Paraffin Embedding Technique Research Articles

Histopathological Profile of Cholecystectomy Specimens at a Tertiary Care Center: A Retrospective Study

Abstract Introduction: Gallbladder is one of the most frequently received specimens in any histopathology laboratory. Its diseases may present with a varied spectrum ranging from congenital anomalies, cholelithiasis, and inflammatory or noninflammatory lesions to noninvasive and invasive neoplasms. Cholelithiasis (gallstones) is the most common lesion and accounts for more than 95% of gallbladder diseases and affects 10%–20% of the adult population. The incidence is 2–4 times higher in females than in males. Aims and Objectives: This study was designed to study the histopathological spectrum of gall bladder diseases in the population, study the most common gall bladder diseases in cholecystectomy specimens, and study the frequency of gall bladder carcinoma. Methodology: This retrospective observational study was done in the Pathology department at a tertiary care hospital and research center, from September 2016 to September 2023. Tissue was fixed in 10% buffered formalin and then processed by the routine paraffin embedding techniques. Sections were cut at 4–5 μ thickness and stained with hematoxylin and eosin stains. Special stains were done if required for confirmation. All the slides were analyzed and reported by a trained histopathologist. Results: Females were affected more as compared to males. The commonest age group affected was 41-50 years. Gallstones were observed in 33.58% of patients. Gallbladder neoplasm was rare and incidental findings. Conclusion: Gallbladder disease demonstrated a diverse spectrum of histopathological changes in the cholecystectomy specimens and showed female preponderance. Gallbladder carcinoma was observed as a rare occurrence and was an incidental finding mainly. Hence, a microscopic (histopathological) examination is required for every cholecystectomy specimen.

Anatomía del pecíolo de 16 especies de Polypodiaceae presentes en México

Background and aims: Several works have shown that the petiole possesses useful anatomical characteristics for differentiating groups. Therefore, we carried out a descriptive and comparative anatomical study of the basal, middle, and apical regions of the petiole of 16 species belonging to 8 genera of Polypodioideae growing in Mexico, to identify anatomical characters that can contribute to their delimitation and identification M&M: Samples were collected from the basal, middle, and apical regions of the petiole and processed according to the paraffin embedding technique to obtain permanent preparations. The generated information was evaluated through a clustering analysis and a principal component analysis. Results: All petioles showed an unistratified epidermis, a cortex differentiated into external and internal, and vascular bundles with Polypodium-type distribution. Differences were registered in the petiole diameter, indument type, circumendodermal band, and number of vascular bundles. Four types of external cortex are recognized and the presence of nectaries in Campyloneurum amphostenon and C. costatum is reported for the first time, as well as the glandular hairs inserted at the base of the scales in Microgramma percussa. Identification keys were elaborated. Conclusions: The anatomical characteristics of the petiole allow us to differentiate taxonomically the genera and most of the species analyzed, the most important traits being the petiole diameter, the characteristics of the circumendodermal band, and the presence of aerophores and/or wings

Caecal villi? A comparative histological and morphometric study of caecal and jejunal mucosa in adult rabbits

BackgroundRabbits are herbivores with a distinctive digestive strategy that differs significantly from other caecal fermenters (e.g., horses, guinea pigs) and ruminants. In view of this, the current study aimed to highlight distinctive histological and morphometric features of the caecal mucosa in adult rabbits that accentuate its major role in digestion. The caecal and jejunal samples were harvested from five 1-year-old domestic rabbits and processed by regular paraffin-embedding histological technique followed by Goldner’s trichrome staining. A comprehensive morphological and morphometrical analysis of the jejunal mucosa vs. caecal mucosa was performed.ResultsMicroscopically, as in the case of the jejunal mucosa, the caecal mucosa presents long and often branched finger-like villi covered by a simple columnar epithelium mostly made of enterocytes with a prominent microvillous brush border. Besides, the caecal villi include a lacteal along with the villous muscle. Statistically, except for villus length, all the parameters assessed in the caecal mucosa, including villus width, villus count, thickness of the brush border and enterocyte/goblet cells ratio, revealed a high grade of similarities with the jejunal villi.ConclusionsAccording to the obtained results, the caecal mucosa in adult domestic rabbits includes unique features, namely caecal villi, structures infrequently presented in the large intestine of other adult mammals. Those structures once more emphasize the major role of the caecum not only in fermentation but also subliminally in local absorption. To our knowledge, this is the first reliable microanatomical and morphometric report of caecal villi in adult domestic rabbits.

