Aspects of transition cow metabolomics-Part II: Histomorphologic changes in the liver parenchyma throughout the transition period, in cows with different liver metabotypes and effects of a metaphylactic butaphosphan and cyanocobalamin treatment.

The aims of this study were to evaluate histopathologic changes during the transition period, describe the histopathological features of the metabotypes identified in Part I (Schären et al., 2021b), and investigate effects of a metaphylactic treatment with butaphosphan and cyanocobalamin (BCC) on the liver parenchyma. Eighty German Holstein cows (mean 305-d production: 10,957 kg, range: 6,480-15,193 kg; mean lactation number: 3.9, range: 2-9) from a commercial dairy farm in Saxony, Germany, were enrolled in a randomized, prospective, triple-blinded study. Two groups received a treatment with BCC (5 or 10 mL/100 kg of body weight 10% butaphosphan and 0.005% cyanocobalamin, Catosal, Bayer Animal Health, n = 20 each) and one group a placebo treatment (NaCl 0.9%, n = 40). Liver biopsy specimens were collected 14 d antepartum (AP) and 7, 28, and 42 d postpartum (PP), routinely processed for histologic examination, and stained with hematoxylin and eosin, Sudan III, periodic acid-Schiff, and picrosirius red stains. The sections were assessed for fat and glycogen content and degenerative, inflammatory, fibrotic, and proliferative changes. The statistical analysis included the effects of the sampling day, the lactation number, the treatment, and the metabotype (A = medium, B = minor, C = large alterations in the liver metabolome profile between AP and PP status). There was mild to moderate fat infiltration in the liver of 37% of cows in the last 2 wk AP, and moderate to severe fat infiltration in 66% of cows in the first days PP. The degree of fat infiltration increased from 2 wk AP until the end of the first week PP, and then decreased until the end of the study period, at which time about 25% of cows had moderate to severe fatty infiltration. Lipidosis was positively correlated with the severity of liver cell degeneration, and negatively correlated with the degree of glycogen deposits. Complete glycogen depletion of hepatocytes was not observed in cows, even in the presence of severe hepatic lipidosis. Moderate to severe lymphocytic hepatitis was seen in 39% of cows throughout the study period, and cows with lactation numbers 5 or greater had perisinusoidal fibrosis more often than younger cows. Severe fibrosis and cirrhosis of the liver did not occur. Metabotype B animals exhibited a higher chance of fatty infiltration, lower glycogen storage, and perisinusoidal fibrosis and for this metabotype positive correlations were calculated between increased fat deposition in the liver and marked glycogen depletion, and increased degenerative, inflammatory, fibrotic, and proliferative changes of hepatic tissue. For the treatment with BCC, no significant effect was observed. In summary, during the transition period, the liver of dairy cows is characterized by fat accumulation and glycogen depletion and histologic signs of hepatitis and hepatocyte degeneration. These histomorphologic changes were accentuated in animals exhibiting little alterations in their liver metabolome profile across the transition period (metabotype B) and support the assumption of a decreased grass silage quality as a causative factor.

Cytokeratin expression profiles of canine epithelial tissues.

Cytokeratins (CKs) are intermediate filaments of epithelial cells. In humans, different types of epithelia as well as their neoplasms show distinct CK expression profiles. The aim of this study was to establish a panel of CKs for the identification of specialized canine epithelia that can be integrated in a routine diagnostic setting. Immunohistochemistry was performed on 42 formalin-fixed paraffin-embedded (FFPE) canine unaltered tissues including all epithelial tissues by using an antibody panel detecting CKs 7, 8, 13, 14, 17, 19 and 20 and the pancytokeratin marker AE1/AE3. Using this antibody panel, a differentiation scheme for the identification of canine tissues was developed. This allowed the identification of 23 out of the 42 examined canine tissues and the distinction of 9 groups of specialized epithelia. The statistical validation revealed high variations in the immunoreactivity for CKs 7, 8, 14, 17 and 20 between the donor dogs. The antibody detecting CK 7 (OV-TL 12/13) showed a decrease in immunostaining after a fixation time of 3 and 4 days. To the best of the authors' knowledge this is the first study that characterizes all canine epithelial tissues for their expression of CKs 7, 8, 13, 14, 17, 19 and 20 and the pancytokeratin marker AE1/AE3. Results of this study are an important prerequisite for comparative histology and for the investigation into similarities/differences of the cytokeratin expression between normal and neoplastic epithelia. Since this study was performed on FFPE tissue, it can be included in the workflow of a routine diagnostic laboratory.

