenchymal transition (EMT). The aim of this immunohistochemical study was to investigate whether biopsies with endometrosis express MF and ME markers and vimentin. Compared to healthy areas, significantly higher percentages of endometrotic glands were lined by calponin- and vimentin-positive epithelial cells, whereas periglandular fibrosis contained significantly higher percentages of stromal cells positive for vimentin, desmin and SMA and significantly less calponin-positive stromal cells. The rare GFAP expression was restricted to endometrotic glands. Of these, the most frequent features of endometrotic glands were higher percentages of SMA- and vimentin-positive stromal cells and the prominent epithelial calponin staining that occurred in 100%, 93% and 95% of examined biopsies. Results indicate plasticity of equine endometrial epithelial and stromal cells. Particularly, endometrotic glands show evidence for ME differentiation and EMT. The different expression of MF markers between stromal cells from healthy and endometrotic areas suggests functional differences. The characteristic changes in the expression of SMA, vimentin and calponin between endometrotic glands and healthy areas can be helpful to confirm early stages of endometrosis. The characterization of cellular differentiation may help to decipher the pathogenesis of endometrosis and could lead to therapeutic strategies.

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In mares, placental diseases are a common cause of pregnancy failure and they can have an economic impact on the horse breeding industry. To our knowledge no published data on TLR expression in the equine placenta exist. This study examined the expression of TLR 2, 4 and 6 as transcript and protein in the placenta (chorioallantois) of 14 foals born alive. By PCR, all examined placental samples contained TLR 2, 4 and 6 transcripts. Using immunohistochemistry, trophoblasts and allantoic epithelium were immunopositive for TLR 2, 4 and 6 in all placental samples. The majority of placental samples contained TLR 4 and 6 positive stromal cells and vascular smooth muscle cells. Since these results confirm the expression of TLR 2, 4 and 6 in different cell populations of the equine placenta, they are the basis for studies into the pathogenesis of TLR-associated placental diseases in mares.

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In breast cancer of women, the estrogen receptor-α (ERα) and progesterone receptor (PR) status has prognostic and therapeutic significance. The aim of this study was (1) to characterize by immunohistochemistry the expression of ERα and PR in nonneoplastic and neoplastic mammary gland tissue of pet rabbits and (2) to correlate the ERα/PR status and histological features. All 124 rabbits included in this study had a mammary tumor; in addition, 2 rabbits had lobular hyperplasia and 25 had multiple cysts. Of the 124 neoplasms, 119 (96%) were carcinoma, 2 (2%) were carcinoma in situ, and 3 (2%) were adenoma. ERα or PR or both were detected in 2 of 2 carcinomas in situ, 3 of 3 adenomas, 19 of 25 cysts, and 2 of 2 lesions of lobular hyperplasia. Most carcinomas (75/119, 63%) were negative for both ERα and PR; 22 of 119 carcinomas (18%) were double-immunopositive. The ERα and PR expression was not influenced by histotype or histological tumor grade. In carcinomas, there was a statistically significant correlation between increased mitotic count and reduced expression of ERα and PR, and the mitotic count was higher in double-immunonegative carcinomas (75/119). The findings suggest that in rabbit mammary carcinomas, proliferative activity is mainly influenced by factors other than estrogen and progesterone and provides the basis for future investigations into the prognostic significance of the ERα and PR status of mammary tumors.

