Endometrial biopsy in Holstein-Friesian dairy cows. I. Technique, histological criteria and results.

Endometrial biopsies were taken for histological assessment from 97 cows which calved in a commercial dairy herd between April and August 1984. Sixty-two cows were biopsied at both day 26 and 40 postpartum, 23 cows at only day 26, and 12 at day 40 only. Subjective and quantitative histological criteria were assessed. Ninety-five percent of biopsies were adequate for at least subjective assessment. The distribution of criteria within each horn-day category, as well as combined readings by day and by gravid or nongravid horn were computed and significant differences noted. There was more severe inflammation and more segmented cells at day 26 than 40 postpartum, and in the gravid compared to the nongravid horn. The distribution patterns for the criteria examined provide an overview of histological characteristics in this group of postpartum cows.     

Endometrial biopsy in Holstein-Friesian dairy cows. III. Bacteriological analysis and correlations with histological findings.

This study examines the results of bacterial culture from 159 endometrial biopsy samples from 97 commercial dairy cows and correlations between bacteriological and histological findings. Bacteria were isolated from approximately 80% of biopsies taken at day 26 and day 40 postpartum. Eleven percent of biopsies were positive for both aerobic and anaerobic culture. Streptococci, Escherichia coli and Actinomyces pyogenes were the most common isolates. Isolation of A. pyogenes from a biopsy at day 26 was positively correlated with isolation of anaerobic bacteria and segmented cell inflammation in the same biopsy, and with subsequent isolation of A. pyogenes at day 40. There was a strong association between isolation of A. pyogenes and anaerobes at day 26 with increased uterine lesions at day 40. Isolation of alpha hemolytic streptococci (AHS) was negatively correlated with isolation of A. pyogenes and with inflammation. Actinomyces pyogenes and AHS showed opposite associations with mononuclear cell inflammation and lymphocytic foci.     

Skin biopsies predict acute graft-versus-host disease after small bowel transplantation in pigs

Intestinal transplantation is being increasingly performed to treat patients with irreversible intestinal failure. The major cause of intestinal graft failure is graft-versus-host disease (GVHD) that represents a life-threatening complication after small bowel transplantation (Itx). The purpose of this study was to assess the diagnostic and prognostic value of skin biopsy histological changes for acute GVHD after Itx in pigs. Thirty-four Large White pigs were divided into three groups: Group 1 with Itx only, Group 2 with Itx and donor bone marrow infusion (Itx BM) and Group 3 (control group - before the operation). Animals received tacrolimus-based immunosuppression from day 0 to day 30 postoperatively. Skin and small bowel biopsies were histologically assessed, analysed and classified from grade 1 to 4 on postoperative days 15, 30, 45 and 60. There was a strong correlation between the histological grading values of skin biopsy changes and the histological grading values of small bowel biopsy changes (Kendall's tau_b is 0.855 for the Itx group and 0.730 for the Itx BM group). The significant correlation found between skin and small bowel histological changes suggests the prognostic value of skin biopsies after Itx. In conclusion, our findings emphasise the diagnostic and prognostic value of skin biopsy analysis for acute GVHD after Itx.     

Consequences of Intrauterine Enrofloxacin Infusion on Mare Endometrium

Endometritis is an important cause of infertility in mares. Enrofloxacin is a broad-spectrum antibiotic to which most equine endometritis pathogens are not resistant. The objective of this study was to determine whether enrofloxacin is safe to use as a conventional intrauterine infusion treatment. Nine healthy mares received intrauterine infusions of enrofloxacin (Baytril 100, 100 mg/mL, Bayer Health Care LLC, Animal Health Division) at 2.5 mg/kg daily for 3 days. Ultrasonographic examination and vaginal examinations were performed during the study. Endometrial biopsies were performed before treatment (S0) and 24 hours after the last treatment (S1) to evaluate acute effects. For evaluating chronic effects, biopsies were performed at 14 days (S2) and 60 days posttreatment (S3). Biopsies were graded histologically by the Kenny and Doig category scale. Difference in histological biopsy grade before and after treatment was compared between biopsies by using a repeated-measures one-way analysis of variance. and significant changes in grades were used to assess treatment effects. The vaginal and ultrasonographic examination after intrauterine infusion of enrofloxacin showed that all mares had severe purulent vaginitis and uterine fluid accumulation of ≥2 cm, with ≥1.5-cm thickening of the endometrial wall which persisted in most mares until the end of the study. Histologically, there was acute endometrial ulceration, necrosis, and hemorrhage in biopsy S1 in all mares, categorized as grade III. In biopsy S2, most mares developed fibrosis and inflammation graded as IIb (four of nine mares) or III (four of nine mares). In biopsy S3, fibrosis was extensive and had variable inflammation, graded as IIb (two of nine mares) or III (five of nine mares), with some mares healing to grade IIa (two of nine mares). There was an overall worsening of endometrial biopsy grade from I to III at S3 compared with S0 (P < .001). These results confirm that enrofloxacin is not suitable for conventional intrauterine infusion treatment in mares.     