Effects of oxytocin and PGF2α on uterine contractility in cows with and without metritis-An in-vitro study.

The aim of this study was to investigate the effects of PGF2α and oxytocin in vitro on myometrial contractility in puerperal uteri. Thirteen puerperal uteri were removed and perfused after euthanasia of cows with (n=7) and without metritis (n=6). Measurement of uterine contractility was done using four piezoelectric crystals, which were implanted into the myometrium along the greater curvature of the uterine horn where fetal implantation occurred during the previous pregnancy. After 30min of equilibration, oxytocin (5 IU) or PGF2α (2.5mg Dinoprost) was administered randomly into both uterine arteries, and 30min later, the second administration of either oxytocin or PGF2α occurred. Treatment with oxytocin induced contractions in uteri with metritis and uteri without metritis (P<0.05). In uteri with metritis, greater uterine contractions occurred after stimulation with oxytocin than in uteri without metritis (P<0.05). Treatment with PGF2α did not (P>0.05) result in increased contractions in the uteri without metrtitis, however, induced an initial decrease in contractions followed by an increase (P<0.05) in contractions in uteri with metritis. Myometrial and endometrial gene expression of PGF2α (FPR) and oxytocin receptor (OTR) was greater (P<0.05) in uteri with metritis than in uteri without metritis. The results suggest that oxytocin, but not PGF2α, is an effective uterotonic drug in puerperal cows. Uteri in which metritis was diagnosed contracted more strongly after treatment with oxytocin than uteri in which metritis was not diagnosed. This effect was paralleled by greater gene expression of OTR as well as FPR in uteri with metritis compared with uteri in which metritis was not diagnosed.

Evaluation of the iVET® birth monitoring system in primiparous dairy heifers.

The objective was to validate the iVET® birth monitoring system and to determine if it reduced fetal death in primiparous dairy heifers over a 1-y interval. There were 359 pregnant heifers enrolled; 167 heifers in the iVET® group were monitored electronically and the remaining 192 (controls) were monitored visually for onset of Stage 2 labor, according to routine farm management. In addition, as a reference, all heifers were observed throughout the study by two independent investigators. Calves born dead or that died within 24 h after birth were defined as stillborn. The interval from appearance of the chorioallantoic sac to recognition of onset of calving in the control group averaged 21 min longer than the iVET® signal (p = 0.0001) and rate of fetal death was numerically lower in the iVET® group (8.9%) than in the control group (10.4%, p = 0.65). Interestingly, dystocia occurred more often in the iVET® group (58.3%) than in the control group (40.9%, p = 0.001). The iVET® system detected onset of Stage 2 labor earlier than conventional monitoring by farm staff. However, the device was lacking in several aspects and should be improved before its use in primiparous heifers can be recommended.

Seasonal or pathological findings? Morphofunctional characteristics of the equine endometrium during the autumn and spring transition.

