Establishment of a cell line (MCM-B2) from a benign mixed tumour of canine mammary gland

A cell line was established from a benign mixed tumour of the canine mammary gland. Light microscopy of the cells cultured on plastic dishes revealed monolayer colonies. Cells that grew within the collagen gel matrix formed large three-dimensional colonies with a branching pattern. Immunohistochemically, these cells reacted intensely with anti-vimentin antiserum and mildly with anti-desmin antiserum. Ultrastructural examination revealed a large nucleus, intracytoplasmic organelles and intermediate filaments, which varied among cells. The cells possessed an abnormal chromosome number, an average of 80 per cell. Histologically, the xenografted tumour of cultured cells was similar to anaplastic carcinoma and reacted strongly with antivimentin antiserum, mildly with anti-desmin antiserum, and weakly with anti-keratin antiserum. The average chromosome number of cells form the xenografted tumour was the same as that of the original cultured cells. These findings suggest that the cell line might be derived from stem cells or atypical cells, and that it should be useful as a model for the study of cell differentiation and proliferation in canine mammary tumours.     

Leiomyosarcoma of the kidney in a dog

A leiomyosarcoma that occurred in the right kidney of a dog (7-year-old female Labrador retriever) was studied pathologically. On histological examination, tumour cells were spindle-shaped, the cell density was high, and complex fasciculated tumour cells showing longitudinal and transverse cross-sections were observed. Both ends of the nuclei were rounded, the nucleoli were clear and many images of nuclear division were observed. Immunohistochemically, tumour cells reacted intensely with anti-vimentin antibody and anti-actin smooth muscle antibody, and mildly with anti-desmin antibody. On electron microscopy, the nuclei of tumour cells were rounded at both ends and elongated elliptically, and a large number of microfibrils with focal density and dense patches were observed in the cytoplasm. This is a case report of a very rare leiomyosarcoma of the dog kidney.     

A malar granular cell tumor in a djungarian hamster

'Granular cell' tumor observed in the malar subcutis of a Djungarian hamster was examined to determine its cellular origin. Histologically, the tumor consisted of a solid growth of oval or spindle-shaped large cells with abundant cytoplasm filled with eosinophilic granules that were periodic acid-Schiff-positive and diastase-resistant. Immunohistochemically, the tumor cells were positive to anti-vimentin and anti-desmin antibodies and a few cells showed positivity to anti-actin antibody as well. They did not react to myoglobin, S-100 protein, and glial fibrillary acidic protein (GFAP). Electron microscopic studies revealed that the tumor cells had pinocytotic vesicles, dense plaque and microfilaments. The first granular cell variant of myogenic tumor reported here in Djungarian hamsters was differentiated from granular cell tumor of Schwann cell origin.     

实验性鸡马立克氏病羽髓病变研究

通过光镜1电镜等方法对鸡马立克氏病羽髓组织进行了形态学研究。结果显示，羽髓远端坏死，羽髓中有多量肿瘤细胞浸润，肿瘤细胞呈弥漫性或局部性浸润，电镜观察，肿瘤细胞核异型性明显，核膜凹陷，核内出现假核内包含物，在一些羽髓中，巨噬细胞吞噬肿瘤细胞较多见，而且观察到一些肿瘤细胞发生调亡，结果表明，鸡马立克氏病羽髓病变十分明显，羽髓的形态学变化可用于鸡马立克氏病的诊断。     

Establishment and characterization of eight feline mammary adenocarcinoma cell lines

Eight new feline mammary adenocarcinoma cell lines derived from either primary or metastatic lesions were established. The morphology of all the cell lines was epithelioid and round to spindle in shape, with cell growth occurring in a monolayer fashion. On immunohistochemistry, these cells reacted with anti-keratin and anti-vimentin antisera. The doubling time of these cells was between 19 and 54 hr. Tumor masses were developed in nude mice by subcutaneous inoculation of the cells that were histologically identical to their original mammary tumor lesions. Telomerase activities measured using the telomeric repeat amplification protocol assay revealed high telemetric activity in all of the cells.     

