Results of analyses and bacterial cultures of urine specimens obtained from clinically normal cats by three methods

Six urine specimens were obtained from each of 6 male and 6 female cats in a 3-week period. The first and last specimens from each cat were obtained by cystocentesis, 2 were obtained by urethral catheterization, and 2 were caught during voiding stimulated by manual bladder compression. Quantitative urine cultures did not reveal bacteriuria in specimens obtained by cystocentesis, and urinary tract infection did not develop during the study. Bacteria were found in 3 (25%) of 12 urine specimens obtained by catheterization of males and in 1 (8%) of 12 specimens obtained by catheterization of females. Magnitude of bacteriuria in specimens obtained by catheterization was 10 to 1,000 organisms/ml. Bacteria also were found in specimens obtained during voiding. Each of 11 (100%) cleanly caught specimens obtained from males and 7 (58%) of 12 voided specimens from females contained bacteria. Magnitude of bacteriuria in voided specimens was usually 100 to 10,000 organisms/ml, but 3 voided specimens contained greater than 10,000 organisms/ml. Many bacteriuric specimens contained more than 1 type of organism; however, bacteria that might be suspected of causing urinary tract infections in cats were found frequently. Urinalyses were performed on 66 specimens. Completeness of urinalyses depended on the volume of specimens available, but results were normal except for evidence of hematuria in a few specimens obtained by cystocentesis or catheterization. Hematuria was usually mild and was attributed to hemorrhage caused by minor urinary tract trauma during urine collection.     

Simultaneous detection and strain differentiation of Mycobacterium bovis directly from bovine tissue specimens by spoligotyping

Culture of Mycobacterium bovis is used routinely to support field diagnosis of bovine tuberculosis; however, this method is slow. Rapid detection and strain-typing of M. bovis directly from 37 lesioned bovine lymph node specimens was performed by the polymerase chain reaction (PCR) based method, spoligotyping. Mycobacterial DNA was extracted from the specimens using a nucleic acid sequence capture technique. Two sets of specimens were tested, the first set comprising 16 decontaminated tissue homogenates from lesioned lymph node specimens which had been processed for BACTEC culture and a second set of 21 non-decontaminated lesioned lymph node specimens. Both sets of specimens had been frozen before analysis. Sequence capture PCR enabled detection and strain-typing of M. bovis directly from 15 of the 16 decontaminated homogenates and all 21 of the non-decontaminated tissues. Four spoligotype (ST) patterns were obtained from each set; ST1, ST2, ST3 and ST16 were detected in the decontaminated specimens and ST1, ST2, ST11 and ST14 in the non-decontaminated specimens. For both sets of specimens, ST1 was the predominant strain type detected. ST patterns obtained from the BACTEC cultures of the decontaminated specimens were in agreement with those obtained directly from the tissue. The sensitivity of detection by sequence capture-PCR compared very favourably with that of BACTEC culture. ST patterns were obtained directly from tissues of 34 of the 35 culture positive specimens and the two culture negative specimens. DNA extraction from the 21 non-decontaminated specimens involved an initial stomaching treatment. An assessment of sequence capture on both liquid alone and liquid and tissue homogenate combined, following stomaching, indicated that PCR was less successful on the liquid component alone.     

Diagnostic comparison of needle and wedge biopsy specimens of the liver in dogs and cats

OBJECTIVE: To compare morphologic diagnoses determined from needle biopsy specimens obtained from the livers of dogs and cats with morphologic diagnoses determined from wedge biopsy specimens. DESIGN: Prospective study. ANIMALS: 124 dogs and cats. PROCEDURE: 2 needle biopsy specimens were obtained from each animal; wedge biopsy specimens were obtained from the same liver lobe during laparotomy or postmortem examination. Histologic features were scored independently by 3 individuals; a morphologic diagnosis was rendered after histologic features were scored. Cases were included only if at least 2 of the 3 examiners agreed on the morphologic diagnosis; the definitive diagnosis was considered to be the morphologic diagnosis rendered for the wedge biopsy specimen. Physical characteristics (length, width, surface area, degree of fragmentation, and number of portal triads for needle biopsy specimens and surface area for wedge biopsy specimens) were determined. RESULTS: Definitive diagnoses included hepatic necrosis (n = 10), cholangitis-cholangiohepatitis (13), chronic hepatitis-cirrhosis (12), canine vacuolar hepatopathy (11), portosystemic vascular anomaly-microvascular dysplasia (17), neoplasia (10), miscellaneous hepatic disorders (18), and no hepatic disease (33). For individual examiners, the morphologic diagnosis assigned to needle biopsy specimens agreed with the morphologic diagnosis assigned to wedge biopsy specimens for 56 and 67% of the specimens. All 3 examiners agreed on the morphologic diagnosis assigned to needle and wedge biopsy specimens for 44 and 65% of the specimens, respectively. Morphologic diagnoses assigned to needle biopsy specimens concurred with the definitive diagnosis for 59 of 124 (48%) animals. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that needle biopsy specimens of the liver from dogs and cats must be interpreted with caution.     