The deep anoestrous phase in winter is part of the anovulatory season in mares and is bordered by the autumn and spring transitional periods (ATP/STP). To define an annual time span for effective prognostic biopsy sampling, the aim of this study was to provide a morphofunctional characterization of the endometrium during ATP and STP. To outline both transitional periods, endometrial specimens were taken in September, October and November (n = 76) as well as February, March and April (n = 184) with the requirement of a detailed clinical documentation. Tissue samples were examined histologically with special emphasis on the functional endometrial morphology. Additionally, an immunohistochemical evaluation was performed on selected specimens regarding the expression of oestrogen receptor α, progesterone receptor and Ki67-antigen. An absent to low endometrial activity was ascertained in more than 60% of all specimens from late October onwards, whereas a comparably lacking or low activity in STP was observed until early April. Approximately 30% (ATP) to 22% (STP) of all samples exhibited a predominantly "irregular" endometrial differentiation. During the transitional periods, the clinically evaluated ovarian status (transrectal palpation, transrectal ultrasonography and/or serum progesterone and oestrogen analyses) and the endometrial functional morphology were in accordance with approximately 70% of all cases. The expression of steroid hormone receptors and Ki67-antigen was generally low. Given that endometrial maldifferentiations were frequently found during ATP and STP, its occurence might display a characteristic and physiological feature of the transitional periods. Regarding the functional endometrial morphology, a diagnostic biopsy sampling should therefore be performed between late April and before September.

Establishment and characterization of a coculture system of equine endometrial epithelial and stromal cells.

To investigate the equine endometrium as close to the in vivo situation as possible, we established a coculture system for epithelial and stromal cells (ECs/SCs). ECs and SCs were isolated from nine endometrial tissue specimens. ECs obtained as glandular formations were cultivated on one side of the semipermeable membrane of a Millicell® insert. After 2 days, SCs (2 × 104 cells/membrane) were seeded onto the other side of the same membrane. During cocultivation, the low serum containing culture medium (Theuß et al., 2010) was supplemented with different concentrations and combinations of 17ß-estradiol (2.0-3.0 pg/ml medium) and progesterone (0.5-15.0 ng/ml medium). Once the cocultures formed continuous cell layers as determined by phase-contrast microscopy, the membranes were fixed and processed for light microscopical examination. Cytokeratin 19, steroid hormone receptors and the uterine proteins uteroglobin and calbindinD9k were detected using immunocytochemistry to determine the degree of culture purity and functional cellular differentiation. The culture purity of the EC layer averaged ≥95%. Uteroglobin and calbindinD9k were consistently expressed in ECs, while hormone receptors were predominantly absent in both cell populations. An explicit cytomorphological epithelial differentiation with formation of round-oval to polygonal cell forms was encountered in ≤50% of all ECs and independent of supplemented steroids. Based on the findings altogether, and despite the partly absent congruence to the in situ prerequisites, we established a standardized and reproducible coculture system, which offers a basic approach for studies of physiologic and pathophysiologic issues in the mare.

Prototheca species and Pithomyces chartarum as Causative Agents of Rhinitis and/or Sinusitis in Horses.

Pyogranulomatous rhinitis associated with an algal infection was diagnosed in a 25-year-old gelding and a 23-year-old mare had necrotizing sinusitis with intralesional algae and pigmented fungi. Algae were identified immunohistochemically in both cases as Prototheca spp. In the gelding, further characterization by polymerase chain reaction and sequencing revealed that the organism was Prototheca zopfii genotype 2. Fungi from the mare were identified as Pithomyces chartarum by molecular analysis. Prototheca species are achlorophyllous algae and P. chartarum represents a dematiaceous fungus; they are saprophytes and facultative pathogens. Prototheca spp. and P. chartarum should be considered as rare respiratory pathogens of horses.

The effect of puerperal uterine disease on histopathologic findings and mRNA expression of proinflammatory cytokines of the endometrium in dairy cows.

