The distribution and heterogeneity of mast cells in tongue from five different avian species

This study was conducted with the aim of determining the morphology, distribution and heterogeneity of mast cells in the tongues of seagull (Larus fuscus), common buzzard (Buteo buteo), goose (Anser anser), white stork (Ciconia ciconia) and Gerze rooster. The study used five samples of tongue material from each of the healthy adult avian species. The samples were fixed in 10% neutral‐buffered formalin (NBF) solution, then, after routine tissue follow‐up, the samples blocked with paraplast. Cross‐sections with 5–6 μm of thickness were stained with the 0.5% toluidine blue and alcian blue/safranin O (AB/SO). In all five avian species, it was found that the mast cells were in different sizes and round, oval or spindle‐shaped based on their place of distribution. Mast cell numbers were determined in stained with toluidine blue, examined ×40 objectives in a 1 mm² area. It was observed that mast cell density in subepithelial lamina propria and microscopic papilla was higher in the tongues of all species. Mast cell distribution and heterogeneity varied through the tongue, and there were more mast cells in the dorsal side of the tongue than the ventral side. The highest amount of mast cells was found in the tongue of the Gerze rooster among all five species. In the tongue cross‐sections stained with the combined method of alcian blue/safranin O (AB/SO), the mast cells were stained as AB (+), SO (+) and AB/SO (+) (mixed).     

Variation in gross and histological appearance of the canine pars flaccida

The tympanic membrane is divided into the pars flaccida (PF) and pars tensa. Otoscopic differences in the appearance of the PF in the dog (flat vs. bulging) have been reported anecdotally. The purpose of this study was to histologically examine the canine PF to determine if differences exist between a flat and a bulging PF. We hypothesized that differences in the PF may be structural or due to an increased pressure in the middle ear. Four adult canine cadavers were used (two had bilateral bulging PF, two had bilateral flat PF). The ear specimens were fixed in 10% buffered neutral formalin, decalcified, routinely processed and stained with haematoxylin and eosin, toluidine blue (staining for mast cells), and Verhoeff-van Gieson (staining for elastic fibres). One blinded investigator evaluated each PF histologically. Seven ears (three with bulging PF, four with flat PF) were evaluated in the study. The PF was identified in all seven ears. No histological differences that distinguished a bulging PF from a flat PF were identified. Six ears were evaluated for the presence of mast cells and elastic fibres. Less than one mast cell per x40 field was identified in the PF in five of six ears. No elastic fibres were identified in any of the six PF. Based on the results of this study, it appears unlikely that there is a structural difference that accounts for a bulging PF. Therefore, there may be increased middle ear pressure in dogs with a bulging PF.     

绒山羊肥大细胞类胰蛋白酶的免疫组化及图像分析研究

[目的]探讨绒山羊肥大细胞中是否存在类胰蛋白酶及不同固定液对其常规染色和免疫组化染色结果的影响。[方法]采用兔抗羊肥大细胞类胰蛋白酶多克隆抗体间接免疫过氧化物酶技术检测由Carnoy液和中性福尔马林液(NBF)固定的绒山羊空肠、瓣胃和肺等组织肥大细胞中是否存在类胰蛋白酶。同时采用Image-ProPlus图像分析软件和人工计数法对结果进行分析,比较经常规染色和免疫组化染色后不同固定液固定组织中肥大细胞的形态及数量,进而判断Carnoy液和中性福尔马林液(NBF)对该检测技术的影响。[结果]兔抗羊多克隆抗体与绒山羊肥大细胞中的类胰蛋白酶具有良好的交叉反应,证实绒山羊肥大细胞胞浆颗粒中存在类胰蛋白酶。同时,与Carnoy液相比较,NBF液固定组织能较好地反映肥大细胞在组织中的数量和形态变化。[结论]绒山羊肥大细胞中存在类胰蛋白酶,且与Carnoy液相比较,NBF液是一种更为适合绒山羊肥大细胞常规染色和免疫组化染色的固定剂。     

