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.     

Evaluation of hemorrhage, sample size, and collateral damage for five hepatic biopsy methods in dogs

OBJECTIVES: To compare the volume of hemorrhage, number of lobules, and portal triads available for histologic evaluation, and resultant collateral damage between 5 hepatic biopsy methods: biopsy punch, biopsy needle, ligature method, laparoscopic biopsy forceps, and ultrasonically activated scalpel (UAS). STUDY DESIGN: Experimental, repeated measures, block. ANIMALS: Twelve adult dogs. METHODS: Biopsies were obtained from the periphery and center of the left lateral liver lobe of each dog using each of 5 biopsy techniques. Hemorrhage was quantified and compared between methods and sites. Biopsy samples were evaluated histologically to characterize collateral damage and determine the number of lobules and portal triads sampled. RESULTS: Regardless of technique, liver biopsy resulted in minimal hemorrhage (<2 mL). For peripheral biopsies, UAS was comparable with the ligature method, but caused significantly less hemorrhage than all other methods, whereas for central biopsies, UAS caused significantly less hemorrhage than other methods. Except for the laparoscopic biopsy forceps, UAS caused significantly more collateral damage than other methods. UAS and ligature biopsy methods yielded specimens that had more portal triads per sample than other methods. Eight of 48 biopsy needle samples were inadequate for histologic evaluation, whereas other methods yielded adequate specimens. CONCLUSIONS: All biopsy methods produced minimal hemorrhage and except for needle biopsy yielded adequate tissue samples for histologic evaluation. CLINICAL RELEVANCE: Use of UAS is a reliable, safe alternative technique for liver biopsy and can be used laparoscopically to obtain large tissue samples.     

Investigation of Keyes skin biopsy instrument for intestinal biopsy versus a standard biopsy technique

A prospective, clinical trial was conducted using 12 dogs that were presented for intestinal biopsy. Comparisons were made between paired jejunal biopsies collected using a Keyes biopsy punch and a standard scissor excisional technique. There were no differences in speed of collection, diagnostic value of specimens, complication rates, or sample quality. In this study, the Keyes skin punch was found to be safe, rapid, and diagnostic in all animals. Based on results of this study, the use of a Keyes punch instrument can be recommended as an alternative to other open intestinal biopsy methods.     

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.     

Comparison of the effects of the CO2 surgical laser and conventional surgical techniques on healing and wound tensile strength of skin flaps in the dog

OBJECTIVE: To determine if CO(2) laser was superior to conventional surgical techniques (CST) for creation of skin flaps in dogs as determined by hemostasis, wound healing, and wound tensile strength. STUDY DESIGN: In-vivo model ANIMALS: Six large, mixed-breed dogs. METHODS: On each dog's trunk, 3 pairs of identical, dorsally based, pedicled skin flaps were created and sutured back into position. Based on a Latin Square design, flaps on one side were created with a CO(2) laser and on the other side by CST. Intraoperative hemorrhage was measured by weighing sponges used to absorb blood. On days 7, 10, and 14, specimens from the flap-skin junction and the central portion of each pair of flaps were collected. Specimens were subjected to tensile strength testing and histologic examination to evaluate wound healing. RESULTS: Mean (+/- standard error of mean [SEM]) hemorrhage was significantly less (P =.02) with CO(2) laser (4.70 +/- 1.37 g) than CST (10.82 +/- 1.37 g). Wound tensile strength (N/m(2)) after CST (0.49 +/- 0.049) was significantly greater (P =.01) than with CO(2) laser (0.17 +/- 0.049). Laser incisions had partial necrosis of the wound edges and a more extensive inflammatory response; however, healing of the wound beds were similar regardless of technique. CONCLUSIONS: The CO(2) laser provided better intraoperative hemostasis than CST. However, overall healing and increase in tensile strength of the skin-flap junction of the flaps created by the laser may be delayed during the first few weeks of wound healing. CLINICAL RELEVANCE: Skin flaps used to repair large skin defects in dogs can be created and elevated with a CO(2) laser. In areas of increased skin mobility or tension, skin flaps created with CO(2) laser may be more susceptible to complications such as dehiscence, and care should be taken to minimize these complications.     