The aim of this study was to investigate the effect of puerperal uterine disease on histopathologic findings and gene expression of proinflammatory cytokines in the endometrium of postpuerperal dairy cows; 49 lactating Holstein-Friesian cows were divided into two groups, one without (UD-; n = 29) and one with uterine disease (UD+; n = 21), defined as retained fetal membranes and/or clinical metritis. General clinical examination, vaginoscopy, transrectal palpation, and transrectal B-mode sonography were conducted on days 8, 11, 18, and 25 and then every 10 days until Day 65 (Day 0 = day of calving). The first endometrial sampling (ES1; swab and biopsy) was done during estrus around Day 42 and the second endometrial sampling (ES2) during the estrus after synchronization (cloprostenol between days 55 and 60 and GnRH 2 days later). The prevalence of histopathologic evidence of endometritis, according to the categories used here, and positive bacteriologic cultures was not affected by group (P > 0.05), but cows with uterine disease had a higher prevalence of chronic purulent endometritis (ES1; P = 0.07) and angiosclerosis (ES2; P ≤ 0.05) than healthy cows. Endometrial gene expression of IL1α (ES2), IL1ß (ES2), and TNFα (ES1 and ES2) was higher (P ≤ 0.05) in the UD+ group than in the UD- group. In conclusion, puerperal uterine disease had an effect on histopathologic parameters and on gene expression of proinflammatory cytokines in the endometrium of postpuerperal cows, indicating impaired clearance of uterine inflammation in cows with puerperal uterine disease.

Prevalence of cervicitis in dairy cows and its effect on reproduction.

The objective of this study was to determine whether cervicitis in dairy cows is an independent disease or occurs concomitantly with inflammation of the uterus, and to clarify possible effects of cervicitis on reproductive performance. Dairy cows (n = 416) from 33 dairy farms were examined by rectal palpation and vaginoscopy between 42 and 50 days postpartum. Inclusion criteria for this study were absence of abnormal vaginal discharge and abnormalities of the uterus (fluctuation) at rectal palpation. Cervicitis was diagnosed when the second cervical fold was swollen and prolapsed with (C2) or without (C1) reddening. Cytobrush samples from the uterus (n = 370) and the cervix (n = 402) were collected, and the percentage of neutrophils in the uterus (PMNU) and the cervix as indicators of inflammation (threshold: ≥5%) was determined. In addition, endometrial biopsies for histology were collected, 300 of which were suitable for evaluation. Cervicitis (C1/C2) was diagnosed in 253 of 416 (60.8%) of cows. Of these, the prolapsed cervical mucosa was hyperemic (C2) in 29.1% of cases. Of 370 available uterine cytology samples, 221 cows had a clinical cervicitis; however, 170 (76.9%) had PMNU less than 5%. Of 300 uterine histologic examinations, 82 (27.3%) did not reveal any abnormalities; the remaining cows either had uterine inflammation and/or degenerative uterine changes such as endometriosis and angiosclerosis. Furthermore, of 300 biopsied animals, 184 revealed a cervicitis (C1/C2); however, 30.4% of these animals had no histopathologic uterine findings. For further analysis, only animals either without histopathologic findings and normal uterine cytology or with solely endometritis (defined as PMNU ≥ 5% and/or positive histopathology of the uterine tissue) were evaluated (n = 157). Of these, 95 cows had cervicitis. Unexpectedly, 63 of 95 (66.3%) cows had cervicitis without endometritis. With regard to reproductive performance, days to first service were not affected by cervicitis. Number of days open in animals with cervicitis but without endometritis tended to be lower than in cows with cervicitis plus endometritis (P = 0.092). Also, number of days open relative to percentage of neutrophils greater than 5% was lower when the cervical compared to the uterine mucosa was affected (P < 0.05). Total conception and pregnancy rates of animals 200 days into lactation decreased significantly in cows with severe cervical inflammation (C2). In conclusion, the results of this study suggested that cervicitis occurs independent of endometritis, and a higher degree of cervicitis is associated with poorer reproductive performance.

Effects of Intrauterine Devices in Mares: A Histomorphological and Immunohistochemical Evaluation of the Endometrium.