Influence of sexual hormones on Chagas disease.

Analyze sex hormone's influence during Chagas´ Disease. Male and female BALB/c mice were divided into six groups, four experimental (sham, orchiectomized, orchiectomized and supplemented with estradiol, orchiectomized supplemented with testosterone, oophorectomized, oophorectomized and supplemented with estradiol, and oophorectomized and supplemented with testosterone), and two control (healthy and intraperitoneally with T. cruzi strain NINOA infected). Clinical data were recorded daily, parasitemia was evaluated using a Neubauer chamber during the infection, and heart histopathological analysis was performed using the paraffin embedding technique. To analyze parasitemia curves and the area under the parametric curves, two-way ANOVA test was performed to correlate groups´ data. P-values <0.05 were considered statistically significant. Higher mortality rates, cardiomegaly, hepatomegaly, ascites, edema, higher parasitemia levels, more amastigote nests, and more severe inflammatory infiltrate were found in higher testosterone concentration mice, whereas in higher estradiol concentration groups, paresia, prostration, edema, and necrosis were found. Our results showed that testosterone increased infection severity, whereas estradiol had the opposite effect. This research improves the understanding of sex hormones´infuence upon this infection to contribute with the handling of Chagas´disease.

Conventional paraffin embedding technique and double-embedding technique for minute oral biopsies and delicate pulp tissue - a comparative study

Among oral biopsies, small incisional tissues, have to be preserved all through the processing and embedding to ensure optimal visualization of all the mucosal layers without compromise. Optimal tissue orientation is the most critical step in tissue processing for demonstration of definitive morphology in the sections, which is often more challenging in cases of minute/small or thinner sections using routine paraffin techniques to evaluate accurate diagnosis. Some modification is needed to handle these samples to get a better result. Double embedding technique with some modification has been widely used for small/ thin/ multiple biopsies and gives excellent results in many other fields like general pathology and biotechnology. The double embedding technique though produced excellent and significant results in mucosal biopsies yet, it is of minimal interest among oral pathologists. To best of our knowledge, this is the first study to use double embedding technique for pulp tissues. The present study was aimed to evaluate and compare the ease of embedding and sectioning sections using Agar-Paraffin double embedding technique for small oral mucosal biopsies and thin pulp tissues. A total of 40 oral tissue samples categorized into two groups were taken for the present study. Group I included 20 small oral mucosal biopsy samples of size ranging from 0.2 to 0.5 cm and Group II included 20 pulp tissues obtained from freshly extracted non carious tooth. 10 blocks were prepared by routine paraffin method and 10 blocks were prepared by modified double embedding method for each group. Scores were given by comparing all the criteria with that of the routine paraffin technique. Chi-square test was used for statistical analysis. The average ease score for the Agar-Paraffin double embedded small/minute biopsies showed better scores than the pulp tissue with that of the routine technique. However, no statistically significant difference was seen among embedding and sectioning sections between the two groups. Modified double embedding method is simple and reliable alternative technique that helps in better orientation, processing and sectioning especially for oral small or thin biopsies and delicate pulp tissues.

Seasonal changes in the scrotal skin histology in relation to thermoregulation and testosterone level in camel and buffalo bull.