A quantitative study of the histological morphology of the endometrium of normal and barren mares.

The density of uterine glands, height of surface epithelium, numbers of hemosiderin laden macrophages, inflammatory cells and layers of periglandular fibrosis were evaluated in uterine biopsies from 40 mares. These features were found to be highly variable in normal equine endometrium. Minor pathological changes appeared to be masked by this normal variability. Atrophy of uterine glands was recognized in mares which had been barren for more than three years. No significant differences were found between barren and normal mares in the height of epithelium, number of hemosiderin laden macrophages, inflammatory cells or layers of collagen surrounding glands in the superficial portion of endometrium. The number of layers of fibrosis surrounding glands in the deep part of lamina propria were found to be correlated with years of barrenness. This finding appears to have prognostic potential.     

Influence of inflammation and coat type on oestrogen receptor immunohistochemistry

The purpose of this study was to evaluate oestrogen receptor alpha staining in a variety of breeds and skin conditions. The influence of inflammation and coat type on the presence and intensity of oestrogen receptor alpha staining was evaluated. Approximately 1700 haematoxylin and eosin (H&E)-stained slides of skin biopsies were screened for presence of small hair bulbs. Slides from 94 cases were submitted for oestrogen receptor alpha immunohistochemistry. H&E-stained skin biopsy tissues were examined for inflammation and hair follicle stages. Oestrogen receptor alpha staining characteristics of telogen follicles, flame follicles, large anagen bulbs, small hair bulbs and early anagen hairs (capped bulbs) were recorded. To assess the influence of inflammation and coat type on oestrogen receptor staining of hair follicle types, chi-square tests, Fisher's exact tests and logistic regression models were performed. Slides were classified as inflammatory (65) and noninflammatory (29). There were no statistically significant differences in oestrogen receptor staining when comparing inflammatory to noninflammatory skin biopsies or skin biopsies from dogs with different coat types. A subset of 13 noninflammatory biopsies from alopecic skin was identified. There was a statistically significant increase in the number of flame follicles in this subset as compared to all others. Comparison of oestrogen receptor staining of hair follicle types from these biopsies and all other biopsies revealed a statistically significant increase in the number of mature telogen hair follicles stained in this subset. No statistical difference in staining of early follicle stages was noted. Therefore, the oestrogen receptor is unlikely to be the controlling factor for the transition from telogen to anagen in the dog.     

Evaluation of the tissue reactions in the skin and body wall of koi (Cyprinus carpio) to five suture materials

Five different suture materials (silk, monofilament nylon, polyglyconate, polyglactin 910, and chromic gut) were placed in the skin and body wall of 10 Doitsu (scaleless) koi (Cyprinus carpio). After seven days the sutures were retrieved from five of the fish in 5 mm and 6 mm punch biopsies, and after 14 days they were retrieved in the same way from the other five. The tissue reactions were evaluated by gross visual inspection and by histological examination. The total inflammatory reaction was graded on a scale from 0 (no inflammation) to 5 (severe inflammation). The synthetic suture materials generally induced a moderate inflammatory reaction that decreased after seven days. After 14 days the superficial reaction to monofilament nylon was substantial, and the tissue reactions to the organic suture materials were slightly greater than the reactions to the synthetics. The inflammatory response to silk was greater after 14 days than after seven, and chromic gut induced a moderately severe inflammatory response after seven days; the chromic gut sutures fell out before the biopsies were taken after 14 days. The organic materials induced intense inflammatory reactions which did not subside if the suture remained in the tissue.     