Psittacine beak and feather disease in three captive sulphur-crested cockatoos (Cacatua galerita) in Thailand

Three sulphur-crested cockatoos (Cacatua galerita) were diagnosed as psittacine beak and feather disease (PBFD). Histopathology of the feather pulp and follicles showed intracytoplasmic botryoid clusters or granular inclusion bodies in epithelial cells and macrophages. Electron microscopy revealed multiple cytoplasmic clusters of electron dense viral particles corresponding to the inclusions. PBFD virus (circovirus) DNA-specific product was detected from formalin-fixed paraffin-embedded feathers by nested polymerase chain reaction (PCR) method.     

The pathology of psittacine beak and feather disease

Psittacine beak and feather disease is characterised by loss of feathers, abnormally shaped feathers and overgrowth and irregularity of the surface of the beak. The disease occurs in a number of psittacine species including the Sulphur-crested Cockatoo, Lovebirds , Budgerigars and Galahs . The abnormal appearance of feathers and beak is due to a dystrophic process within the epidermis of the feather and beak. The process consists of epidermal cell necrosis, epidermal hyperplasia and hyperkeratosis. Many of the feather abnormalities are due to retention of a hyperkeratotic feather sheath. A characteristic microscopic finding is the presence of macrophages containing purple intracytoplasmic inclusions in affected epidermis and feather pulp. The inclusions consist of aggregates of particles 17 to 22 nm in diameter. Similar but smaller inclusions occur in epidermal cells. In addition, non-suppurative inflammation occurs in the feather pulp. The findings are suggestive of a viral infection.     

A Simple and Rapid PCR‐Based Method for Ostrich Sexing Using Micro Amounts of DNA

The aim of the present study is to identify ostrich sex by using polymerase chain reaction (PCR) on micro amounts of DNA from blood, bloodstain and feathers. Sixteen male and 18 female ostriches were used as test individuals. Genomic DNA as a template was extracted by the Chelex method. Ostrsex‐P1 and P2 primers were designed to perform PCR amplification on the template. PCR products were checked using agarose gel electrophoresis with ethidium bromide staining and ostrich sex was determined directly by the bands shown on the gel. The results demonstrate that ostrich sex can be determined by the extraction of DNA from as little as 0.0125 μl blood using Chelex, whereby the use of large amounts of organic solvents such as phenol and chloroform are unnecessary. In addition, it is possible to identify ostrich sex using micro amounts of DNA extracted from bloodstains and/or feathers. The use of feathers particularly avoids unwanted sampling problems such as the difficulty of collecting ostrich blood, the stress to the ostrich caused by bleeding, and the demand for a lot of manpower for ostrich restraint.     

The identification of intestinal M cells in the sacculus rotundus and appendix of the Angora rabbit

The present study was aimed at the immunohistochemical demonstration of M cells, found in the follicle-associated epithelium (FAE) of the sacculus rotundus (SR) and appendix of the Angora rabbit, using anti-vimentin primary antibodies, and at the determination of certain fine structural characteristics. Ten adult Angora rabbits constituted the material of the study. Immunohistochemical staining revealed that many cells composing the FAE, which covered the dome regions of the SR and appendix, reacted positively with vimentin. FAE contained two different types of vimentin-positive cells. The first type surrounded intraepithelial lymphocytes (IEL) with a basolateral invagination in the apex and periphery of the dome epithelium, whilst the second type consisted of columnar cells found in the FAE near crypts. The immunoreactivity of the cells found in the FAE covering the apex and periphery of the domes was observed particularly in the perinuclear cytoplasm and the cytoplasm surrounding the IEL. Electron microscopic examination demonstrated that the M cells found in the FAE covering the apex and periphery of the dome regions of the SR and appendix did not exhibit any microvilli on their apical surface. The FAE near crypts contained columnar cells, which resembled enterocytes. The apical membrane of these cells exhibited shorter and irregular microvilli, in contrast to neighbouring enterocytes. It was determined that M cells, found in the FAE of the SR and appendix in the Angora rabbit, displayed similarities in terms of localization and fine structure. This situation may be indicative of the two lymphoid structures with different localization having similar functional properties.     