This study was conducted to compare the scrotal skin dynamics of camel and buffalo bulls regarding thermoregulation, season and testosterone. Scrotal skin and blood samples (n = 8 each) were collected during breeding (October to March) and non-breeding seasons (April to August). Tissue slides were prepared by the paraffin embedding technique followed by hematoxylin and eosin (H&E) staining and were analysed by ImageJ®. Relative humidity and temperature were recorded in different seasons to calculate the thermal humidity index (THI). Serum testosterone level was estimated through radioimmunoassay (RIA). The data were analysed by applying ANOVA, the THS test was used as a post-ANOVA interface and the correlation coefficient was also calculated among season, testosterone and skin parameters. The results showed that skin thickness, season and THI were negatively correlated with testosterone levels. In the neck, body and apex, the papillary layer was thicker in buffalo bull compared to camel bull during the breeding season; however, the reticular layer followed a reverse trend in the apex when THI was low. In the scrotal neck during the non-breeding season, hair follicle (HF) length and density were recorded high in camel bull compared to buffalo bull when THI was elevated. The density of HF followed the otherwise trend in the apex during the breeding season. The density of SG was found highest in all regions of camel bull as compared to buffalo bull and showed direct relation with THI. To conclude, this study delineates how the histo-dynamics of scrotal skin varies and accommodate itself with testosterone and THI over the different season in camel and buffalo.

Potential application of improved hard tissue section technique in the clinical pathological diagnosis of bone and bone marrow

Objective: To discuss the application value of hard tissue section in the clinicopathology diagnosis. Methods: From March 2021 to December 2021, bone slices of 19 patients (1 patient with osteochondroma, 2 patients with chondrosarcoma, 4 patients with osteosarcoma, 2 patients with fibrous dysplasia, 2 patients with bone metastasis from thyroid papillary carcinoma, 2 patients with osteomyelitis, 4 patients with giant cell tumor of bone, 2 patients with Ewing sarcoma) and 16 hemopathy patients were collected from the Department of Pathology, Shanghai Sixth People's Hospital. Of the osteopathy patients, there were 14 male and 5 female with a median age of 31 (10-66) years. Meanwhile, there were 7 male and 9 female with a median age of 28 (16-65) years among these hemopathy patients. Thirty-five cases were treated with modified hard tissue slicing technique and paraffin embedding technique, respectively. The advantages and disadvantages of the two methods for clinical diagnosis of bone disease were compared by Hematoxylin-Eosin staining (H&E staining), immunohistochemical staining (IHC), fluorescence in situ hybridization (FISH) and Sanger sequencing. Results: The improved resin-embedded method showed better histological morphology and cell structure. Besides, the expression of Ki67, SATB2, CD34, SMA, CD68,MPO,CD4 and CD33 in immunohistochemical staining in bone tissues which were embedded in resin were more clear in the accurate positive localization than those using paraffin-embedded. MDM2 of FISH exhibited a higher fluorescence intensity and more accurate location. Meanwhile, both methods treated with Sanger sequencing met the requirements of DNA purity and mutation detection. Conclusion: The improved hard tissue section method is simple and short time-consuming, which is suitable for optimizing the clinical bone and bone marrow pathological diagnosis process.

Histometric and morphological damage caused by Serratia marcescens to the tick Rhipicephalus microplus (Acari: Ixodidae).

Rhipicephalus microplus tick is the ectoparasite causing the greatest economic losses in the livestock industry. Multi-resistance in ticks is increasing, generating the inefficiency of traditional ixodicides, for which biological control has been proposed as an alternative. In this work, we analyze the histomorphological damage caused by the bacterial strain EC-35 on Rhipicephalus microplus. The ixodicidal effect of EC-35 total protein was evaluated on larval or adult ticks comparing with the commercial ixodicide coumaphos 0.02% as a control. Female ticks were processed using the paraffin-embedding technique and stained with hematoxylin-eosin. Also, the pathogenicity of EC-35 was evaluated by capillary feeding and coelom inoculation tests. The identification of the bacterium was performed using the molecular markers 16S RNA and rpoB, by PCR and sequencing technique, and the evolutionary distance was analyzed by Bayesian phylogenetic inference. No differences were observed in the perimeter and area of larvae treated with EC-35 or Coumaphos. The thickness of the integument decreased a 65% with the EC-35 treatment (6.01 ± 0.6µm) and of 30% in coumaphos (12.04 ± 1.2µm) in larvae compared with the control group (18.41 ± 2µm), while no difference was found in adult ticks. The capillary feeding test and coelom inoculation with EC-35 showed an inhibition of reproductive potential of 99.8 ± 7 and an oviposition Inhibition 97 ± 3.02%. The EC-35 strain was genetically related to Serratia marcescens, concluding that these bacteria caused high mortality, oviposition Inhibition, and integument thinning and drastic loss of histoarchitecture in R. microplus tick larvae.