The effect of ketoconazole on whole blood and skin ciclosporin concentrations in dogs

Background –  Ciclosporin (CSA) is approved for the treatment of canine atopic dermatitis. Ciclosporin is metabolized by liver cytochrome P450 enzymes, a process inhibited by ketoconazole (KTZ). Hypothesis/Objectives –  The aims of this study were to determine skin and blood CSA concentrations when CSA was administered alone at 5.0 (Treatment 1) or 2.5 mg/kg (Treatment 2) and when CSA was administered at 2.5 mg/kg concurrently with KTZ at 5 (Treatment 3) or 2.5 mg/kg (Treatment 4). We hypothesized that skin and blood CSA concentrations in Treatment 1 would not differ from those obtained with T3 or T4. Animals –  In a randomized cross‐over study, six healthy research dogs received each of the treatments (Treatment 1, 2, 3 and 4) once daily for 7 days. Methods –  After the first, fourth and seventh dose for each treatment, a peak and trough skin punch biopsy sample and whole blood sample were collected and analysed with high‐performance liquid chromatography–tandem mass spectrometry. Data were analysed using a repeated measures approach with PROC MIXED in SAS. Pairwise comparisons were performed with least squares means and Tukey–Kramer adjustment for multiple comparisons. Results –  Mean blood CSA concentrations in Treatment 1 were not different from those in Treatment 2 or 4, but were less than in Treatment 3. Mean skin CSA concentrations in Treatment 1 were greater than in Treatment 2, not different from those in Treatment 4, and less than those in Treatment 3. Conclusions and clinical importance –  Administration of CSA and KTZ concurrently at 2.5 mg/kg each may be as effective as CSA alone at 5.0 mg/kg for treatment of canine atopic dermatitis.     

Diagnostic accuracy of pre‐treatment biopsy for grading soft tissue sarcomas in dogs

Histologic grade is an important prognostic factor for both local recurrence and metastatic potential with canine soft tissue sarcoma (STS). Pre‐treatment biopsy with identification of tumour grade may aid in prognostication and determination of surgical margins necessary for local control. The purpose of this study was to evaluate the grading accuracy of various pre‐treatment biopsy techniques (wedge, punch, needle‐core) for STS in dogs. Medical records of 68 dogs diagnosed with a STS via pre‐treatment biopsy and confirmed by excisional biopsy were evaluated. The concordance in grade between excisional and pre‐treatment biopsies was 59%. Of the 41% that lacked concordance, 29% of pre‐treatment biopsies underestimated and 12% overestimated grade. The method of pre‐treatment biopsy did not significantly effect grade concordance. Based on these data, needle‐core biopsy appears to be similar in accuracy compared to open biopsy, however, grading determined by pre‐treatment biopsy in general should be interpreted with caution.     

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.     

Immunoglobulin-E-bearing cells in skin biopsies of horses with insect bite hypersensitivity.