Oestrous suppression by intrauterine devices (IUDs) is caused by prolongation of luteal function, but the biological mechanism is unknown. The aim of the study was to investigate mechanisms which could explain the action of IUDs. Thirty mares were age-matched and either inseminated (AI, n = 15) or fitted with an IUD (IUD, n = 15) and subsequently divided into four groups: AI-P, pregnant (n = 8); AI-N, non-pregnant (n = 7); IUD-P, prolonged luteal phase (n = 7); and IUD-N, normal luteal phase (n = 8). The median ages were 5.5 and 7 years in AI-P and IUD-P groups and 14 and 11 years in AI-N and IUD-N groups, respectively. On Day 15 after ovulation, an endometrial biopsy was obtained to study histomorphological and immunohistochemical expression patterns of uterine proteins (uteroferrin, UF; uterocalin, UC; uteroglobin, UG), oestrogen and progesterone receptors (ER, PR), proliferation marker Ki-67 and content of inflammatory cells. Expression of UF was higher in IUD mares; the difference between pregnant and IUD-P mares was significant. Mares exhibiting a prolonged luteal phase (AI-P, IUD-P) showed only mild angiosclerosis and lower expression of both ER and PR than mares with a normal luteal phase (AI-N, IUD-N). No significant differences were detected in the numbers of inflammatory cells, with the exception of macrophages, which were more numerous in AI-P than AI-N mares. Although inflammatory cells were not detected in IUD mares, increased UF levels may indicate chronic inflammation. Young age and normality of the endometrial blood vessels may improve the efficacy of IUDs.

Effects of an anabolic steroid (Durateston) on testicular angiogenesis in peripubertal stallions.

The aim of this study was to determine the effect of the anabolic steroid testosterone on the testicular vascularization, angiogenesis and expression of angiogenic factors, and their receptors in testes of peripubertal stallions. Seven peripubertal stallions were treated with Durateston and castrated either 4 (treatment group 1 [TG1]) or 12 weeks (TG2) after the last injection. The castration of seven untreated control stallions (control group [CG]) took place within the same time. In the testicular specimens, volume density (VD), numerical density (ND), and area of vessels were determined morphometrically. Immunohistochemically, the expression of vascular endothelial growth factor (VEGF) and its receptor VEGF-R2; angiopoietin 2 (Ang2) and its receptor Tie2 as well as of transforming growth factor α (TGF-α) was investigated. Morphometrically, the VD of TG1 (P = 0.000) and TG2 (P = 0.001) vessels and the ND of arterioles and venules and capillaries were higher (TG1, TG2: P < 0.05), and the area of capillary cross sections was smaller (TG1, TG2: P < 0.05) than that in the CG. Compared to TG2 horses, TG1 animals showed a higher (P < 0.05) VD and ND of vascular structures and a smaller (P < 0.05) area of capillary cross sections. In numerous vascular structures, especially of TG1, the TGF-α and, to a less extent, the Ang2 and VEGF-R2 expression was significantly higher (P < 0.05) than that in the CG. Sertoli cells in TG2 were characterized by a significantly higher expression (P < 0.05) of VEGF-R2 than in the CG. In summary, the most and smallest vessels could be detected in the testes of TG1. Most likely this is explainable by the highest expression of some angiogenic factors (TGF-α, Ang2) and receptors (VEGF-R2) investigated. This expression behavior may be stimulated by testosterone. As a significant decrease of morphometric parameters could be detected in TG2 compared to TG1, the stimulatory effect of testosterone seems to be temporary.

Effect of suppression of postpartum ovulation on endometrial inflammation in dairy cows.

The objective of this study was to investigate the effect of time of first postpartum ovulation on endometrial inflammation in dairy cows with and without uterine disease during the early puerperal period. Transvaginal follicular puncture (FP) was carried out to suppress postpartum ovulation and formation of a CL until Day 42. Fifty-three lactating Holstein Friesian cows were divided into four groups on the basis of presence (UD+) or absence (UD-) of uterine disease, which was defined as retained fetal membranes and/or metritis, and whether FP had (FP+) or had not been (FP-) carried out. This resulted in the following groups: UD-FP- (n = 15), UD-FP+ (n = 13), UD+FP- (n = 13), and UD+FP+ (n = 12). Cloprostenol was given on Days 55 to 60 postpartum, and GnRH was administered 2 days later for synchronization of ovulation. In the FP- groups, endometrial swab and biopsy samples were collected during the second estrus (approximately Day 40) and during the estrus after synchronization. In the FP+ groups, the same samples were collected during the first estrus (approximately Day 49) and during the estrus after synchronization. The prevalence of positive bacteriologic cultures of the endometrium was not affected by FP (P > 0.05). Histologic signs of endometritis were more severe in UD+FP- cows at second sampling than in UD+FP+ cows (P ≤ 0.05). Endometrial expression of IL1α (in UD- after first sampling and in UD+ after second sampling) and IL1ß (in UD- and UD+ after first sampling) was higher (P ≤ 0.05) in FP- cows than in FP+ cows. Regardless of group, cows with histopathologic evidence of endometritis had higher expression (P ≤ 0.05) of IL1α, IL1ß, IL6, and TNFα than cows without endometritis. In conclusion, suppression of early ovulation by transvaginal FP enhances clearance of uterine inflammation in postpartum cows.