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The enzyme indoleamine 2,3-dioxygenase 1 (IDO1) acts immunomodulatory and restricts bacterial growth. In the uterus of women and mice, it likely contributes to tissue homeostasis and disease pathogenesis. Pregnancy failure in mares is often caused by endometritis and endometrosis. The pathogenesis of nonsuppurative endometritis and endometrosis is still uncertain. To the authors' knowledge, no information on IDO1 expression in the equine endometrium is published. Aim of this study was to examine the presence of IDO1 as transcripts and proteins in the healthy and diseased endometrium of 25 mares and to determine its cellular expression. By PCR, IDO1 transcripts were detected in healthy (3 mares) and diseased endometria (22 mares). Western blot on 15 samples showed the concurrent presence of IDO1 proteins. Immunohistochemistry revealed its expression in macrophages and epithelial cells. Endometria of 21 mares showed an intense staining of glandular epithelia, whereas glands of the remaining 4 mares were negative or contained only few positive cells. Tissue samples of all mares showed a minimal to mild IDO1 expression in the surface epithelium and glandular ducts. Quantification of immunohistochemistry on biopsies of 6 mares collected at different stages of the same endometrial cycle indicated that the IDO1 expression is not influenced by the endometrial cycle. This study confirmed IDO1 expression also in the equine endometrium and suggests an immunomodulatory role of uterine macrophages and epithelial cells. A markedly reduced glandular IDO1 expression as detected in 4 mares may be associated with alterations of uterine immune defenses.

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In order to evaluate the influence of deoxynivalenol (DON) on histomorphological and immunohistochemical parameters in the development of porcine fetuses, five pregnant sows were fed a control diet (0.15 mg DON/kg diet) and seven sows a contaminated diet (4.42 mg DON/kg diet) between days 35 and 70 of gestation. On day 70, fetuses were delivered by caesarean section and sows and fetuses were euthanized. Tissue samples of three fetuses from each sow were collected, fixed in formalin, and processed routinely for light microscopy and immunohistochemistry. At necropsy, no macroscopic lesions were observed in any organ of the fetuses. Histomorphological, immunohistochemical, and morphometrical parameters of the immune system, liver, and intestinal tract were examined. The following antibodies were used in the liver, spleen, lymph nodes, thymus, gut, and bone marrow to compare control- and DON-treated animals: (I) CD3 and CD79a (T and B lymphocytes differentiation); (II) myeloid/histiocyte antigen 387 (MAC) (identification of macrophages); (III) Ki-67 Antigen (Ki-67) (proliferation marker); (IV) p-p-38 mitogen-activated protein kinases (p-p38 MAPK) as well as caspase-3 (cas3) and caspase-9 (cas9) (enzymes of apoptosis cascade); (V) tumor necrosis factor-alpha (TNFα) (immune-related protein). The results of the study show that exposure of pregnant sows with DON between gestation days 35 and 70 causes no pathomorphologically or immunohistochemically detectable alterations in all fetal organs examined.

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Subfertility in mares is mainly caused by endometrial diseases. Alterations of Toll-like receptors (TLRs) are associated with endometrial disorders in women. This study investigated TLRs 2, 4 and 6 in the equine endometrium. Endometria of 21 mares were examined by histology, PCR and immunohistochemistry. Tissues from 2 mares were considered normal. The remaining showed endometritis, endometrosis and/or angiosclerosis. TLRs 2, 4 and 6 were expressed as transcripts and proteins in all endometria. Immunohistochemistry detected TLRs 2, 4 and 6 in mast cells, luminal and glandular epithelial cells, stromal cells, endothelia, vascular smooth muscle and/or inflammatory cells. Between examined endometria numbers of immunopositive epithelial cells varied considerably; TLRs were located in their cytoplasm and/or the nucleus. All other cell types displayed a cytoplasmic staining. Results indicate a complex and cell-type-specific modulation of TLRs 2, 4 and 6 in the equine endometrium. The lack of a detectable association between a particular disease and a distinct cellular expression may be explained by the often combined presence of several factors with a possible influence on TLRs. This study expands the basic knowledge on equine endometrial immunity and will assist to uncover if immunological alterations contribute to uterine diseases of mares.