The histological and histochemical features of the esophagus in local breed dogs (Canis familiaris)

A total of eight samples of the esophagus of local breed dogs are used to investigate the histochemical features of the esophagus. The specimens are processed according to paraffin embedding technique protocols and the tissue sections are stained with hematoxylin and eosin, and masons trichrome combined with Alcian blue (pH2.5)-PAS. The esophagus folds the mucosa which is covered by keratinized to non-keratinized stratified squamous epithelium which contains numerous excretory common ducts of esophageal glands. The epithelial thickness of cervical, thoracic and abdominal parts of the esophagus are 221.95±3.41µm, 212.46±5.38 µm, and 173.15±4.09 µm respectively. The lamina propria of the cervical part of the esophagus lacks the muscularis mucosa, while the muscular mucosa in the thoracic part of the smooth muscles appears as scattered interrupted bundles. The esophageal glands are a type of compound tubular mucoserous constructed of the predominated mucous alveoli and little serous acini. When combined with Alcian blue (pH2.5)-PAS stain, the esophageal glands show strong acidic mucopolysaccharides within the mucous alveoli and light blue color within serous acini that denote weak acidic zymogen granules. Tunica muscularis is striated muscle fibers in cervical and thoracic parts and smooth in the abdominal part of the esophagus. The measured thickness of tunica muscularis of cervical, thoracic and abdominal parts are 568.76±6.90 µm, 703.29±7.54µm, and 338.98±7.26 µm respectively.

Histomorphological changes in the goat, &lt;em&gt;Capra hircus &lt;/em&gt;L. rumen during different stages of gestation

Understanding the fetal development is essential to treat different anomalies of livestock after birth. Biological and anatomical data on the developing rumen of goat is scanty. Here, we investigated histomorphological changes in the ruminal part of the fore stomach of the goat, Capra hircus L. to elucidate the transformation of rumen prior to its functional state. For this study, 36 developing rumen were collected from healthy and normal embryos/foeti of either sex of goat from the first stage of prenatal life until birth. Tissues were processed by paraffin embedding technique and sections were stained. Definite four layers of the tubular structure were elucidated first at 51 days of gestation. We observed the ruminal wall lined by undifferentiated stratified epithelium up to 100 days which gradually became stratified squamous keratinized epithelium at 145 days of foetal life. Throughout the gestation lamina muscularis was absent. Reticular fibers evident first followed by collagen and elastic fibers. The prenatal development of the goat ruminal mucosa evidenced a considerable resemblance with the post natal rumen; however, certain morphological changes are still incomplete in order to meet the functional demands in postnatal life

Maintaining Human Glioblastoma Cellular Diversity Ex vivo using Three-Dimensional Organoid Culture.

Glioblastoma (GBM) is the most commonly occurring primary malignant brain cancer with an extremely poor prognosis. Intra-tumoral cellular and molecular diversity, as well as complex interactions between tumor microenvironments, can make finding effective treatments a challenge. Traditional adherent or sphere culture methods can mask such complexities, whereas three-dimensional organoid culture can recapitulate regional microenvironmental gradients. Organoids are a method of three-dimensional GBM culture that better mimics patient tumor architecture, contains phenotypically diverse cell populations, and can be used for medium-throughput experiments. Although three-dimensional organoid culture is more laborious and time-consuming compared to traditional culture, it offers unique benefits and can serve to bridge the gap between current in vitro and in vivo systems. Organoids have established themselves as invaluable tools in the arsenal of cancer biologists to better understand tumor behavior and mechanisms of resistance, and their applications only continue to grow. Here, details are provided about methods for generating and maintaining GBM organoids. Instructions of how to perform organoid sample embedding and sectioning using both frozen and paraffin-embedding techniques, as well as recommendations for immunohistochemistry and immunofluorescence protocols on organoid sections, and measurement of total organoid cell viability, are all also described.