Comparison of tracheal aspirates and bronchoalveolar lavage in racehorses. 1. Evaluation of cytological stains and the percentage of mast cells and eosinophils

OBJECTIVE: To compare a fast Romanowsky cytological stain (Diff-Quik) and Leishman's stain for the detection of mast cells in samples from the lower airways of racehorses, and to compare the proportion of mast cells and eosinophils in the total inflammatory cells in tracheal aspirate (TA) with those in paired bronchoalveolar lavage (BAL) samples. DESIGN: Retrospective case series of 48 young Thoroughbred and Standardbred racehorses. PROCEDURE: Fifty-one paired TA and BAL samples were collected after treadmill exercise from 48 horses with poor racing performance. Two slides were prepared from each sample; one was stained with Diff-Quik stain and the other with Leishman's stain. Differential cell counts of eosinophils and mast cells were recorded from each slide. Comparison of the suitability of the stains for the detection of mast cells, and comparisons of eosinophil and mast cell percentages in TA and BAL samples were analysed using the non-parametric Wilcoxon matched pairs test. RESULTS: Percentages of mast cells were significantly higher in Leishman than in Diff-Quik stained slides in both TA (P = 0.03) and BAL samples (P < 0.0001). Mast cell percentages were significantly higher in BAL than in TA samples using Leishman's stain (P < 0.0001). There was no significant difference in eosinophil percentages between TA and BAL samples (P = 0.07). CONCLUSIONS: Fast Romanowsky type stains (for example Diff-Quik) are not appropriate for the detection of mast cells in samples from the equine lower respiratory tract. Therefore, a metachromatic stain that reliably identifies mast cells (for example Leishman's) should be used if evaluation of mast cells in lower respiratory tract is undertaken. Mast cells are predominantly found in the distal small airways and alveoli sampled with a BAL. In contrast, eosinophils appear to be evenly distributed in the lower respiratory tract. However, high percentages of eosinophils are occasionally found only in TA samples. We recommend that both a TA and BAL be used for the evaluation of eosinophils and mast cells within the equine lower respiratory tract.     

Stereology of the liver in three species of Leontopithecus (Lesson, 1840) Callitrichidae--primates

Studies on liver morphology and stereology are relevant to the comparative anatomical and pathological research. They also facilitate the use of non-human primates in basic research, which has substantially supported studies in human medicine. Quantitative studies of liver structures have also been more extensive in Old World primates and other vertebrates. Twenty-three livers of adult lion tamarins were studied (six Leontopithecus rosalia, seven Leontopithecus chrysomelas, and 10 Leontopithecus chrysopygus), dissected, and fixed in 10% neutral buffered formalin solution. For stereological quantification, the liver was regarded as consisting of parenchyma (hepatocytes) and stroma (non-hepatocytes). The volume density (V(v)) was determined by point counting, and the disector method was used to obtain the numerical density of hepatocytes (N(v)). Hepatic stereological differences among the three species of lion tamarins were not statistically significant. Therefore, the pooled V(v[hepatocyte]) and V(v[stroma]) could be determined as 96.2 and 7.4%, respectively, and N(v[hepatocyte]) as 500.33 x 10(6) cm(-3). Significantly different, the values found for V(v[hepatocyte]) and N(v[hepatocyte]) in lion tamarins were, respectively, 0.09 and 2.8 times greater than those in baboons, and 0.17 and 3.8 times greater than those in man. However, the V(v[stroma]) was 1.04 times smaller than that in baboons and 1.79 times smaller than that in man.     

Diarrhea associated with intestinal Cryptosporidiosis in turkeys

Turkey poults were suffering from diarrhea on a farm in which several previous grow-outs of turkeys had experienced a clinically identical problem. Upon necropsy, significant gross lesions were restricted to the gastrointestinal tract. Segments of small intestine were pale and distended with cloudy mucoid material and a few gas bubbles. The ceca contained fluid and gas. Fresh organ portions were collected and fixed in 10% neutral buffered formalin and submitted for histological processing and examination (light microscopy and scanning and transmission electron microscopy). Small (2-4 micron) basophilic bodies identified as Cryptosporidium sp. were present in enterocytes of the middle and lower small intestine. The villi were moderately atrophic, the crypts were hypertrophic, and the lamina propria was infiltrated by large numbers of lymphocytes, heterophils, and fewer macrophages and plasma cells. Numerous intraepithelial leukocytes and exocytosing inflammatory cells also were present.     