Polymerase chain reaction-based restriction fragment length polymorphism pattern of porcine reproductive and respiratory syndrome virus directly from lung tissues without virus isolation in Korea.

Polymerase chain reaction (PCR)-based restriction fragment length polymorphism (RFLP) analysis was developed for directly typing porcine reproductive and respiratory syndrome virus (PRRSV) from lung specimens without virus isolation. Twenty nine lung specimens collected from postweaning pigs were isolated for PRRSV. When the PCR products from the 29 lung specimens were digested by the restriction enzymes MluI, HincII, SacII and HaeIII, the RFLP patterns from the 29 lung specimens matched with those from the corresponding PRRSV isolates from each pig. The results suggest that the PCR-based RFLP analysis method may be useful to distinguish PRRSV isolates directly from lung specimens without virus isolation.     

Mercury residues in goshawk (Accipiter gentilis) in 1966 and 1974 (author's transl)]

A comparative study of liver, muscle and kidney specimens from goshawks (Accipiter gentilis) shot in 1966 and 1974 respectively shows that a significant decrease in the mercury levels has taken place. The mean values in the liver specimens have changed from 2.27 mg/kg to 0.50 mg/kg, in the muscle specimens from 0.99 mg/kg to 0.20 mg/kg and in the kidney specimens from 3.06 mg/kg to 0.57 mg/kg. The main cause of this decrease is supposed to depend upon the recommendations since 1969 to avoid alkylmercurials as seed dressing.     

Histologic comparison of skin biopsy specimens collected by use of carbon dioxide or 810-nm diode lasers from dogs

OBJECTIVE: To compare histologic artifacts caused by carbon dioxide (CO2) or 810-nm diode surgical lasers used to obtain small biopsy specimens of skin from healthy dogs. DESIGN: Prospective study. ANIMALS: 4 dogs. PROCEDURE: 21 skin biopsy specimens were collected from each dog. Three biopsy specimens were obtained with a CO2 or an 810-nm diode laser at 3 operating settings each, and 3 biopsy specimens were obtained with a 6-mm biopsy punch instrument (controls). After processing, biopsy specimens were examined for artifacts related to laser-tissue interactions. Microscopically visible char was measured from the lateral edge of each specimen obtained with a laser. RESULTS: There were no significant differences among mean char distances in biopsy specimens obtained with the CO2 laser at various settings. Mean char distance was significantly greater in all skin biopsy specimens obtained with the diode laser, compared with those obtained with the CO2 laser. Mean char distance was significantly greater in biopsy specimens obtained with the 810-nm diode laser at high power, compared with biopsy specimens obtained with the 810-nm diode laser at low power. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that the CO2 laser caused less thermal injury at margins of skin biopsy specimens; therefore, if a surgical laser is used for removal of cutaneous masses or to obtain skin biopsy specimens, use of the CO2 laser is recommended. Veterinarians performing a biopsy by using a surgical laser should be aware that laser-induced artifacts may render small biopsy specimens useless for providing accurate histologic diagnosis.     