Histological and Histochemical Features of the Newly Lactating Udderinlocal Breed She Goat (Caprice Hirccus)

Six samples of one day post parturition mammary glands of local breed goats were used for this work. The samplesof glands were dissected the tissue specimens were fixed in 10% formalin solution. The specimens were processedaccording to paraffin embedding technique, sectioned at 5-6μm and stained by hematoxylin and eosin, Masson’sTrichrom stains and Verhoff stain. The newly lactating mammary gland was covered by very thick, well glandularskin that measured 210.32±9.03μm. The mammary parenchyma was composed of groups of huge mammarylobes which separated by well vascular thick layer of inter lobar fibrous tissue that measured 986.19±23.10 μm(Thickness). The mean of surface area of mammary lobe was134785.48±15.89μm², it was consisted of numeroussmall size lobules that measured 9848.79±23.75μm² and separated from each other by thin layer of fibro muscularinter lobular connective tissue that measured 138.16±4.11 μm. The lobule was composed of mammary alveoliwhich revealed marked secretory activities. The mean surface area of alveolus was 784.79±19.09 μm² that linedby cuboidal cells that measured 14.11±1.09 μm. Duct system of newly lactating mammary gland was intra lobularduct, large inter lobar duct, largest lactiferous sinus, the lactiferous sinus was drainage to glandular cisterna thatopened into teat cisterna and the later opened into teat canal. Gland cisterna was huge chamber had wide irregularshaped lumen. The mucosa of gland cisterna had few mucosal folds which lined by simple cuboidal epitheliumand supported a lamina propria of dense fibro muscular connective tissue. The epithelium of cisterna which closeto the mammary lobules was supported thick layer of well vascular fibrous connective tissue. Teat cisterna wascomposed of mucosal folds that supported by fibro muscular layer with venous plexus.

Histomorphological and histochemical study of the fallopian tube during pregnancy and post-partum of the cape hare rabbit (Lepus Capensis)

This research was investigate the Anatomical and Histochemical structure of uterine tube during period of pregnancy and postpartum in cap hare rabbits. Eighteen female rabbits were used. The target samples were processing with paraffin embedding technique the tissue sections stained with H&amp;E stain, and Masson's trichrom stain. Results; The length and weight of the uterine tube were (91.31 ± 0.72mm), (0.53 ±0.01g) at pregnancy. While. its (84.76 ±0.22mm), (0.41 ±0.01g) at post-partum. The infundibulum has a funnel-shaped end. Simple mucosal folds are bordered with pseudo stratified columnar epithelium. The lamina propriea-sub-mucosa is made up of connective tissue that is both vascular and cellular. The ampulla was highly vascularized and had a twisted appearance. The mucosal folds of the ampulla were lined by pseudo stratified columnar epithelium, which consisted of three cell types: mucous secretory cells, non-secretory cells and basal cells. The lamina propria was a thin layer of fibrous connective tissue that exposed a significant amount of fibroblast. The isthmus was short and straight at post-partum, and it was held in place by the mesosalpinx. Mucosal folds in the tunica mucosa are mild and short.