Distribution and ultrastructure of mast cells in the equine respiratory tract

Mast cells in the equine respiratory mucosa were studied at both light--and transmission electron--microscope levels. Mast cells were identified at all levels of the tract, with the greatest cell density in the nasopharynx. The majority (57 to 94 per cent) of this cell population were located within the connective tissue of the lamina propria. Up to 20 per cent of these cells were associated with the mucosal glandular tissue, whilst small numbers were present within the surface epithelium and in association with nodular lymphoid tissue. In the peripheral lung tissue 20 per cent of the mast cell population was associated with the small airways, 25 per cent with the pleura, and 32 per cent with blood vessels. The fine and ultrastructural features of these mast cells resemble those described in other species, and their granules consist of two types which resemble human mast cells.     

Distribution of eosinophil granulocytes and mast cells in the reproductive tract of female goats in the preimplantation phase

Changes in eosinophil granulocytes and mast cells post-insemination may affect conceptus implantation, but information regarding the numbers of such cells in the mammalian reproductive tract is limited. This study investigated the preimplantation distribution of eosinophil granulocytes and mast cells (MCs) in the reproductive tract organs of female goats. Uterus, uterine cervix and uterine tubes samples were obtained at slaughter. Cornu uteri were washed in phosphate buffer solution (each animal contained at least one embryo). Tissues were fixed in 10% neutral buffered formol, Carnoy solution and Mota’s fixative (basic lead acetate) for 48 h and embedded in paraffin. Six-micrometre-thick sections were stained with Congo red (for eosinophil granulocytes) and toluidine blue in 1% aqueous solution at pH 1.0 for 5 min (for MCs). In the uterus, MCs occurred in highest numbers in the myometrium. Higher MC numbers were observed in uterus, uterine and uterine tubes in the preimplantation (experimental) group (cycle synchronised through 7 days intravaginal sponge with 0.3 g P₄) compared with the control group (P < 0.05). Eosinophil granulocyte numbers were significantly higher in the experimental group than in the control group (P < 0.05). These results indicate preimplantation-related changes in numbers of eosinophil granulocytes and MCs in goat reproductive tract organs.     

Cow breeding and calf growth performance as affected by pregnancy status the previous year

A 2-yr study was conducted to compare the subsequent cow breeding and calf performance of cows that were nonpregnant with cows that were pregnant at the time calves were weaned. Cows were Angus (A), Polled Hereford (PH), Santa Gertrudis (SG) straightbreds and crossbreds of these breeds. Nonpregnant cows (G1) were 4- to 9-yr-olds that had a calf the previous year and appeared to be physically sound with no detection (by rectal palpation) of an abnormal reproductive tract due to disease, abnormal growth or calving difficulties. Pregnant cows (G2) were of similar age and breed composition to G1 cows. The 93 G1 and the 193 G2 cows were assigned within age and breed composition to sire breeding groups on pasture in an approximate 1:2 ratio, respectively, per sire. There were six A, three PH and one SG sires. The year prior to G1 cows being nonpregnant, G1 cows calved 11 d later (P less than .01) than G2 cows. Subsequent to their being nonpregnant, G1 cows gained 27 kg more (P less than .001) weight during the breeding period, had 5.4 percentage units more (P less than .29) calves born, had calves 17 d earlier (P less than .001) in the calving period, had calves that gained at a similar rate to weaning and had calves that were 14 kg heavier (P less than .01) at weaning (due to their being 17 d older) compared with G2 cows and calves.     