Serial percutaneous testicular biopsy in the Beagle dog

The effects of serial percutaneous testicular biopsy on testicular size, semen characteristics and plasma testosterone have been assessed. Testicular atrophy occurred in one dog after the first biopsy, but apparently there were no permanent deleterious effects on semen characteristics or testosterone concentrations. The histological changes 72 hours and 14 days after bilateral testicular biopsy were examined in one dog. Initial haemorrhage and necrosis with subsequent fibrosis and atrophy occurred at the biopsy site. The surrounding tissue remained normal in appearance. The technique may be worthwhile for establishing diagnostic criteria in cases of male canine infertility. Not enough tubules are obtained in circular cross-section to allow detailed histomorphometric analysis of spermatogenesis under laboratory conditions.     

Histological Changes in the Uterus during Postpartum in the Mare

An histological study of the postpartum period in 29 mares was carried out. Uterine biopsies were taken daily during the first 10 days postpartum in a total of 87 samples. At day 0, equine endometrium was characterized in the surface by the presence of regularly ordered microcaruncles; the stratum spongiosum was oedematous and contained distended and scarce glands. Degenerative changes in microcaruncles and endometrial glands were present on day 1 postpartum. The epithelium of the microcaruncles from 2 to 5 days postpartum showed cytoplasmic vacuolization, karyorrhexis and an inflammatory reaction with neutrophils and phagocitic cells. On day 7 postpartum, the histology of the endometrium was similar to the normal proestrus with cuboidal luminal epithelium and an oedematous stromal tissue. The changes are usually completed within days 9 and 10 postpartum with the histologically typical appearance of estrus in mares.     

Morphologic evaluation of acute endometritis in mares with differing resistance to uterine infections

The study was designed to determine differences between normal mares and mares with endometrial pathology in the inflammatory response after bacterial challenge. Six normal mares (biopsy category I) and 4 mares with pathological endometrial changes (biopsy category II) were given an intrauterine infusion of β-hemolytic streptococci on the second day of estrus. All mares had a similar kind of inflammatory response after the bacterial inoculation as assessed by rectal and vaginal examinations. There were no significant differences in the amount of discharge, uterine tone, uterine size and cervical relaxation between the groups. Leukocytic response, as determined by endometrial smears and biopsies, was of the same magnitude in both groups. Two mares from the pathological group were not able to eliminate the infection, but had vaginal discharge and bacteriologically positive uterine swabs until the end of the experiment. It is concluded that the inability of some mares to clear uterine infections cannot be explained by a deficient inflammatory response.     

Comparison of Histopathologic Findings in Duodenal and Ileal Endoscopic Biopsies in Dogs with Chronic Small Intestinal Enteropathies

Background

The current tendency when investigating dogs with chronic upper gastrointestinal signs is to perform endoscopy and biopsy only the duodenum. This approach could lead to overlooking important ileal lesions and affect the clinical management.

Objectives

To compare concurrent duodenal and ileal endoscopic biopsies in dogs with chronic enteropathies and evaluate their correlation with clinicopathologic findings.

Animals

Thirty‐eight dogs with chronic enteropathies.

Methods

Duodenal and ileal biopsies were retrospectively reviewed. Nine histologic variables, 5 structural (villous stunting, epithelial injury, crypt distension, lacteal dilatation, and mucosal fibrosis) and 4 inflammatory (intraepithelial lymphocytes, lamina propria lymphocytes and plasma cells, eosinophils, and neutrophils) were scored. Clinical severity scores and relevant clinicopathologic variables were evaluated.

Results

There was only slight agreement between duodenal and ileal histologic scores (κ = 0.003). There was slight agreement between the presence of any of the morphological and inflammatory variables, with the exception of mucosal fibrosis (κ = 0.44). Statistically significant correlation was found between clinical severity and duodenal crypt distension (P = .031), ileal lacteal dilatation (P = .038), and ileal mucosal lymphoplasmacytic inflammation (P = .035). A significant correlation was found between hypoalbuminemia and ileal lacteal dilatation (P = .033) and number of ileal intraepithelial lymphocytes (P = .019). A statistically significant correlation was found between hypocobalaminemia and number of ileal intraepithelial lymphocytes (P = .012).