Influence of Diabetes on Morphometric Index of Ovarian Follicles in Streptozocin-Induced Rats

The study aimed to determine the effect of diabetes on follicle development by measuring the diameters of rats' ovarian follicles. The structure and function of many organs change by being affected by diabetes. The ovary is an essential organ of the reproductive system affected by diabetes. The size of the ovarian follicles and corpus luteum can also be affected by diabetes. For this reason, body mass, blood glucose level, and rat ovarian follicles, and corpus luteum diameters were measured in this study. Ten healthy female rats were kept as the control group. In the other rat group, experimental diabetes was induced with streptozotocin (STZ)(60 mg/kg). The rats in both groups were killed after 30 days and their ovaries removed. 5-6 µm sections were made using paraffin embedding techniques and stained with hematoxylin-eosin. On the fifth day of STZ administration to rats, the mass loss of rats was 10%, and the diabetogenic index was 330%. Compared with the control group, diameters of diabetic rats significantly decreased the diameter of the primordial, primary and Graafian follicles, and corpus luteum. The decrease in the diameter of secondary follicles of diabetic rats was not significant. The percentage shrinkage index was the highest in the corpus luteum with 37%. As a result, it can be said that diabetes influences the size of the ovarian follicles and especially the corpus luteum, thereby negatively affecting the ability to improve oocyte quality. Diabetes-related follicle diameter may shrink and cause infertility. It may be essential to measure the diameters of the follicles in vitro fertilization studies in patients with diabetes.

Polycystic Kidney in Adult Guinea Pig: A Case Report

Polycystic kidney disease (PKD) is a genetic or acquired disorder characterized by progressive distension of multiple tubular segments, manifested by fluid accumulation, and growth of non-neoplastic epithelial cells. A male, 2.5 years old, Dunkin Hartley guinea pig, which was kept as a retired breeder in a lab animal facility, TANUVAS, was found dead and subjected for necropsy. History of dullness, depression, anorexia and emaciation was reported. Grossly, both the kidneys were enlarged with multiple, variable sized, spherical transparent cysts with clear fluid. The gall bladder was slightly enlarged with thick bile and a few small, black coloured stones. The tissue samples were fixed in 10% neutral buffered formalin and processed through paraffin embedding technique. Histopathological examination of kidney revealed multiple, variable sized cysts mostly in the cortical areas and the cyst wall was lined by flattened epithelium. Partial to complete loss of glomerular tufts, glomerulosclerosis, tubular epithelial cells degeneration and necrosis were also observed. Some cysts were filled with eosinophilic fluid. Interstitial areas revealed the presence of mononuclear cells infiltration. The basement membrane which supported the cyst wall showed mild thickening by special staining like Masson’s trichrome and Periodic Acid Schiff. Liver showed multifocal, periportal fibrosis with mononuclear cells infiltration and bile duct hyperplasia. Lungs revealed pulmonary congestion, emphysema and bronchiolar epithelial cell degeneration with peribronchiolar mononuclear cells infiltration. Presence of multiple cysts with degenerative changes in adjacent tubules and glomeruli of both the kidneys might have lead to the death of the animal.

Cellular and Metabolite Changes in the Secondary Phloem of Chinese Fir (Cuninghamia lanceolata (Lamb.) Hook.) during Dormancy Release