A description of the growth and moulting of feathers in commercial broilers

ABSTRACT

1. Changes in feather length and weight and daily losses of down, contour feathers, remiges and retrices were studied in two commercial broiler strains to determine feather growth and moult in broilers up to 112 d of age.

2. Ten pens of 20 chicks for each sex x strain were fed adequate amounts of dietary protein in a four-phase feeding programme. Ten birds were sampled per genotype at 14, 28, 42, 56, 70, 84, 98 and 112 d. Feather loss was determined for individual birds caged within a nylon net.

3. All feathers were dry-plucked from each of seven tracts, with representative feathers from the capital-cervical, dorsopelvic and interscapular, pectoral and femoral tracts being randomly selected. Three rectrices of the dorsocaudal tract and three primaries and secondaries of the humeral-alar tract of the right wing were evaluated.

4. The length of the selected feathers was measured, and the feathers from each tract and from the whole bird were weighed.

5. A wide range of rates of maturing (0.0250–0.0907/d) and mature weights (9.62–52.9 g) were evident between sampled tracts.

6. Feather weight failed to predict some moults which were detected by the measurement of feather losses. From the weight data, moulting was evident only in the humeral-alar and dorsocaudal tracts, whereas, when daily losses were measured, contour feathers accounted for over 0.7 proportion of feather losses. Logistic equations adequately described the cumulative losses of down, contour feathers and remiges.

7. Feather loss needs to be considered when feather growth is determined from feather weight at different ages.

8. The rate of maturing (B) of feathers was numerically higher in Ross males and females than in the Cobb strain. In females, B was higher than in males (0.0483 vs. 0335/d) but the mature weight was lower (162 vs. 230 g).     

Utilization of citrus pulp based diets and Enterolobium cyclocarpum (JACQ. GRISEB) foliage by West African dwarf goats

Sixteen West African dwarf goats weighing 5.0–5.6 kg were used in an experiment that lasted 112 days to investigate the utilization of citrus pulp based diets and E. cyclocarpum. A control diet which contained 88.5% dried brewers' grains (DBG) and other three experimental diets in which DBG was replaced with dried citrus pulp (DCP) at 25, 50 and 75% levels, respectively constituted the four treatments. The goats were divided into four groups of four animals each to balance for variation in live weight before they were assigned to the four different experimental diets. Significant differences (P < 0.05) were obtained in growth rate with (22.14–34.02 g/d) the highest and lowest value from 50 and 75% DCP inclusion respectively. The digestibility coefficients of DM, Ash, NFE, NDF, ADL, cellulose and hemicellulose increased with increasing levels of dried citrus pulp in the diets and were highest at 75% level of replacement. However, the CP digestibility of 83.85% was highest in the control diet and was significantly (P < 0.05) different from other diets containing citrus pulp. Nitrogen intake (g/d) decreased significantly (P < 0.05) with increasing levels of dried citrus pulp in the diets from 45.57–33.28 and was lowest in the control diet. Nitrogen balance also followed the same trend, being highest (42.07 g/d) at 25% DCP and lowest at 75% DCP. The Packed cell volume (PCV), haemoglobin concentration (Hb), red blood cells (RBC), white blood cells (WBC) and total blood protein (TBP) were significantly (P < 0.05) different across the dietary treatments at the start and end of the trial. The values for PCV, RBC, MCH, glucose and total protein decreased slightly in the control diet. Serum glutamate pyruvate transaminase (SGPT) values also increased significantly (P < 0.05) with increasing levels of dried citrus pulp in the diets at the end of the experiment and were highest (6.50 IU/L) at 75% level of citrus pulp. Therefore, the efficient utilization of citrus pulp and E. cyclocarpum by WAD goats was attained at the 50% level of inclusion in the diets.     