[Findings in uteri and ovaries from Eringer cows slaughtered due to fertility problems]. / Befunde von Uterus und Ovarien geschlacbteter Eringerkühe mit Fruchtbarkeitsproblemen.

Eringer cows are often slaughtered due to fertility problems which result from inflammatory and degenerative changes of the uterus or hormonal imbalances. Twenty-one genital tracts from Eringer cows suffering from fertility problems were collected in the abattoir. The purpose of the study was the macroscopic evaluation of the ovaries and the uterus followed by a histological and microbiological analysis of the uterus. Data from inseminations and calvings were provided by the Eringer breeding association and through the internet portal www.agate.ch. Median age of the cows was 6.9 years, number of calves per cow was 2.5 and median period between last calving and slaughter was 1.5 years. In 13 from 21 of the urogenital tracts examined, macroscopic abnormalities of the ovaries and/or histologic or microbiologic findings in the uterus could explain fertility-associated slaughter.

[Pathology of South American Camelids: a retrospective study of necropsies at the Institute of Veterinary Pathology, University of Leipzig, Germany]. / Sektionsbefunde von Neuweltkameliden. Eine retrospektive Analyse des Sektionsgutes des Leipziger Instituts für Veterinär-Pathologie.

OBJECTIVE: The number of South American Camelids (New World Camelids) housed in Germany has increased in the recent years. While these species were formerly kept solely in zoological gardens, ever more private and commercial livestock is being established. Compared to indigenous livestock animals, they bear some distinctive differences, particularly in terms of digestive tract anatomy and physiology. Therefore, it is of considerable interest for veterinarians working with South American Camelids to obtain knowledge about the distinguishing features of these animals and the typical diseases affecting them in Germany. MATERIAL AND METHODS: For this purpose, the necropsy reports, including the anamnestic data, and their diagnostic usefulness, from 1995 to 2012 were studied retrospectively. RESULTS: Du- ring this period, a total of 233 New World Camelids were examined (195 alpacas and 38 llamas). Anamnestic data of diagnostic usefulness regarding the cause of disease were only submitted in a limited number of cases, because most of the animals died without specific symptoms. The following were the most frequent pathological findings: enteritis (n = 91), gastritis (n = 76), cachexia (n = 73), pneumonia (n = 30), stomatitis (n = 27), azotaemia (n = 22) and anaemia (n = 9). An endoparasitosis occurred in 107 cases and was considered the predominant cause of enteritis. CONCLUSION: As with indigenous ruminants, llamas and alpacas primarily suffered from diseases of the digestive and respiratory tracts. Other organ systems were affected to a lesser extent. CLINICAL RELEVANCE: Even in cases with severe alterations in the affected organs, South American Camelids do not show or show too late diagnostically indicative clinical symptoms. Therefore, a detailed clinical examination of these animals is important.

Tumors and tumor-like lesions in the mammary gland of 24 pet rabbits: a histomorphological and immunohistochemical characterization.