The aim of the present study was to investigate, with immunohistochemistry and in situ hybridisation, if immunoglobulin-E (IgE) and mast cells are involved in the pathogenesis of insect bite hypersensitivity (IBH), an allergic dermatitis of horses. In tissue sections fixed in paraformaldehyde (PFA) for <24 h, significantly more IgE protein-bearing cells were found in the dermis and epidermis of acute and chronic IBH lesions than in skin biopsies from healthy horses (medians = 466, 236 and 110 cells/mm2, respectively; P < or = 0.01). More IgE-mRNA positive (+) cells were observed in the dermis of acute IBH lesions than in the dermis of healthy skin (median = 2.8 vs. 0.0 cells/mm2; P < or = 0.01). Significantly, more mast cells were detected with metachromatic (median = 160 vs. 62 cells/mm2; P < or = 0.001) and tryptase-specific stainings (median = 120 vs. 69 cells/mm2; P < or = 0.001) in the dermis of acute IBH biopsies compared to healthy skin. No chymase+ mast cells were found in any skin biopsy. IBH lesions fixed in PFA for >24 h were compared to dermatomycosis (DM) lesions; IBH biopsies contained a similar number of IgE-protein+ cells to DM biopsies (median = 249 vs. 192 cells/mm2; P = 0.08) but had significantly more IgE-mRNA+, metachromatic and tryptase+ mast cells than DM biopsies. This study suggests an involvement of IgE-mediated immune reactions in the pathogenesis of IBH as well as, sometimes, in dermatomycosis. Using double labelling, cells which expressed IgE protein and contained mast cell enzymes were detected.     

COMPARISON OF GLOMERULAR NUMBER AND SPECIMEN LENGTH OBTAINED FROM 100 DOGS VIA PERCUTANEOUS ECHO-ASSISTED RENAL BIOPSY USING TWO DIFFERENT NEEDLES

Our objective was to evaluate possible differences in the number of glomeruli and length of renal biopsies collected in canine subjects by two different types of biopsy needles: a semiautomatic 18-gauge Trucut and an automated 18-gauge Jamshidi modified (Biopince). One hundred biopsy samples obtained from dogs of different ages and gender affected by different nephropathies were evaluated retrospectively. All animals were biopsied using one of the two different needles. Biopsies were performed under ultrasound guidance and evaluated by a single pathologist. Statistical analysis was performed to evaluate possible differences in the number of glomeruli and length of renal biopsies collected comparisons were determined between subgroups of dogs with or without the identification of renal interstitial infiltrates and/or fibrosis. Neither the mean difference of the number of glomeruli nor the length of tissue sample collected with the different needles was significantly different. Likewise, the average biopsy length did not differ in dogs with or without renal interstitial infiltrate in animals biopsied with either biopsy needle. Both the Biopince and the Trucut devices provide diagnostically adequate biopsy renal specimens using ultrasound-guidance.     

High-frequency ultrasonography of the skin of clinically normal dogs

OBJECTIVE: To assess the applicability of high-frequency diagnostic ultrasonography for evaluation and accurate measurement of the skin thickness of clinically normal dogs. ANIMALS: 26 healthy dogs (12 sexually intact males, 13 sexually intact females, and 1 spayed female) of various breeds and ages. PROCEDURE: Ultrasonographic examination of the skin and histomorphometric analysis of skin biopsy specimens obtained from the same site were performed. A 13-MHz linear-array transducer was used to obtain a series of ultrasonographic images of the skin in the flank region; images were analyzed and measured by use of imaging software. Cutaneous biopsy specimens were placed in fixative and then stained with H&E and Masson trichrome stains. Histomorphometric analysis was performed by use of an image analyzer. Thickness of the epidermis and dermis of each specimen was evaluated by use of a semiautomatic procedure of quantification. Data obtained from ultrasonographic and histologic measurements were compared by use of the Pearson correlation test. RESULTS: The ultrasonographic pattern of canine skin was consistently characterized by 3 distinct, defined echogenic layers corresponding to the epidermal entry echo, epidermis and dermis, and subcutaneous tissues. A positive correlation was found between ultrasonographic and histologic measurements of skin thickness. CONCLUSIONS AND CLINICAL RELEVANCE: Comparison between ultrasonographic and histologic appearance of the skin revealed that layering of canine skin (ie, epidermis and dermis) and the subcutaneous tissues may be recognized and measured by use of high-frequency ultrasonography. Thus, diagnostic ultrasonography may be a useful tool for the noninvasive evaluation of cutaneous disorders in dogs.     