Metal accumulation in House Sparrow (Passer domesticus) from Thohoyandou,Limpopo province,South Africa

Studies on metal pollution (cadmium, cobalt, chromium, copper, manganese and nickel) in South African terrestrial environments are severely lacking. Due to being relatively unaffected by industrialisation, the Thohoyandou region may provide data on natural levels of metals for use as baseline data. The House Sparrow (Passer domesticus) was chosen as a bio-indicator of metal pollution due to its abundance, non-migratory lifestyle and close association with humans. The aims were to determine the viability of using feathers as a non-lethal bio-indicator tissue compared with muscle. Plume feathers, flight feathers and muscle tissue were analysed using ICP-OES techniques. Analyses of tissue metal concentrations identified the following trend: plume feather > flight feather > muscle tissue. Within the Thohoyandou region, Magondi, which was affected by anthropogenic activities at the time of sampling, had significantly higher concentrations (p ≤ 0.05) for each of the metals, indicating potential metal contamination from various sources. Alternatively, Makonde had the lowest metal concentrations and may be an important reference site for future comparative studies. The results showed that the House Sparrow can be used as a bio-indicator organism in South African terrestrial environments. The plume feathers indicate that they are a good non-lethal tissue for determining metal pollution levels on a temporal scale, indicating recent metal exposure.     

Histological and Immunohistochemical Study of the Cat Epididymis

A histological and immunohistochemical study of the epididymis of 21 cats which ranged from 7 to 8 months to 2 years of age was made. Five different cell types were observed in the feline epididymis: principal and basal cells, which were the most numerous, apical and narrow, PAS positive cells, which were scarce, and migratory cells, consisting of lymphocytes and macrophages. Four morphologically distinct areas of the cat epididymis were identified. Region I displayed a stellate lumen and the principal cells were tall and presented long and irregular stereocilia. In region II, the lumen was oval and the principal cells were shorter than in the inicial area. Region III displayed characteristics similar to those of region I; the principal difference was the presence of short and regular stereocilia on the surface of the principal cells. In region IV the lumen was very spacious and the epithelium shorter than in the other regions. In regions I, II, and III intraepithelial cavities were observed. With regard to the immunohistochemical results, the basal cells displayed medium immunoreactivity with vimentin. Due to its anti-desmin reactivity, the muscle wall which surrounds the epididymis was seen to become progresively thicker as it nears the tail. Broad-spectrum anti-cytokeratin serum produced intense immunostaining in the basal, apical and narrow, PAS positive cells; the principal cells of region IV also displayed a strong immunoreactivity.     

Nutrient digestibility and evaluation of protein and carbohydrate fractionation of citrus by‐products

The protein and carbohydrate fractionation and nutrient digestibility of citrus by‐products were determined. Ruminal, intestinal and total tract CP disappearance values were measured by a modified three‐step (MTSP) method and in vitro CP disappearance method (IVCP). Test feeds were orange pulp (OP), lime pulp (LP), lemon pulp (LEP), grapefruit pulp (GP), sweet lemon pulp (SLP), bitter lemon pulp (BLP), bergamot orange pulp (BP) and tangerine pulp (TP). The rumen undegradable protein (RUP) fractions of the feedstuffs were obtained by ruminal incubation in three cannulated wethers and incubation in protease solution (protease type xiv, Streptomyces griseus). The data were analysed using completely randomized design. There were significant differences between the tested feeds in protein fractions and acid detergent insoluble nitrogen (ADIN; C fraction) was highest in GP (14.56%) (p < 0.001). For carbohydrate fraction, the highest C fraction was also observed in GP (2.67%) and in relation to the other citrus pulps (p < 0.001). Ruminal CP disappearance was highest in OP (71.89%) (p < 0.001). The level of post‐ruminal CP disappearance, measured by MTSP, was highest for BP (34.94%) (p < 0.001). The highest in vitro dry matter digestibility (IVDMD) was found for TP (80.44%) followed by that estimated for BP (78.38%) (p < 0.001). The estimated metabolizable energy (MJ/kg DM) varied from 9.77 for LP to 12.91 for BP. Tangerine pulp had the highest true rumen digestibility (TRD) (p < 0.001). According to the results, it could be concluded that citrus by‐products have high nutritive value and also, the in vitro techniques can be easily used to determine of the nutritive value of citrus by‐products.     