Polymerase chain reaction analysis for viruses in paraffin-embedded myocardium from dogs with dilated cardiomyopathy or myocarditis

OBJECTIVE: To perform polymerase chain reaction (PCR) analysis on paraffin-embedded myocardium from dogs with dilated cardiomyopathy (DCM) and dogs with myocarditis to screen for canine parvovirus, adenovirus types 1 and 2, and herpesvirus. SAMPLE POPULATION: Myocardial specimens from 18 dogs with an antemortem diagnosis of DCM and 9 dogs with a histopathologic diagnosis of myocarditis were evaluated. PROCEDURE: Paraffin-embedded myocardial specimens were screened for viral genome by PCR analysis. Positive-control specimens were developed from cell cultures as well as paraffin-embedded tissue specimens from dogs with clinical and histopathologic diagnoses of viral infection with canine parvovirus, adenovirus types 1 and 2, and herpesvirus. The histologic characteristics of all myocardial specimens were classified regarding extent, location, and type of inflammation and fibrosis. RESULTS: Canine adenovirus type 1 was amplified from 1 specimen from a dog with DCM. Canine parvovirus, adenovirus type 2, and herpesvirus were not amplified from any myocardial specimens. Histologic analysis of specimens from dogs with DCM revealed variable amounts of fibrosis; myocardial inflammation was observed in 1 affected dog. Histopathologic analysis of specimens from dogs with myocarditis disclosed variable degrees of inflammation and fibrosis. CONCLUSIONS AND CLINICAL RELEVANCE: Viral agents canine parvovirus, adenovirus types 1 and 2, and herpesvirus are not commonly associated with DCM or active myocarditis in dogs. Additional studies evaluating for nucleic acid from viruses that less commonly affect dogs or different types of infectious agents may be warranted to gain insight into the cause of DCM and myocarditis in dogs.     

Demonstration of bovine herpesvirus type 1 and Mannheimia haemolytica antigens in specimens stored for up to 22 months in buffered formalin

Bovine herpesvirus type 1 (BHV-1) and Mannheimia haemolytica antigens were demonstrated in lung tissues that were stored in 10% neutral phosphate buffered formalin for 1 to 22 months using the immunoperoxidase method. There were no differences observed in terms of labelling intensity and distribution of M. haemolytica antigens between specimens stored for 1 and 22 months. The labeling intensity in sections from 2-cm thick specimens was comparable to those from 0.2-cm thick specimens. There was no difference observed between pronase-treated and -untreated sections. However, for BHV-1, the labeling intensity in untreated sections was reduced in tissues that had been stored from 12 to 22 months. Sections from thin specimens stored in neutral buffered formalin for 22 months exhibited a stronger staining intensity than those from thick specimens.     

Clinical significance of aerobic bacterial flora of the uterus, vagina, vestibule, and clitoral fossa of clinically normal mares

Swab specimens for bacterial culture were obtained from the uterus, vagina, vestibule, and clitoral fossa of 48 mares that had normal reproductive tracts, no history of reproductive problems, and no inflammation on evaluation of endometrial biopsy. The mares were predominantly Thoroughbred and Standardbred. Swab specimens of the vagina were obtained through a sterile speculum; swab specimens of the uterus were obtained by use of a double-guarded, occluded culture instrument. Fifteen (31%) of the uterine swab specimens and 20 (42%) of the vaginal swab specimens yielded growth on aerobic culture; however, only 2 (4%) of the uterine swab specimens and 4 (8%) of the vaginal swab specimens yielded growth of more than 10 colonies. In contrast, 21 (44%) of the vestibular swab specimens and 45 (94%) of the clitoral fossa swab specimens had moderate (greater than 10 colonies in 1 quadrant) to heavy (colonies in 2 or 3 quadrants) growth of organisms on culture. Of organisms considered to be potential pathogens, Streptococcus zooepidemicus and Escherichia coli were found on bacteriologic culture of several clitoral fossa swab specimens and of some vestibular swab specimens. We did not isolate any potential pathogens from uterine or vaginal swab specimens. It appears that 1 to 10 colonies of nonpathogenic organisms could be recovered from the uterus in a substantial number of clinically normal mares even when double-guarded swabbing techniques are used, and we suggest that prebreeding culture requirements be modified to reflect this. Also, our findings indicate that the vulvovaginal fold, rather than the cervix, might be the major barrier to ascending bacterial contamination of the reproductive tract.(ABSTRACT TRUNCATED AT 250 WORDS)     