The effects of surface preparation on the penetration of hydrocortisone through canine skin

This study investigated the effects of common skin surface preparations on the penetration kinetics of hydrocortisone through canine skin. Thoracic skin from five dogs was clipped of hair, divided between five treatment groups and prepared as follows: shaved (S); tape-stripped with adhesive bandage (TS); cleaned with aqueous chlorhexidine (Aq-C); cleaned with alcoholic chlorhexidine (Al-C); or allocated to the control group and had no further preparation performed (C). The skin samples were mounted in Franz-type diffusion cells and transdermal hydrocortisone penetration was measured over 30h. The pseudo-steady-state flux (J(SS)) of hydrocortisone through S, Al-C, Aq-C and TS skin was, respectively, 2.3 (P=0.021), 2.2 (P=0.037), 2.0 (P=0.070) and 1.5 (P=0.351) times greater than through the control skin, but there were no significant differences in the lag times (t(lag)) for hydrocortisone penetration between the groups. The study has shown that some skin surface preparations can significantly increase the subsequent penetration of hydrocortisone through canine skin in vitro.     

Defocused CO2 laser on equine skin: a histological examination

REASONS FOR PERFORMING STUDY: No studies have been published on effects of treatment with a defocused beam carbon dioxide (CO2) laser on equine skin histology. A better understanding of this will help to define how lasers should be used, in order to reduce potential side effects. OBJECTIVE: To describe the acute effects of different doses of defocused CO2 laser, ranging from therapeutic to surgical levels, on equine skin. METHODS: Defocused CO2 laser was administered to the skin in the hamstrings (91 J/cm2), fetlock (137 J/cm2) and loin (450 J/cm2) areas of 13 Standardbred horses. The acute effects on skin histology were examined 90 min after the end of therapy. RESULTS: Mild changes with focal spongiosis and subepidermal clefts were found after 91 J/cm2 irradiation and more severe changes with diffuse subepidermal clefts after the 137 J/cm2 dose. A homogeneous eosinophilic acellular zone of dermis and destruction of adnexal structures, and significant thinning of the epidermis was observed after the 450 J/cm2 dose. CONCLUSIONS: The present study indicates acute dose-dependent changes in equine skin histology after laser treatment Severe tissue damage was induced using a 450 J/cm2 dose. POTENTIAL RELEVANCE: To reduce the potential side effects of defocused CO2 laser treatment, the laser parameters must be carefully evaluated. Caution should be taken if doses higher than 91 J/cm2 (16 W, 4 min, and 42 cm2) are used in irradiation of equine skin.     

Thermal injury by in vitro incision of equine skin with electrosurgery, radiosurgery, and a carbon dioxide laser.

Freshly harvested equine skin incised with an electrosurgical unit, a radiosurgical device, or a carbon dioxide (CO2) laser was examined by light microscopy to determine the extent of thermal injury caused by each instrument. There was no significant difference between the thermal injury caused by the electrosurgical unit in the pure-cut mode and the CO2 laser in the superpulse mode, or between the electrosurgical unit and the radiosurgical device in the fully filtered cut mode. However, thermal injury caused by the CO2 laser was significantly less than that caused by the radiosurgical device. The amount of thermal injury in this in vitro study was similar to that found in vivo with other species.     

A NEW DEVICE FOR STEREOTACTIC CT-GUIDED BIOPSY OF THE CANINE BRAIN: DESIGN, CONSTRUCTION, AND NEEDLE PLACEMENT ACCURACY

An inexpensive device was created for computed tomographic (CT)-guided stereotactic biopsy of the canine brain. The accuracy of the device was tested using 16, formalin-perfused, canine head specimens. For each dog, a 6-inch biopsy needle was guided into pituitary gland and caudate nucleus targets. Needle tracks were measured using the CT computer and infused with tissue staining solution. Hit success and actual needle track lengths were determined from sliced brain specimens. The device enabled accurate orientation and placement of the canine head in the slice plane, such that progressive penetration of the biopsy needle could be monitored. The caudate nucleus was hit 12/16 times (75% accuracy) and the pituitary gland 15.5/16 times (98.6% accuracy). Hit proportions for the two targets did not differ (P < 0.05). A significant difference was found between CT and actual track length for both targets (P < 0.01). This was attributed to incomplete staining of the bevel portion of the needle track.     