The aim of this retrospective study (2004-2011) was to examine mammary tumors and tumor-like lesions in 24 pet rabbits by histopathology and immunohistochemistry. Rabbits were aged 2 to 8 years. Seventeen were female and 7 female-spayed. Diagnosed tumor-like lesions were lobular hyperplasia (2 rabbits) and multiple cysts (10 rabbits). Tumors included cystadenoma (7 tumors; 3 rabbits), intraductal papilloma (2 tumors; 1 rabbit), intraductal papillary carcinoma (1 tumor), adenocarcinoma (14 tumors; 13 rabbits), adenosquamous carcinoma (2 tumors; 2 rabbits), and matrix-producing carcinoma (1 tumor). The most frequently diagnosed lesion was invasive carcinoma (n = 17). Ten rabbits had several lesions. Immunohistochemistry for calponin and p63 showed that the diagnosed tumor-like lesions, benign tumors, and noninvasive carcinoma had a peripheral myoepithelial layer that was lacking in the invasive carcinomas. In 13 of 14 (93%) of the invasive carcinomas, however, there were variable numbers of calponin- and/or p63-immunopositive cells ranging from 0.1% to 40% with morphological features of either retained nonneoplastic myoepithelial cells or neoplastic epithelial cells with a myoepithelial differentiation. Tumor recurrence was reported in the rabbit with the matrix-producing carcinoma and in 3 rabbits with mammary adenocarcinomas displaying ≥20 mitotic figures in 10 high-power fields and high numbers of neoplastic cells with a myoepithelial differentiation (19%-39%). The rabbit with the matrix-producing mammary carcinoma developed cutaneous metastases confirmed by histopathology. This study shows that different types of mammary tumor-like lesions and tumors can occur in pet rabbits.

Comparison of cytological and histological examinations in different locations of the equine uterus-an in vitro study.

Endometritis constitutes a major problem in managing broodmares. The histological occurrence of polymorphonuclear neutrophils (PMNs) in the stratum compactum of the endometrium is accepted as the reference standard to diagnose endometritis in mares. The objective of this study was to determine the distribution of PMNs within different sampling locations of the uterus by cytological examinations and to compare it with PMN numbers in endometrial biopsies of the corresponding location. Cytological and endometrial samples were obtained from 37 uteri within 2 ± 1 hours after slaughter through small incisions from five different, predefined locations of each uterus. The cytological samples were smeared on microscopic slides, stained, and classified as negative (<2% PMNs) or positive (≥2% PMNs) for endometritis. Histologically, the numbers of PMNs were counted in three high power fields by an experienced pathologist and classified as positive for this type of endometritis if ≥5 PMNs occurred in three high power fields (×40 magnification). The biopsies were also evaluated for lymphoplasmacellular endometritis, periglandular fibrosis (endometrosis), and angiosclerosis. The prevalence of positive cytological and histological samples was 14.6% and 17.8%, respectively. A fair agreement between the two diagnostic methods could be detected (k = 0.29; P < 0.01). The numbers of PMNs differed between the sampling locations, resulting in positive and negative locations within a positive scored uterus, in both cytologically positive scored uteri (8/10) and histologically positive scored uteri (13/14). No significant differences were found in PMN numbers in the different locations, either the cytological (P = 0.78) or histological (P = 0.79) examination. Additionally, no significant differences were observed in the assessment of endometrosis (P = 0.96) and angiosclerosis (P = 0.67) within the locations. In conclusion, PMN numbers of a cytological examination of the endometrium showed fair agreement to the occurrence of PMN in the stratum compactum of the histological examination at the same sampling location. Although variations were found in the number of PMN using both methods (cytology and histology), statistically significant differences were not detected within the different locations (P = 0.78; P = 0.79), implyingies that the decision to take more than one sample should be critically considered. Additional research is warranted to determine the number of sampling locations necessary for reliable examination results.

Canine splenic haemangiosarcoma: influence of metastases, chemotherapy and growth pattern on post-splenectomy survival and expression of angiogenic factors.