Detection of feline herpesvirus 1 DNA in skin biopsy specimens from cats with or without dermatitis

OBJECTIVE: To compare detection rates of feline herpesvirus 1 (FHV-1) DNA in skin biopsy specimens from cats with herpetic dermatitis, cats with nonherpetic dermatitis, and cats without dermatitis. DESIGN: Prevalence survey. Animals-5 cats (9 biopsy specimens) with herpetic ulcerative dermatitis, 14 cats (17 biopsy specimens) with nonherpetic ulcerative dermatitis, and 8 cats (21 biopsy specimens) without clinically apparent skin lesions. PROCEDURES: A single-phase PCR assay was used to detect FHV-1 DNA in biopsy specimens. Assay results were compared with results of histologic examination. RESULTS: FHV-1 DNA was detected in all 9 biopsy specimens from the 5 cats with herpetic dermatitis and in 1 of 17 biopsy specimens from the 14 cats with nonherpetic dermatitis, but was not detected in any of the 21 biopsy specimens from the 8 cats without dermatitis. When results of histologic examination were used as the gold standard, sensitivity and specificity of the PCR assay were 100% and 95%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Results confirmed that FHV-1 DNA can be detected in the skin of cats with herpetic dermatitis and suggest that the virus may play a causative role in the disease. In addition, the PCR assay may be useful in confirming a diagnosis of herpetic dermatitis.     

Corticocancellous Bone Biopsy from the 12th Rib of Standing Horses

Unicortical corticocancellous bone biopsy specimens 4.5 mm and 6.5 mm in diameter were obtained without long-term complications from the 12th rib in eight standing horses. However, the bone specimens were unsuitable for histologic or histomorphometric evaluation. In in vitro comparisons of biopsy specimens 6.5 mm and 12 mm in diameter, and of unicortical and transcortical biopsy specimens, 12 mm transcortical specimens yielded the most cancellous bone. Transcortical bone biopsy specimens 12 mm in diameter were obtained from eight horses by using power-assisted trephination. The surgical procedure was well tolerated, but some degree of pneumothorax in all horses was treated by aspiration of air from the thorax. Long-term complications were not observed.     

Comparison of two sample collection methods for quantitative bacteriologic culture of canine prostatic fluid

Ejaculate, urine, urethral swab specimens, and ultrasonography-guided small-needle prostatic cyst aspiration and/or tissue core biopsy specimens were collected for bacteriologic culture from 25 dogs in which prostatic disease was suspected on the basis of history, clinical signs of disease, or results of physical examination. The prostate gland in each dog was examined ultrasonographically, and the tissue core biopsy specimens were examined histologically and bacteriologically. Two methods were used to assess bacterial prostatitis. In 5 dogs (20%), bacteriologic culture results of paired urethral swab and ejaculate specimens differed from culture results of specimens obtained by needle aspiration of prostatic cyst fluid or tissue core biopsy. The prostate gland in 17 dogs had 1 or more cystic, fluid-filled structures (0.5 to 4.0 cm in diameter). Ultrasonographic appearance of the prostate gland did not have obvious correlation with culture results from dogs of the study. Histologic results of prostatic tissue core biopsy specimens correlated well with culture results.     

A modified biopsy technique to improve histopathological evaluation of avian skin

Skin biopsies are a viable diagnostic tool in avian dermatology, however, the thinness of avian skin makes it difficult to prevent rolling and contraction of skin biopsy specimens during collection and fixation. The difficulty orienting such rolled samples during processing ultimately interferes with the establishment of a histopathological diagnosis. We describe a modified skin biopsy procedure for obtaining avian skin biopsy specimens. In this technique nontranslucent self-adhesive tape (Scotch tape) was attached to skin biopsy sites before obtaining skin biopsies using a standard skin biopsy punch instrument. A total of 23 skin biopsy specimens were obtained: 15 from nonfeathered skin of 12 normal Hispaniolan parrots, 3 from feathered skin of 2 normal birds and 5 from feathered skin of 3 psittacines presented for pathologic feather-picking. All 23 skin specimens consistently adhered to the tape during the biopsy procedure. The specimens were fixed in 10% neutral phosphate-buffered formalin. During processing, no curling or rolling of specimens occurred, and all specimens could be easily orientated for correct trimming and subsequent histopathological evaluation. The tape technique did not produce any appreciable artefacts. Remnants of the tape were microscopically evident above the stratum corneum assuring that none of the stratum corneum was lost during processing. Obtaining avian skin biopsy specimens using this modified tape technique is easy and ensures flat fixation of the skin biopsy specimens, which later allows trimming at right angles, and through the longitudinal diameter of feather follicles for accurate histopathologic evaluation.     