Conclusions and Clinical Importance

When investigating dogs with chronic upper gastrointestinal signs, the collection of concurrent duodenal and ileal endoscopic biopsies is recommended.     

Preliminary study of tendon biopsy in the horse

A series of experimental tendon biopsies is described. Three biopsies were taken from the lateral digital extensor tendon (LDET) and three from the superficial flexor tendon (SFT). The LDET biopsies resulted in little discomfort whereas the SFT biopsies led to temporary lameness. The tendons were examined histologically up to 99 days after the biopsies were removed. In all of the tendons the defect filled with granulation tissue which subsequently became organised as a longitudinally orientated collagenous scar tissue. In this small series of biopsies the histological effects of the biopsy persisted longer in the SFT than in the LDET. It is concluded that tendon biopsy cannot be advocated for safe routine use in the horse.     

Inflammatory myopathies.

Inflammatory myopathies are the result of infiltration of inflammatory cells into striated muscle, with or without an association with an underlying cause. Two broad classifications are IIMs and secondary inflammatory myopathies associated with other diseases. Standard diagnostic criteria for inflammatory myopathy include the presence of weakness or loss of specific muscle group function, an increase in CK, EMG changes associated with muscle membrane instability, and histologic evidence of inflammation. Not all these criteria, however, must be present. Fresh-frozen biopsy from two proximal muscles is recommended for biopsy confirmation. IIM can either focally affect head or neck muscles or be more diffuse. MMM is an immune-mediated disease characterized by a humoral antibody produced against the unique type IIM and type I variant mvofibers of masticatory muscles of dogs, which causes inflammation and loss of function of the muscles of mastication. Idiopathic polymyositis can affect focal muscle groups (extraocular, laryngeal) or present as multifocal or diffuse involvement of skeletal muscle in the cat and dog. Familial canine DM is an inflammatory disease of the striated muscle, skin, and vasculature in young Collies, Shetland Sheepdogs (Shelties), and, rarely, Collie-crossbred dogs. Immunosuppressive therapy is the key to successful treatment. Protozoal parasitic myopathies are the most common cause of clinically relevant secondary inflammatory myopathies. The degree of systemic involvement is often the limiting factor to successful treatment. Early recognition of the clinical signs for proper diagnostic testing and institution of appropriate therapy can result in a rewarding outcome in treating inflammatory myopathies in the cat and dog.     

Ultrasound- guided catheter biopsy of the lower urinary tract: technique and results in 12 dogs

Ultrasound-guided catheter biopsy of lesions affecting the lower urinary tract was attempted in 12 dogs with mucosal lesions affecting the bladder [nine] or urethra (three). Histological biopsies were obtained by catheter biopsy in 10 dogs, enabling diagnosis of transitional cell carcinoma in five, papilloma in two, prostatic carcinoma in two and chronic cystitis in one. Cytological samples alone were obtained in two dogs, one of which enabled a diagnosis of transitional cell carcinoma; the other contained evidence of haemorrhage and inflammation, but squamous cell carcinoma was found in a subsequent exci-sional biopsy. Intravesicular haemorrhage after biopsy was observed ultrasonographically in two dogs. Ultrasound guidance enables accurate determination of biopsy catheter position. The size of biopsies obtained by this method may limit the accuracy of histological diagnosis.     

Follow-up studies by peroral gastric biopsies and necropsy in vomiting dogs.

The results of follow-up studies in 139 vomiting dogs are presented. Follow-up studies were performed by biopsies in 34 dogs, by biopsies and necropsy in six dogs and by necropsy only in 99 dogs. The times between the first and the last series of biopsies varied from three to 1042 days and from one to 656 days between the first series of biopsies and necropsy. From the 55 dogs with gastritis in the first series of biopsies, 35 also showed gastritis in the following biopsies or at necropsy. These were mainly severe types of gastritis such as diffuse, hypertrophic or atrophic. Ten dogs with superficial gastritis showed no gastric changes at necropsy, two dogs had edema only and three dogs had gastric changes other than gastritis, such as multiple polyps. In general, carcinoma and lymphosarcoma were found in the biopsies as well as at necropsy, but in three cases of terminal carcinoma only gastritis had been diagnosed initially. In 35 dogs the first series of gastric biopsies showed no pathological changes, but in 22 of these dogs gastritis, ulceration, fibrosis, atrophy, gastric dilation with local necrosis, and perforation or lymphosarcoma of the submucosa were found in the second series of biopsies or at necropsy. Several dogs which did not have gastric changes at necropsy had enteritis or intestinal lymphosarcoma.     