A novel DNA virus associated with feather inclusions in psittacine beak and feather disease

The nature of feather inclusions was characterized in 32 psittacine birds (30 cockatoos, one peach-faced lovebird (Agapornis roseicollis), and one red-lored Amazon parrot (Amazona autumnalis autumnalis] with naturally-acquired psittacine beak and feather disease. Intranuclear inclusions within feather epithelial cells and intracytoplasmic inclusions within macrophages in the feather epithelium and pulp cavity contained psittacine beak and feather disease viral antigen when stained by the avidin-biotin complex immunoperoxidase technique. Ultrastructurally, inclusions were observed primarily within macrophages and to a lesser extent within epithelial cell nuclei. Macrophage inclusions appeared as paracrystalline arrays of viral particles. Intranuclear inclusions were less well defined, although scattered viral particles were present. Intracytoplasmic and intranuclear particles in ultrastructural preparations were identified by colloidal gold labeling as psittacine beak and feather disease virus. Feather epithelium was more frequently and severely involved in the disease process than was adjacent follicular epithelium. Plucked feathers with an intact epidermal collar and feather epithelium were preferred to follicular biopsies for histopathologic examination.     

Fine Structural and Histochemical Study of Equine Paneth Cells

Ultrastructure. lysozyme and glycoconjugate activity in duodenal Paneth cells were observed concurrently in the horse. Paneth cells were seen to uniformly line the base of the equine intestinal glands. The round secretory granules have centrally located electron densities with peripherally located electron lucent halos. Histochemically, the peripheral halo layer was positively stained for carbohydrates by the periodic acid-thiocarbohydrazide-silver protein-physical development (PA-TCH-SP-PD) method and the entire granules reacted positively to the WGA. The central core area reacted with anti-lysozyme. We identified a young (Type I) and an old (Type II) cell population in the same crypt, but we suggest that the observed populations are variations of the same cell type with the varied appearance due to aging of the secretory granules.     

Depletion study of three formulations of flumequine in edible tissues and drug transfer into chicken feathers

Cornejo, J., Lapierre, L., Iragüen, D., Pizarro, N., Hidalgo, H., San Martín, B. Depletion study of three formulations of flumequine in edible tissues and drug transfer into chicken feathers. J. vet. Pharmacol. Therap. 34 , 168–175. To ensure the delivery of safe animal products to consumers, withdrawal times (WDT) of drugs must be respected. Drugs administered in therapies can also reach nonedible tissues (for humans) such as feathers; this transfer is of concern as feather meal is used in diets of food producing animals, being this a possible source of residue contamination of final products for human consumption. WDTs of three flumequine formulations (10%, 80% premix powder and 20% solution) as well as the transfer of this drug into feathers were determined. One hundred and twenty broiler chickens were allocated into four experimental groups (36 birds each). Three of them were treated with 24 mg/kg bw orally for five consecutive days of each flumequine formulation, whereas one group remained untreated (12 birds as control group). After the treatment ended, six chickens of each experimental group and two controls were slaughtered daily for 6 days. Samples of muscle, liver and feathers were collected and analyzed by liquid chromatography tandem mass spectrometry (LC MS/MS). The WDTs showed differences between formulations. Flumequine concentrations found in feathers remained high during WDT and after this period, thus suggesting that the WDTs estimated for the pharmaceutical formulation of flumequine do not guarantee the absence of this drug in chicken nonedible tissues such as feathers.