Use of trainer animals to improve performance and health of newly arrived feedlot calves

Four trials were conducted to determine the efficacy of using trainer animals to improve the health and performance of newly arrived feedlot calves. For all trials, trainer animals were given 3 wk to adapt to the feedlot before arrival of the feeder calves and initiation of the trials. Trainer animals were present with newly received feedlot calves for 14 d after arrival and then were removed from the pens for the remaining 14 d of the experiments. In Trial 1, trainer animals were six crossbred beef steers and six mature cull beef cows. Newly received calves were allotted to 18 pens with 10 calves/pen. Six pens contained a trainer steer and six pens contained a trainer cow. Similar procedures were used for the subsequent three trials, except 12 trainer cows and 24 pens were used, and in Trial 4 half of the calves were allotted to pasture paddocks for 14 d before placement in their feedlot pens. During wk 1 of Trial 1, calves with trainer cows and steers gained weight more rapidly (P < .10) than those without a trainer animal (1.12 vs .67 kg/d, respectively). During wk 2, this trend was reversed and overall gains did not differ (P > .20) among treatment groups. Morbidity was 16.7 for control calves, 28.3% for calves with trainer steers, and 8.3% for calves with trainer cows. Four of six trainer steers required antibiotic treatment for respiratory disease. On d 1, a greater (P < .05) percentage of calves in the trainer cow group (81.7%) were observed eating during the first 30 min after feeding compared with either the steer trainer group (60%) or the control group containing no trainer animal (48.3%). This trend continued on d 2 but was not evident on d 3 or 7. In Trial 2, overall gains were 10% greater (P < .06) and final BW was higher (P < .01) for calves with trainer cows than for those without trainers. Trainer cows resulted in a substantial reduction (P < .01) in calf morbidity compared with calves housed alone. In Trial 3, trainer cows did not improve performance or health of newly received calves. More (P < .07) calves with trainers than without were eating 5 min after feeding on d 1, 2, 4, and 8. In Trial 4, the presence of trainer cows the first 2 wk did not affect (P > .27) gains. However, calves placed on pasture after arrival had lower (P < .03) gains during wk 1 than those housed in the feedlot. Calves placed in pasture paddocks upon arrival had more than twice (P < .01) the incidence of morbidity of those placed directly in the feedlot. In these trials, trainer cows had a significant effect on eating behavior of newly received calves, but health and performance benefits were variable.     

Detection of Ehrlichia risticii using an avidin-biotin-immunoperoxidase staining system

An indirect immunoperoxidase procedure using a specific anti-Ehrlichia risticii monoclonal antibody and an avidin-biotin-peroxidase staining method was used to detect E. risticii antigen in infected P388D1 murine monocytes. Several different methods of cytological fixation were used, including acetone (15 min), 95% ethanol (15 min), Bouin's fixative (5 hr), and 10% buffered neutral formalin (24 hr). The E. risticii organisms were labeled effectively and identified in cells fixed with acetone and ethanol. However, infected P388D1 cells fixed in 10% formalin or Bouin's fixative required enzymatic digestion with 1.0% trypsin for 15 min at 37 C before positive results were evident. This indirect immunoperoxidase avidin-biotin staining procedure proved to be a sensitive assay for the detection of intracellular E. risticii and may be an effective diagnostic procedure for formalin-fixed paraffin-embedded tissue.     