Wood in the cold temperate zone is the product of the alternation of the growing season and the dormant period of trees, but our knowledge of the process of dormancy release in trees remains limited. Chinese fir (Cuninghamia lanceolata (Lamb.) Hook.) was used to investigate cellular and metabolite changes in the secondary phloem tissue during dormancy release. The sampling dates were 2 March, 28 March, and 13 April. The microsections of wood-forming tissue were prepared using the paraffin embedding technique to observe the formation of cambium cells; metabolites in secondary phloem cells were extracted using a methanol/chloroform organic solvent system. The results showed that the secondary phloem consists of phloem fibers, sieve cells and phloem parenchyma. The cells were regularly arranged in continuous tangential bands and were in the order of Phloem fiber-Sieve cell-Phloem parenchyma-Sieve cell-Phloem parenchyma-Sieve cell-Phloem parenchyma-Sieve cell-Sieve cell-Phloem parenchyma-. The Chinese fir cambium was in dormancy on 2 March and 28 March, while on 13 April, it was already in the active stage and two layers of xylem cells with several layers of phloem cells were newly formed. The width of the cambium zone increased from 18.7 ± 5.7 μm to 76.5 ± 3.0 μm and the average radial diameter of sieve cells expanded from 15.4 ± 7.5 μm to 21.5 ± 7.4 μm after dormancy release. The cambium zone width and the average radial diameter of sieve cells before and after dormancy release were significantly different (p &lt; 0.01). The phloem parenchyma cells without resin were squeezed and deformed by the sieve cells, and the width of the phloem during the active period was 197.0 ± 8.5 μm, which was larger than that during the dormant period. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS)-based metabolomics was employed to analyze the secondary phloem of Chinese fir on 28 March and 13 April. Thirty-nine differential metabolites during dormancy release were detected. The results showed that the composition of Chinese fir metabolites was different before and after dormancy release. The relative increase in pyruvic acid and ascorbic acid contents proved that the rate of energy metabolism in Chinese fir increased substantially after dormancy release. Changes in cell development and the composition of metabolites revealed that the dormancy release of Chinese fir was at early April and the formation period of phloem tissue is earlier than xylem tissue.

Histology and structure of the testicles in three species of Atelopus frogs (Anura: Bufonidae) endemic to the Sierra Nevada de Santa Marta, Colombia

Introduction: Testicular histology constitutes one of the least explored aspects in frogs of the genus Atelopus. This taxonomic group shows an alarming population decline; therefore, its reproductive biology is one of the greatest topics of interest for its conservation. Objective: To describe the testicular morphology and the spermatogenetic lineage cells in adult males of Atelopus laetissimus, Atelopus nahumae, and Atelopus carrikeri in the Sierra Nevada de Santa Marta, Colombia. Methods: During June – July 2017 and 2018, sampling was conducted in the localities of San Lorenzo and Páramo Cebolletas, Sierra Nevada de Santa Marta (SNSM), to collect 15 adult males, 5 per species. Testes samples were fixed in Bouin to be processed by the standard paraffin-embedding technique. Histological sections (3 mμ) were stained with Hematoxylin-eosin and Mallory-Heidenhain-Azan-Gomori's. For the description and photographic register of the germ cells, the photonic microscopy technique was used with the differential interference contrast system. Results: The testes are oval organs, compact, light yellow color, and with little vascularization. Externally, they are surrounded by a thin albuginea tunic constituted by regular dense connective tissue. Inside this layer, they are composed of numerous seminiferous tubules of hexagonal contour, in which germ cell cysts are distinguished at different stages of spermatogenesis (spermatogonia I and II, spermatocyte I and II, and early and late spermatids) and spermiogenesis (spermatozoa in fascicles and free spermatozoa). Separating the seminiferous structures is the interstitial tissue in which Leydig cells and blood vessels stand out. Additionally, in the cranial part of the testis, the Bidder's organ was found, formed by two distinguishable regions, the cortex and the medulla. In the cortex, there are previtellogénic oocytes of different sizes surrounded by a monolayer of flat follicular cells. For its part, the medullary region is the connective tissue that nourishes the oocytes and is constituted by blood capillaries. Conclusions: The gonads of the three species analyzed present a cystic cellular organization similar to other anurans, where all stages of spermatogenesis and spermiogenesis were identified, possibly indicating a continuous reproductive activity. Likewise, the Bidder’s organ is reported for the first time in the three Atelopus species, which allows suggesting a possible sexual reversion in case of a population decrease of females as a reproductive strategy.