PCR-RFLP和特异PCR技术用于鲁道夫对盲囊线虫鉴定的研究

本研究运用新建立的Contracaecum rudolphii姊妹种的PCR-RFLP和特异PCR鉴定方法对来自我国青海湖的鲁道夫对盲囊线虫进行分子鉴定。经PCR-RFLP分析，来自我国青海湖的鲁道夫对盲囊线虫的两个样品的带型与C．rudolphiiB的酶切带型一致。经特异PCR扩增，C．rudolphii B的特异引物能从我国鲁道夫对盲囊线虫中扩增出目的片段，与对照C．rudolphii B的代表样品扩增片段大小相同。该两种方法试验结果都说明来自我国青海湖的对盲囊线虫为C．rudolphii B，与PCR-SSCP方法鉴定结果一致。本次研究结果及所建立的分子分类学方法为我国异尖线虫的进一步研究奠定了基础。     

Coccidian oocyst and nematode egg counts of free-ranging African buffalo (Syncerus caffer) in the Kruger National Park, South Africa

Faecal specimens collected in the Kruger National Park from 103 African buffaloes (Syncerus caffer) up to 1 year old and 283 buffaloes older than 1 year were examined for the presence of coccidian oocysts and nematode eggs. Most specimens from animals older than 1 year had negative coccidian oocyst counts. Positive specimens from younger animals had significantly higher coccidian oocyst counts than those from older animals. No such difference was found for nematode egg counts.     

Flow cytometric analysis of canine colonic mucosal lymphocytes from endoscopically obtained biopsy specimens

OBJECTIVE: To validate use of canine colonic biopsy specimens obtained via endoscopy as a source of mucosal lymphocytes (ML) for flow cytometric analysis. SAMPLE POPULATION: Mucosal biopsy specimens from 10 adult dogs. PROCEDURE: Mucosal lymphocyte subsets obtained from excised colon were compared with ML subsets obtained from biopsy specimens obtained by use of an endoscopic forceps (6 dogs). Endoscopic colonic biopsy specimens from 4 other dogs were used to define whether obtained ML were predominantly of intraepithelial or lamina propria origin. Mucosal lymphocytes were isolated and labeled, using commercially available monoclonal antibodies directed against canine cell surface antigens. Lymphocyte subsets (cytotoxic or helper T cells; B cells) were determined by use of flow cytometric analysis. RESULTS: A large number of viable ML was obtained after dissociation of the colonic epithelium from excised colon (45.5 + 21.5 X 10(6)) and endoscopic (7.2+/-3.4 X 10(6)) biopsy specimens. Lymphocyte subsets obtained with both methods were identical for each dog and consisted predominantly of intraepithelial lymphocytes, with some lymphocytes from the lamina propria. Collagenase digestion of excised colon also yielded a large number of viable lymphocytes from the lamina propria (56.7+/-20.4 X 10(6)), but collagenase digestion of endoscopic biopsy specimens was less rewarding. CONCLUSION AND CLINICAL RELEVANCE: A representative sample of viable intraepithelial ML is obtainable from endoscopic biopsy specimens. Flow cytometric analysis, a minimally invasive technique, can be used to study ML of client-owned animals.     