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Equine endometritis eosinophilica (EE) is rarely described and its diagnostic criteria are not defined. The aim of this study was to characterize histological features of EE. A data base (1995-2013) was searched for biopsies with increased eosinophils. This study included all biopsies with this diagnosis and representative biopsies without this record. The definition of equine EE was based on criteria for EE in women and the results of the determination of physiological numbers of eosinophils within the equine endometrium. EE was diagnosed in 55 mares. Biopsies of 10 mares contained eosinophils exceeding the physiological range, but no EE; the diagnosis of eosinophilic infiltrates (EI) was applied. Those of the remaining mares (n = 126) displayed eosinophils within the physiological range (EWPR). An irregular glandular differentiation during the breeding season was detected in 25% of mares with EE, 33% of mares with EI and 24% of the mares with EWPR. Most mares with EE (93%), EI (90%) and EWPR (72%) showed endometrosis; it was high grade in 11% with EE and 7% with EWPR. Endometritis was diagnosed within 56% of mares with EE, 40% of mares with EI and 37% of those with EWPR. In mares with EE suppurative endometritis dominated (58%) and in those with EWPR non-suppurative endometritis (58%). This study indicates EE as a primary fertility reducing disease. Results suggest an association between eosinophilic infiltration and the presence of neutrophils. Further, they provide the basis for future studies into the pathogenesis, prognosis and therapy of EE.

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Endometrial epithelial cells form a luminal barrier and are exposed to pathogens and non-infectious antigens. Toll-like receptors (TLRs) mediate pathogen defenses and tissue homeostasis, but are also involved in the pathogenesis of inflammatory and fibrosing alterations. Endometrial diseases are important causes of subfertility in mares. The pathogenesis of some types of persistent inflammation and periglandular fibrosis (endometrosis) is unknown. The aim of this study was to compare by in situ and in vitro immunohistochemistry the expression of TLRs 2, 4 and 6 in equine endometrial epithelial cells. An epithelial immunostaining for TLRs 2, 4 and 6 was detected in 76%, 57% and 90% of tissue sections, respectively. Positive cells lined the luminal surface, glandular ducts, mid glands and/or basal glands. An immunoreaction for TLRs 2, 4 and 6 was observed in 100%, 33% and 94% of cell cultures, respectively. The immunosignal was located in the cytoplasm and/or nucleus of endometrial epithelial cells under in situ and in vitro conditions. Results indicate a complex regulation of the epithelial expression of TLR 2, 4 and 6 proteins. The examined cell culture has to be regarded as suitable in vitro model. This study provides the basis for comparative investigations into the impact of different stimuli on the cellular expression of TLRs 2, 4 and 6. These will assist to find out if TLRs are involved in the pathogenesis of endometrial diseases and may help to understand as to why some mares develop persistent endometritis.

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In this study, endometrosis and angiosclerosis in mares were studied. Endometrosis is a severe, progressive, and irreversible fibrotic condition that affects the endometrium, whereas angiosclerosis refers to thickening of vessel walls due to degenerative changes leading to reduced elasticity of the walls and lower perfusion. Histologic evaluations were performed on biopsies and compared with vascular features of the endometrial surface obtained via narrow-band imaging (NBI) hysteroscopy. First, it was determined if hysteroscopic evaluation of the endometrium using NBI resulted in a better visualization of the vascular pattern (i.e., vessel-versus-background contrast was increased) compared with using white light. This was found to be the case for examinations in vivo (n = 10), but not when using abattoir uteri (n = 3). In the second part of this study, it was determined if vascular densities and sizes as derived from NBI images could be used as indicators for the degree of degenerative changes of the equine endometrium and its vessels. Narrow-band imaging hysteroscopic evaluations were performed (n = 10), and endometrial biopsies (n = 32) were collected. Histologic specimens were evaluated for degree of endometrosis and angiosclerosis, and they were classified in Kenney categories. Narrow-band imaging images were analyzed for vascular pattern. Samples classified to Kenney category I, or without signs of vessel degeneration, had significantly higher vascular densities than samples from Kenney category IIa or with angiosclerosis. In conclusion, narrow-band imaging facilitates enhanced visualization of the vasculature of the equine endometrium during hysteroscopies, which has applications in detection of endometrosis and angiosclerosis.