Morphological and Histological Study of the Pineal Gland in local breed Goats (Capra hircus)

The current study was carried out to investigate both morphological and histochemical features of pineal gland in goat. Eight samples of pineal glands were used. The morphological study was involved the position, relation, weight, length and width of the glands. The tissue sections were prepared for paraffin embedding technique and stained with Hematoxyline &amp; Eosin, Masson trichrom, Alizarin and PAS stains. The morphological result of pineal showed slightly irregular round-spherical shaped, grayish white in color. It located in the same mid-depression between the thalami body and two colliculi. The weight of gland was 0.478±0.02 mg and measured 2.10±0.06mm in length and 2.52±0.15mm in width. Histologically, the pineal gland has enveloped by a smooth thin layer of loose connective tissue capsule showed less trabiculae that carried the blood vessels into gland and the trabiculae have composed of fine collagen fibers. The glandular parenchyma showed marked lobular pattern of division with marked lighter central region which showed less cellular population. The glandular lobule consisted of three types of cells; Type I pinealocytes or dark pinealocytes, the second type was the less populated type II pinealocytes or light pinealocytes and the third type was the glial cell. The Alizarin stain was revealed multiple of variable size calcium patches within collagen fibers. The cytoplasmic contents of both pinealocytes types I &amp; II showed no glycogen granules.

Investigations into Foot and Mouth Disease Outbreak among Small Ruminants in Karnataka State of India

Background: Foot-and-Mouth Disease (FMD) remains a serious threat to the Indian livestock sector due to significant economic loss associated with it. Systematic vaccination of large ruminants over the years has lead to a gradual reduction in the number of disease outbreaks in India. However exposure to FMDV infection in small ruminants has been recorded during the past few years (Rout et al. 2013). Sheep and goat population have not been vaccinated so far against FMD under the FMD-Control program (FMD-CP). The present study highlighted the outbreak of FMD in small ruminants in Karnataka, India.Methods: During the period 2018-19, seven suspected FMD outbreaks among sheep population in Bellary and Tumakuru districts of Karnataka state were investigated. Tongue epithelium (oral swabs) and foot lesions (n=23) from clinically affected sheep and tissues such as heart, lung, liver, spleen, lymph nodes and kidneys from lambs during post mortem (n=67) were collected. All the samples were processed in the laboratory for the detection of FMD virus antigen by employing Serotype differentiating antigen detection ELISA and by multiplex PCR. Heart tissue samples were also collected in buffered formalin for histopathology study and processed by routine paraffin embedding technique and stained with Hematoxylin and Eosin (H and E). Serum samples from the recovered animals were collected and screened by NSP-ELISA and LPB-ELISA to check the antibody status in the affected herd. Result: A total of seven suspected outbreaks of FMD involving 688 small ruminants was investigated. The outbreak of FMD due to FMDV serotype O was confirmed by ELISA and multiplex PCR assays. Clinically, the affected adult sheep showed typical signs of FMD, while mortality in young lambs was observed without apparent signs of disease. Histologically, heart tissues from FMD affected lambs showed myocardial necrosis with marked aggregations of lymphocytes and neutrophils in the myocardium and perivascular spaces. History of FMD outbreaks in cattle and common grazing land for the livestock, as well as sheep within the reach of these villages, may be the major contributing factors for the outbreaks in sheep populations.

A cystic trichoepithelioma in a dog - A case report

Trichoepithelioma is a benign, well demarcated subcutaneous tumor that originates from hair follicle matrix or inner root sheet. A 4 year-old female Golden Retriever was presented to Department of Veterinary Clinical Complex, Veterinary College, Hassan with a history of growing mass on the left lateral digit of right forelimb. On physical examination, the mass was found to be soft in consistency and radiography revealed soft tissue swelling at 5th digit involving all the 3 phalanges. The mass was surgically excised and representativetissue samples from the mass were collected and processed by routine paraffin embedding technique, sectioned at approximately five micron thickness and stained with routine Haematoxylin and Eosin method. Histo-pathological examination of the mass showed several cysts of variable size with trichogenetic differentiation. Cysts wall were lined by thickened basal lamina cells having palisade arrangement of cells near the periphery along with shadow cells in the cystic lumen. Trichoepithelioma is considered as significant because of its histological similarities to basal cell carcinoma, and although benign, there is a rare risk of malignant transformation. The present communication describes pathological findings in a case of cystic trichoepithelioma involvingthe let lateral digit of right forelimb in a female dog.