Sulphido-leukotriene production from peripheral leukocytes and skin in clinically normal dogs and house dust mite positive atopic dogs

Pathogenesis of canine atopy has not been completely elucidated. In humans, sulphido-leukotrienes (s-LT) play a role in atopy, and increased production of s-LT occurs in the skin and peripheral leukocytes after allergen challenge. The study population included 16 clinically normal and 13 atopic dogs. All atopic dogs had in common a positive reaction (4+) to the intradermal injection of house dust mite (allergen of reference). Blood samples and skin biopsies were collected. Sulphido-LT synthesis by peripheral leukocytes after stimulation was measured, and no statistically significant difference was found between clinically normal and atopic dogs. Sulphido-LT concentrations in skin samples from stimulated and unstimulated sites were measured, and no statistically significant difference was detected between clinically normal and atopic dogs or between lesional and nonlesional skin within the atopic group. Clinical signs of atopic dogs were graded by owners and no correlation was found between their severity and cutaneous concentrations of s-LT. In this study there was no increase in s-LT synthesis in atopic dogs.     

Thermal injury and wound healing of the endometrium subsequent to minimally invasive transendoscopic use of Nd:YAG-laser-and electrosurgery in horses

Transendoscopic thermal preparation techniques like laser- or electrosurgery have proved to be most reliable for minimal invasive intrauterine surgery during operative hysteroscopy in mares. To determine the effect of Nd:YAG laser surgery on the endometrium and the complete uterine wall compared with electrosurgery, standard lesions were obtained by transendoscopic monopolar electrosurgery (loop electrode, cutting blade) and Nd:YAG laser surgery (contact, bare fibre, 25 Watt, exposure time four seconds, non contact, 80 Watt, exposure time four seconds) in five healthy mares on days d 0, 7, 14, 18 and 21. The gross effects were controlled endoscopically before the consecutive lesions were set. Following the last diathermic endometrial irritation, hysterectomy was performed under general anaesthesia. Recovery of the mares was uneventful. Specimens from the lesions were collected after dis-section of the uterus, fixed in formalin, routinely embedded and stained with H. E. for histological evaluation. The different energy forms caused tissue alterations of comparable quality but differing in degree. Most intensive peripheral hyperemia and tissue edema occurred on postoperative days three to seven. Non contact laser irradiation initiated maximal amount of fibrinous exudate. Lesions created with a loop electrode were superficial only and did only affect the endometrium. Preparation with an electrosurgical cutting blade or a laser bare fibre resulted in craterlike lesions lined by carbonisation and did involve deeper layers of the myometrium. The lesions caused by non contact laser irradiation affected all layers of the uterine wall. Three weeks postoperatively, reepithelization of the luminal epithelium was completed. The depth of thermal injury and coagulative necrosis, inflammatory reactions and scar formation was greater in lesions created by laser application compared with those set with a loop electrode at power densities comparable to those usually achieved at hysteroscopic surgery. The results suggest that an electrosurgical loop designed for operative hysteroscopy may be a reliable tool for diathermic resection of pathological endometrial structures like uterine cysts characterized by minimal thermal injuries and a short period of reconvalescence. Non contact laser irradiation may result in deep thermal injuries with the risk of delayed uterine perforation but may be profitable for treatment of partly intramural structures like an intramural leiomyoma.     