Splenic haemangiosarcomas (HSAs) from 122 dogs were characterized and classified according to their patterns of growth, survival time post splenectomy, metastases and chemotherapy. The most common pattern of growth was a mixture of cavernous, capillary and solid tumour tissue. Survival time post splenectomy was independent of the growth pattern; however, it was influenced by chemotherapy and metastases. Immunohistochemical assessment of the expression of angiogenic factors (fetal liver kinase-1, angiopoietin-2, angiopoietin receptor-2 and vascular endothelial growth factor A) and conventional endothelial markers (CD31, factor VIII-related antigen) revealed variable expression, particularly in undifferentiated HSAs. Therefore, a combination of endothelial markers should be used to confirm the endothelial origin of splenic tumours.

GM2 gangliosidosis in an adult pet rabbit.

A 1.5-year-old neutered male rabbit was presented with chronic nasal discharge and ataxia. Rapid progression of neurological signs was noted subsequent to general anaesthesia and the rabbit was humanely destroyed due to the poor prognosis. At necropsy examination there were no gross changes affecting the brain or spinal cord. Microscopical examination revealed that the perikarya of numerous neurons in the brain and spinal cord were distended by the intracytoplasmic accumulation of pale, finely granular to vacuolar material. Transmission electron microscopy showed this to be composed of concentric membranous cytoplasmic bodies. Thin layer chromatography revealed elevation of GM2 ganglioside in the brain of this rabbit compared with that of an unaffected control rabbit. Enzymatically, there was markedly reduced activity of tissue ß-hexosaminidase A in brain and liver tissue from the rabbit. This was a result of an almost complete absence of the enzymatic activity of the α-subunit of that enzyme. These findings are consistent with sphingolipidosis comparable with human GM2 gangliosidosis variant B1.

Nonsuppurative encephalitis in a dog.

A 4-year-old male German Hunting Terrier presented with tremor, dyspnea, trismus, spasms of the musculature of the larynx and pharynx, and hypothermia and subsequently died despite intensive clinical care. Prior clinical signs included vomitus and diarrhea. Microscopic examination of the brain revealed a multifocal nonsuppurative brain stem encephalitis; a few intralesional neurons contained intranuclear inclusions. By immunohistochemistry, Aujeszky disease virus (Suid herpesvirus 1) antigen was detected in neurons in the brain and in ganglion cells of the trigeminal ganglia. Viral culture of brain tissue confirmed the presence of Aujeszky disease virus. Histopathologic findings in the brain with the identification of Aujeszky disease virus by immunohistochemistry and polymerase chain reaction are consistent with Aujeszky disease virus-induced encephalitis. Sequencing revealed a 100% homology of the isolated Aujeszky disease virus with Aujeszky disease virus isolates of wild boar from Eastern Germany.

[Clinical signs, diagnostic approach and therapy for the so-called ovarian remnant syndrome in the bitch]. / Klinische Symptomatik, diagnostische Vorgehensweise und Therapie beim so genannten Ovarrest-Syndrom der Hündin.

The ovarian remnant syndrome arises as a consequence of incomplete ovariectomy or ovariohysterectomy. Remnant ovarian tissue which has been left mostly unintentionally in the bitch may show endocrine activity a few weeks to several years after surgery, provoking a variety of clinical signs. The majority of affected bitches return to heat, in other cases signs of pseudopregnancy and endometritis may be observed. Occasionally, bitches with unclear clinical signs are presented with the suspicion of an inactive ovarian remnant. The following article intends to place the origin of the ovarian remnant syndrome into a factual context regarding the responsibility of the veterinarian and to demonstrate a reasonable diagnostic procedure according to the respective clinical signs. In this regard, the clinical-gynaecological examination, including vaginal cytology, must receive high priority, with the addition of progesterone analysis in peripheral blood plasma or serum if required. Using these combined diagnostic tools, ovarian remnants in stages of endocrine activity (follicular and luteal phases as well as cystic or tumourous ovarian tissue) can be easily unequivocally diagnosed. The application of a GnRH-stimulation test is only reasonable in bitches in which clinical signs are missing. In this context, the usefulness of semi-quantitative LH-assays is also discussed.