Comparison of biopsy samples obtained using standard endoscopic instruments and the harmonic scalpel during laparoscopic and laparoscopic-assisted surgery in normal dogs

Objectives— To compare tissue specimens of canine abdominal organs collected by standard techniques (ST) and harmonic scalpel (HS) and to determine the effect of each technique on wound healing.
Study Design— Experimental.
Animals— Twelve dogs.
Methods— Paired biopsy samples (ST, HS) were collected from liver, spleen, adrenal gland, pancreas, stomach, jejunum, and bladder using laparoscopic or laparoscopic-assisted methods. Hemorrhage at biopsy sites was assessed (present or absent). Specimens were evaluated for diagnostic quality and histologic changes. Dogs were necropsied at 4 or 14 days postoperatively.
Results— HS incision caused less frequent hemorrhage of all organs except spleen. HS specimens had more coagulation necrosis than ST specimens, but both techniques yielded specimens that were sized similarly and were of diagnostic quality. At necropsy, HS biopsy sites of the stomach, jejunum, bladder, adrenal gland, and pancreas were more hyperemic than ST sites. There were more adhesions at jejunal and pancreatic ST biopsy sites. HS biopsy sites had more coagulation necrosis at days 4 and 14 postoperatively. Differences in fibrin deposition, inflammation, and fibrosis were present at biopsy sites of some organs at days 4 or 14 and in comparisons between days 4 and 14.
Conclusions— Diagnostic quality biopsy specimens were obtained with HS and ST. Although HS-induced gross and histologic changes during the first 2 postoperative weeks, no clinical complications were observed.
Clinical Relevance— Both HS and ST can yield specimens with minimal hemorrhage and HS resulted in no apparent postoperative problems in normal dogs. Although HS caused more inflammation and adhesions at biopsy sites of the pancreas, adrenal gland, and jejunum, no clinical complications occurred.     

The influence of multiple liver biopsies on hematologic and serum biochemical values of sheep

A rapid, simple liver biopsy technique for sheep was developed to obtain in situ liver samples for evaluation of histological tissue, enzyme assay, and ultrastructural changes that might occur during toxicological testing. The study was designed because repeated biopsies in the same individual could induce bias in hematologic, histological or serum biochemical parameters caused by the trauma of the biopsy procedure. In Experiment I, sheep were divided into a biopsied group (B), sham-operated group (SO), and a nonsurgical, untreated control group (C). Three liver biopsy samples per week were removed from each subject in Group B for five weeks. Biweekly blood samples were drawn for evaluation of hematologic and serum biochemical parameters. In Experiment II, Groups B and SO were dosed with CCl4 in order to validate the applicability of the biopsy procedure. Weekly biopsies were continued in Group B. Biweekly blood samples were evaluated for any hematologic or serum biochemical variations. Significant elevations of enzyme activities confirmed CCl4 hepatotoxicity in treated groups (B and SO) versus C group. Histological lesions in biopsy samples correlated closely with serum biochemical data in treated groups. It was concluded that the serial liver biopsy procedure causes negligible alterations in the hematologic, histological or serum biochemical parameters measured; is simple and rapid to perform, and provides a method for possible early detection of liver disease.     