Effect of solute lipophilicity on penetration through canine skin

OBJECTIVE: To investigate the effect of lipophilicity on the percutaneous penetration of a homologous series of alcohols through canine skin. DESIGN: Skin harvested from Greyhound thorax was placed in Franz-type diffusion cells and the in vitro passage of radiolabelled (14C) alcohols (ethanol, butanol, hexanol and octanol (Log P 0.19-3.0)) through separate skin sections was measured in replicates of five. Permeability coefficient (kP, cm/h), maximum flux (Jmax, mol/cm2/h) and residue remaining within the skin were determined. RESULTS: The kP increased with increasing lipophilicity (6.2 x 10(-4) +/- 1.6 x 10(-4) cm/h for ethanol to 1.8 x 10(-2) +/- 3.6 x 10(-3) cm/h for octanol). Alcohol residues remaining within each skin sample followed a similar pattern. An exponential decrease in Jmax with increasing lipophilicity was observed. CONCLUSION: Changes in canine skin permeability occur with increasing alcohol lipophilicity. This finding has practical consequences for the design of topical formulations and optimisation of drug delivery through animal skin.     

Evaluation of two commercial enzyme-linked immunosorbent assays for detection of bovine viral diarrhoea virus in serum and skin biopsies of cattle

AIM: To assess the ability of two commercial bovine viral diarrhoea (BVD) virus (BVDV) antigen-capture enzyme-linked immunosorbent assays (ELISAs) to detect virus in serum and skin biopsies. METHODS: Thirty cattle persistently infected (PI) with BVDV were identified using routine diagnostic laboratory testing. Additional ear-notch skin biopsies and blood samples were collected from these animals to confirm the diagnosis, and from 246 cohorts, to determine their BVDV status. Skin biopsies were soaked overnight in buffer and the eluate collected. All sera and eluate were tested using two commercially available ELISAs for detecting BVDV antigen, and a subsample of positive and negative sera was tested using a polymerase chain reaction (PCR) test. A study was also performed to ascertain the risk of cross contamination occurring during the collection and processing of skin biopsies. RESULTS: Both serum and skin samples tested using either ELISA resulted in the detection of all cattle identified as PI and no non-infected cattle were incorrectly classified as infected using either method. Agreement between all assays (ELISAs, whether performed on serum or skin, and PCR) was 100%. No cross-contamination of skin samples between animals was evident using routine biopsy methods. CONCLUSIONS: Viraemic cattle infected with BVDV were accurately identified using either of the two commercial ELISAs evaluated on either serum or skin samples. CLINICAL RELEVANCE: Either skin biopsies or serum samples can be collected from cattle to determine their BVDV status. This should overcome problems in accurately identifying the infection status of young calves in which colostral antibodies might interfere with the antigen-capture ELISA.     

Expression analysis of CCL27 and CCL28 mRNA in lesional and non-lesional skin of dogs with atopic dermatitis

Chemokines are important regulators of the selective recruitment of inflammatory cells into sites of allergic inflammation. Since canine atopic dermatitis (AD) shares many clinical features of human AD, patterns of chemokine production in dogs may also be similar with those in humans. The aim of this study was to examine mRNA expression of CCL27 and CCL28 in lesional skin of dogs with AD to demonstrate similarity of chemokine production with human counterparts. RNA was extracted from skin biopsy specimens of 12 dogs with AD. The mRNA expression of CC chemokines (CCL4, CCL19, CCL20, CCL21, CCL24, CCL27 and CCL28) was analyzed by quantitative real-time PCR and was compared between lesional and non-lesional skin. Seven types of chemokines examined were constitutively expressed in both lesional and non-lesional skin. It was found that mRNA expression levels of CCL27 and CCL28 among the chemokines were significantly different between lesional and non-lesional skin (P<0.05). Expression level of CCL27 mRNA in lesional skin was significantly lower than that in non-lesional skin. On the other hand, CCL28 mRNA expression in lesional skin was found to be higher than that in non-lesional skin. These results suggest that CCL28 but not CCL27 may play important roles in immunopathogenesis of canine AD, indicating that experimental canine study may provide additional information that can be extrapolated to human AD.