Immunocytochemical study of Ki-67 as a prognostic marker in canine mammary neoplasia

Background: Canine mammary tumors are challenging for clinicians and pathologists because of complex histologic classification, low specificity of cytologic diagnosis, and unpredictable biological behavior. In histologic specimens, expression of tumor proliferation marker Ki‐67, a nuclear nonhistone protein, has been shown to have prognostic value for canine mammary tumors and to correlate with malignancy and low survival rates. Objective: The objective of this study was to measure the proliferation index of canine mammary tumors by immunochemical detection of Ki‐67 in cytologic specimens and to determine its relationship to clinical and pathologic variables and patient outcome. Methods: Spontaneous mammary tumors from 31 female dogs were surgically excised. Imprint specimens for cytologic evaluation were wet‐fixed in ethanol; histologic specimens were prepared routinely. Immunostaining was performed with the PH 177 monoclonal antibody against Ki‐67; proliferation index was graded from negative to +++. Dogs were followed for 18 months. Multivariate logistic regression analysis was used to determine correlations between immunocytochemical results, tumor and clinical variables, and patient outcome. Results: Ki‐67 proliferation indices in cytologic specimens were significantly lower for nonmalignant tumors than for malignant tumors. High index values of Ki‐67 were positively correlated with metastasis, death from neoplasia, low disease‐free survival rates, and low overall survival rate. With the exception of 4 specimens for which cellularity was insufficient, positive expression of Ki‐67 in cytologic specimens correlated with that of histologic specimens. Conclusions: The prognostic value of the Ki‐67 index in canine mammary tumors by using wet‐fixed cytology imprint specimens was similar to that observed previously for histologic specimens. Immunocytochemical detection of Ki‐67 could improve the accuracy and value of cytology by providing safe and rapid information about malignancy and patient outcome.     

Stress Protection Afforded by a Cast on Plate Fixation of the Distal Forelimb in the Horse In Vitro

Six forelimb specimens from three adult horses had the fetlock joint fused by application of a dorsal plate and by a screw placed in lag fashion through the metacarpus to each proximal sesamoid bone. Five specimens were instrumented on the central dorsal surface of the plate with a single rosette strain gage, and the plate of the sixth specimen was instrumented with four longitudinally oriented single-axis strain gages. The specimens were loaded axially in compression to 4,000 N in a cast (test 1), in a cast with a heel block (test 2), and uncast (test 3). The principal angle of strain in all specimens, in all tests, closely approximated the vertical axis at loads < 1,000 N. The principal angle in uncast specimens was significantly different at loads > 1,000 N than the cast specimens ( P <.05). At loads > 3,000 N, the principal angle in test 3 closely approximated the horizontal axis, indicating a change from tension to compression on the dorsal surface of the plate, whereas the principal angle of the cast specimens was unchanged. Specimens in a cast (tests 1 and 2) suffered less surface deformation than did uncast specimens (test 3). Therefore, the cast changed the direction and extent of bending at the point of fixation, and thereby decreased the deformation of the plate. This effect would lead to greater fatigue life of the implant in the cast specimens compared with the uncast specimens.     

Analysis of proglycogen and macroglycogen content in muscle biopsy specimens obtained from horses

OBJECTIVE: To determine proglycogen (PG) and macroglycogen (MG) content in equine skeletal muscle and to compare 2 analytical methods (acid hydrolysis [AC] and PG plus MG determination) for measurement of total muscle glycogen content (Gly(tot)) in biopsy specimens. SAMPLE POPULATION: Muscle biopsy specimens obtained from 41 clinically normal horses. PROCEDURE: Forty-five muscle biopsy specimens obtained from the middle gluteal (n = 31) or triceps (14) muscle were analyzed, using AC and MG plus PG determination for Gly(tot). Variability within muscle biopsy specimens for each method was calculated from duplicate analyses of muscle specimens. In a second experiment, variation in MG and PG content between muscle biopsy specimens and the effect of sample collection depth on the concentration of MG and PG in the middle gluteal muscle was evaluated. RESULTS: There was a strong correlation (r = 0.99) between Gly(tot) values obtained by use of AC and MG plus PG determination. Coefficients of variation for within- and between-specimen variability of Gly(tot) were approximately 4% for each method. The PG fraction was always in excess of the MG fraction. Biopsy specimens obtained from the superficial part of the middle gluteal muscle contained significantly more Gly(tot) and PG than specimens obtained from deeper parts. CONCLUSIONS AND CLINICAL RELEVANCE: This study confirms that MG and PG exist in equine skeletal muscle and can be measured reliably in biopsy samples. This technique could be applied in future studies to investigate glycogen metabolism in exercising horses and horses with glycogen-storage diseases.