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BACKGROUND: The increasing life span of Callithrix jacchus in combination with the occurrence of metabolic and age-dependent diseases requires improved health surveillance for this species. METHODS: The health status of 56 marmosets was studied using a non-invasive blood pressure (BP) device. Age-, weight-, and sex-dependent changes were analyzed. Four animals with striking BP findings had follow-up exams. RESULTS: Physiological and pathological BP values could be defined. BP positively correlated with age and weight, while no effect of sex could be found. Measurement time for female and older animals was shorter than for male and younger individuals. Further analysis of the suspicious patients revealed renal or hepatic diseases and cardiac alterations. CONCLUSION: The description of age and weight influences on BP delivers physiological and pathological values for common marmosets. This may contribute to the understanding of aging process and cardiology in this primate species.

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The aim of this study was to evaluate the uterine blood supply and endometrial vessel architecture, during the equine estrous cycle. Narrow Band Imaging (NBI) hysteroscopy was used for evaluating changes in the endometrial vasculature during the estrous cycle [six mares, d 0 (representing the day of ovulation), d 6 and 11 in four locations]. In addition, endometrial biopsy samples were used for immunodetection of markers for angiogenesis (Vascular Endothelial Growth Factor A, its receptor 2, as well as angiopoietin-2 and its receptor-tyrosine-kinase Tie2) during the estrous cycle (three mares, d 0, 5 and 10; one biopsy per mare). Detailed analysis of hysteroscopic images revealed an increase in the vascular density from estrus towards diestrus. In contrast, microscopic specimens prepared from biopsies revealed no evidence for changes in the endometrial vessel number during the estrous cycle. Studies on expression of angiogenesis markers indicated that cyclic changes in the endometrial vascular density observed by NBI-hysteroscopy were not due to formation of new vessels. It is concluded that vessels are involved in blood supply of a smaller area during diestrus, facilitating better distribution of nutrients during this phase.

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This study was conducted to evaluate the effects of induced endometritis on uterine blood flow in cows. Transrectal Doppler sonography was performed on uterine arteries of six cyclic cows before and for 4 days after inducing acute endometritis by intrauterine infusion of 720 mg of policresulen, and for 4 days of the following estrous cycle. Time-averaged maximum velocity (TAMV) increased (p < 0.001) and pulsatility index (PI) decreased (p < 0.0001) within 1 h of policresulen administration, and did not change (p > 0.05) in the next 4 days of the same cycle. TAMV and PI values in the subsequent cycle did not differ (p > 0.05) from the values measured before infusion and showed no changes (p > 0.05) within the cycle. Blood flow parameters were not related (p > 0.05) to plasma concentrations of progesterone and estrogen. All cows showed an acute endometritis determined by histopathological findings of biopsy samples taken 1 day after infusion and fibrotic endometrial alterations detected in the subsequent cycle. No relationships were observed between fibrotic changes of the endometrium and uterine blood flow during either cycle. In conclusion, acute inflammation is accompanied by a rise in uterine blood flow, but fibrotic alterations do not seem to be related to Doppler sonographic findings.

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Brucellae are facultative intracellular pathogens causing disease in a wide range of domestic and wild animals as well as in humans. Brucella (B.) microti is a recently recognized species and was isolated from common voles (Microtus arvalis), red foxes and soil in Austria and the Czech Republic. Its pathogenicity for livestock and its zoonotic potential has not been confirmed yet. In the present study 25 SPF chicken embryos were inoculated at day 11 of age with 1.6×10(3) and 1.6×10(5)B. microti by yolk sac and allantoic sac routes. Re-isolation of B. microti indicated rapid multiplication of bacteria (up to 1.7×10(12)CFU). B. microti provoked marked gross lesions, i.e. hemorrhages and necroses. All inoculated embryos were dead (100% mortality) in between 2nd and 4th day post inoculation. The predominant histopathological lesion was necroses in liver, kidneys, lungs, spleen, gastrointestinal tract, spinal meninges, yolk sac and chorioallantoic membrane. Immunohistochemical examination showed the presence of Brucella antigen in nearly all of these organs, with infection being mainly restricted to non-epithelial cells or tissues. This study provides the first results on the multiplication and pathogenicity of the mouse pathogenic B. microti in chicken embryos. These data suggest that, even though chicken are not mammals, they could provide a useful tool for understanding the pathogenesis of B. microti associated disease.

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