Changes of vitamin E status of periparturient dairy cows and newborn calves

Eleven primiparous Holstein Friesian and their crossbred calves (F1, Japanese Black cattle × Holstein Friesian) and 10 multiparous Holstein Friesian and their Holstein Friesian calves were used to evaluate vitamin E status in periparturient period. Plasma α‐tocopherol (α‐toc) concentrations of the multiparous cows were significantly higher than those of the primiparous cows from 60 days before expected calving to 90 days of lactation (P < 0.05). The multiparous cows had a further decrease in the concentrations of α‐toc and total lipid in plasma to the calving than the primiparous cows. Colostrum α‐toc concentrations in multiparous cows were significantly higher than those of the primiparous cows (P < 0.05). Plasma α‐toc concentrations of calves borne by the multiparous cows were significantly higher than those of the primiparous cows at 5 days of age (P < 0.05). Plasma α‐toc concentrations of calves were highest at 5 and 15 days of age in the calves borne by the multiparous and primiparous cows, respectively, and decreased thereafter till 90 days of age. The higher vitamin E status of multiparous cows over primiparous cows might have reflected nutritional composition in the rations. Their calves afforded higher plasma α‐toc levels after birth because of more α‐toc transfer via placenta and more α‐toc secretion in the colostrums thereafter. Plasma α‐toc concentrations of the calves might have decreased as the calves became dependent upon the solid feed of low vitamin E content.     

Mast cells of the bovine trachea: staining characteristics, dispersion techniques and response to secretagogues.

Sections of the lower trachea of cattle, fixed in either Carnoy''s or formalin, were stained with toluidine blue, alcian blue, or alcian blue and safranin O to study the mast cell population. After toluidine blue staining, about twice as many cells in tissue fixed in Carnoy''s contained dark blue granules compared with tissue fixed in formalin. In addition, for the first time in cattle, a population of cells containing red granules was identified after staining with alcian blue and safranin O. Most of these red granules were formalin sensitive. An enzymatic dispersal technique for mast cells is described that yielded 9.4+/-0.4% mast cells (percentage of nucleated cells) with a viability of 92.3+/-0.6%. Spontaneous histamine release was 3.3+/-0.8%. Dispersed mast cells were challenged with various immunological and nonimmunological secretagogues. The calcium ionophores, A23187, ionomyocin, and BrX537A, were effective in releasing up to 94% of histamine in mast cells in a dose-response relationship. Pasteurella haemolytica culture supernate caused about 10% histamine release at a dose of 0.5 mg/mL after correction for spontaneous release. The average histamine content of the mast cells was 6.6+/-1.0 pg/cell. Cytospins of dispersed cells fixed in Carnoy''s and stained with alcian blue and safranin O contained mast cells with blue and red granules, and a few cells with a mixture of both granule types. Based on the effects of type of fixation, staining characteristics and histamine content, a mix of subtypes of mast cells is present in the bovine trachea. However, functionally they respond to secretagogues differently than rodent mast cells. Without an immunological secretagogue, studies to determine compounds that will be effective in blocking mast cell degranulation will be limited.     

Types of myofibers parasitized in experimentally induced infections with Sarcocystis cruzi and Sarcocystis capracanis

Eighteen calves were orally inoculated with either 200,000 or 225,000 sporocysts of Sarcocystis cruzi. Eight goats were orally inoculated with 20,000 sporocysts of S capracanis. Calves and goats were euthanatized at various times after inoculation, and portions of their right and left biceps femoris, right and left longissimus dorsi, myocardium, and tongue were frozen at -150 C in precooled isopentane and stored at -70 C. Frozen sections of these muscles were stained with hematoxylin and eosin, modified Gomori's trichrome, nonspecific esterase, diphosphopyridine nucleotide tetrazolium reductase, and adenosinetriphosphatase at pH 10.4 and 4.6. Muscle from the same locations was fixed in 10% neutral buffered formalin, processed for paraffin embedding, sectioned, and stained with hematoxylin and eosin. Microscopic examination of both calf and goat tissue indicated that both type I and type II muscle fibers were equally infected and that infected myofibers showed no apparent damage other than displacement by sarcocysts. Occasionally, muscle fibers within the muscle spindles contained sarcocysts.     