Enterohepatic Helicobacter spp. in colonic biopsies of dogs: molecular, histopathological and immunohistochemical investigations

Enterohepatic Helicobacter spp. have been described colonizing the large intestine and liver of healthy and symptomatic subjects and are thought to have a role in the development of inflammatory bowel disease (IBD). The prevalence of enterohepatic Helicobacter spp. infection in dogs is largely unknown and to our knowledge there are no data about their potential pathogenic role. In light of these considerations, the aims of this study were (i) to assess the prevalence of enterohepatic Helicobacter spp. in colonic biopsies of symptomatic pet dogs and (ii) to evaluate a possible association between Helicobacter spp. colonization status (heavily colonized, poorly colonized and uncolonized biopsies) and histological lesions. Colonic biopsies from 27 pet dogs of different ages were evaluated by family Helicobacteraceae and enterohepatic Helicobacter spp. PCR, histology, and immunohistochemistry for the in situ detection of Helicobacter spp. organisms. 85% and 52% of colonic biopsies were positive by Helicobacteraceae and enterohepatic Helicobacter spp. PCR, respectively. Immunohistochemistry revealed Helicobacter spp. were localized both in the superficial mucus (55%) and within intestinal crypts (33%). Dogs with heavy enterohepatic Helicobacter spp. colonization were significantly younger and had a higher level of mucosal fibrosis/atrophy than dogs with uncolonized or poorly colonized biopsies (p<0.05). These findings contribute to widen current knowledge regarding canine enterohepatic Helicobacter spp., suggesting the infection is rather common in dogs and acquired at an early age. Furthermore, heavy colonization of colonic crypts is associated with chronic inflammatory lesions (fibrosis/atrophy), supporting the role of enterohepatic Helicobacter spp. in the development of canine IBD.     

Uterine bacteriology, histology, resumption of ovarian activity and granulocyte function of the postpartum cow in different milking frequencies

The postpartum uterine bacteriology, histology, resumption of ovarian activity and polymorphonuclear granulocyte (PMN) number and function in 18 Swedish dairy cows were studied. Cows were milked either 2x (n = 9) or 3x per day (n = 9). Endometrial biopsy samples for bacteriological and histological investigations were collected during 8 weeks postpartum, starting within one week after calving. Milk samples for progesterone determination were collected twice a week until the cows had shown normal reproductive cyclicity. Blood samples for granulocyte function (phagocytic capacity and total number) were collected from each animal on the same days as when the biopsies were obtained. All animals in both groups were free from bacteria at the latest after 6 weeks post-partum and there was no difference regarding bacterial elimination and bacterial species between milking groups. No difference regarding uterine histology between milking groups was seen. In both groups, 8 cows had normal to slight infiltration of leukocytes in the endometrium at the end of sample collection. No changes in granulocyte function could be seen in the 2 milking groups. Resumption of ovarian activity was detected on day 45.6 +/- 9.3 (mean +/- SD) postpartum in the 2x milking group and 36.6 +/- 9.0 (mean +/- SD) post-partum in the 3x milking group (p = 0.05). Based on our findings, an increased milking frequency from 2 to 3 times a day did not influence the uterine function postpartum.     

Ultrasonographic and quantitative histologic assessment of sequelae to testicular biopsy in stallions.

A sample of testicular parenchymal tissue, approximately 2 x 7 x 7 mm, was aseptically removed from 1 testis in each of 9 stallions on day 0. Slight to moderate hemorrhage from the tunica albuginea was observed in 8 stallions, but bleeding from the parenchyma was detected in only 2 stallions. Stallions were castrated 27 days later. Normal development of granulation tissue was evident at the biopsy site, but hematomas were not observed. In situ measurement of the widths of the right and left testes, total scrotal width, and evaluation of testicular echogenicity during ultrasonography were variables used to monitor changes in the testicular parenchyma from 14 days before biopsy through 27 days after biopsy. The control testis was consistently larger than the biopsied testis, except for day 3. Ultrasonography revealed signs of a localized change in the parenchyma of the biopsied testis in 4 stallions, but each lesion decreased in size by day 27. Tissues removed during biopsy enabled an excellent appraisal of spermatogenesis at that time. Detailed examinations of seminiferous tubules in the testes were performed to assess for damage to testicular function. At castration, samples were taken from 6 sites in each testis. Quantitative histologic evaluations of testicular tissues revealed low numbers of spherical spermatids and pachytene spermatocytes in biopsied testes, compared with control testes. It was concluded that there was a transitory increase in degeneration of preleptotene spermatocytes and B spermatogonia at the time of biopsy. A mild inflammatory response at the biopsy site in some testes was evidenced by an increased number of leukocytes at the biopsy site and at a dorsal site.(ABSTRACT TRUNCATED AT 250 WORDS)