A Study of the Number and Distribution of Cutaneous Mast Cells in Cats with Disease not Affecting the Skin

Resumen— El propósito de este estudio era investigar el número de mastocitos dérmicos en diferentes partes de la piel de gatos con enfermedad que no afecta a la piel. Se hizo un recuento de el número de mastocitos en 10 partes diferentes de la piel de 12 gatos sin evidencia de enfermedad de la piel. Se fijaron muestras de biopsia de piel en fijador Carnoy y formalina neutral atenuada, y se tiñeron con azul touluidina. Se realizaron recuentos de celulas de 6 campos de alto poder (400), de cada parte. Los recuentos de mastocitos combinados para todos los gatos, todas las partes y ambos fijadores variaban de 0–60 con un promedio de 12.5 por campo de alto poder y eran mas altos en el aspecto caudal del pabellón auricular y en la barbilla. El tipo de fijador usado no pareció influir en el recuento de mastocitos. [Foster, A. P. A study of the number and distribution of cutaneous mast cells in cats with diseases not affecting the skin (Un estudio de el número y distribución de mastocitos cutaneos en gatos con enfermedad que no afecta a la piel).
Abstract— The purpose of this study was to investigate the number of dermal mast cells in different skin sites from cats with disease not affecting the skin. The number of mast cells in the skin of 12 cats without evidence of skin disease was counted from 10 different skin sites. Skin biopsy samples were fixed in Carnoy's fixative and neutral buffered formalin and stained with toluidine blue. Cell counts from 6 high power fields (400) were made for each site. Mast cell counts combined for all cats, all sites and both fixatives ranged from 0 to 60 with a mean of 12.5 per high power field and were highest from the caudal aspect of the pinna and the chin. The type of fixative used did not appear to influence mast cell counts.     

Changes in serum thyroid hormone levels in newborn calves as a diagnostic index of endemic goiter

Maximum serum thyroxine (T4) and triiodothyronine (T3) levels of healthy calves were seen at 1 day after birth, and thereafter rapidly decreased until 5 days after birth. They stabilized until 2 weeks after birth, then gradually decreased until 4 weeks after birth. Serum T4 levels of calves with endemic goiter tended to be lower than those of healthy ones, but showed similar levels to those of adult cows. T3 levels of calves with goiter were similar to those of healthy ones, but showed higher variation. T4/T3 ratio of calves with goiter were significantly lower than those of healthy ones and adult cows. While individual levels of serum T4 and T3 at just after birth could not be considered as a diagnostic index, the T4/T3 ratio could be adopted as a diagnostic index of endemic goiter.     

A modified immunoperoxidase assay for detection of antibody porcine reproductive and respiratory syndrome (PRRS) virus in swine sera

MARC-145 cell monolayers infected with PRRS virus were fixed in 3% neutral buffered formalin and embedded in paraffin. The sections were stained by avidin-biotin complex immunoperoxidase (ABC) method. Test sera were applied to sections as primary antibodies. The positive reactions were detected by ABC method and indirect immunofluorescent assay (IFA). There was good correlation between ABC and IFA, and the titers in ABC were higher than those in IFA. The present results indicate that the immunohistochemical staining is a useful test for the detection and quantitation of PRRS virus antibody in swine sera as well as IFA.     

Sirius red is able to selectively stain eosinophil granulocytes in bovine, ovine and equine cervical tissue

The aim of the present study was to examine the suitability of sirius red staining for selective light microscopic demonstration of eosinophil granulocytes in cervical tissue of mares, cows and sheep. For this purpose, tissue was fixed in 4% neutral buffered formol or in Bouin's solution. Paraffin sections of 5-microm thickness were stained with sirius red. In cows, mares and sheep a selective distinction of eosinophilic infiltration is successful after both fixation methods.     

Response of calves to exposure with swine influenza virus

In calves inoculated with live swine influenza virus (SIV) A/sw/IL/75 (H1N1) intranasally, SIV was isolated for 7 days, and respiratory tract disease was observed. Antibody was detected in serum of inoculated calves from postinoculation day 9, and virus-neutralization antibody was demonstrated on postinoculation days 14 and 21. The primary response was low, but readily differentiated from the secondary response after calves were challenge exposed with homologous SIV. Pneumonic lesions were observed at necropsy, and histopathologic changes in airways and lungs were consistently found. Fluorescent staining revealed viral activity in epithelial cells of airways. The virus was transferred to healthy calves housed with inoculated calves.