<div>The staining of free hair samples is considered as one of the biggest problem by the Histologists. This technique has been developed for processing and routine staining of free hair samples without the skin. It is based on paraffin embedded blocks of hair samples. The blocks were cut in the rotary microtome at 6 &micro; thickness. After obtaining the sections of 6 &micro; thickness in rotary microtome, the sections were not required to be put in water bath as the hair samples were hard and did not get folded. Further, if water bath was used, this could result in the loss of the hair part from the section. The cuticle did not take any color in the routine staining as it was composed of enucleated or keratinized cells. The cortex had taken the pink colour of eosin which might be due to the composition of cortex with flattened cornified cells and numerous air spaces. The medulla showed a mixture of pink and blue color on staining with Hematoxylin and Eosin solutions.</div>

Technique for Processing and Routing Staining of Hair Samples for Histological Study

Out of 428 faecal 178 (41.59%) animals were found positive for Cryptosporidium parvum oocysts in the bovine faeces in modified Ziehl-Neelsen (mZN) staining and/ or commercial plate and dipstick- enzyme linked immunosorbent assay (ELISA) kits (Cypress diagnostics, Langdorp, Belgium) with statistically non-significant difference in the occurrence rate in cattle and buffalo calves. Seasonal prevalence difference was statistically significant in cattle calves while non-significant in buffalo calves, respectively. The highest overall prevalence was recorded during rainy season (45.15%) and almost same per cent during winter and summer. Prevalence of 67.26, 37.11, 30 and 17.65% was recorded in calves aged below 1 month, 1-3 months, 4-8 months and 9-12 months, respectively (p&lt;0.05). There was no significant difference in the age group prevalence between cattle and buffalo calves. Both sexes of bovines are equally susceptible to the cryptosporidiosis (p&gt;0.05). The prevalence of cryptosporidiosis was 59.54 and 29.41% for diarrhoeic and non-diarrhoeic samples, respectively (p&lt;0.05). Both diarrhoeic and non-diarrhoeic groups of calves aged between 1-30 days were equally susceptible to the infection of Cryptosporidium spp. Almost same per cent prevalence of the infection was observed in dairy calves reared in organized (40.76%) and unorganized farms (42.21%) and the difference was non-significant in both cattle and buffalo calves. Highest prevalence of cryptosporidiosis was found in HF cross calves. The tests failed to detect the oocysts in infected soil samples. There was highly significant difference in the prevalence of cryptosporidiosis estimated by plate/ dipstick-ELISA and mZN staining with highest sensitivity and specificity in plate-ELISA.&nbsp;

Comparative evaluation of conventional staining method and enzyme linked immunosorbent assay kits for the detection of bovine cryptosporidiosis 
&nbsp;

<div style="text-align: justify;">Mammary gland tumors are the most commonly diagnosed tumors in domestic dogs. Although immunohistochemical methods provide valuable information such as the location and semi-quantitative data of the interested antigens in particular tumors, conventional methods like histopathological diagnosis remain useful and necessary for identification and classification of tumors. In the present study, we combined histopathology with immunohistochemical staining for progesterone receptor (PR) and special staining argyrophilic nucleolar organizer regions (AgNOR) for cell proliferation in canine mammary gland tumors. Twenty-nine dogs with primary mammary gland neoplasms (11 benign and 18 malignant) were included in this study. All 29 tumors were stained with AgNOR but only 15 cases expressed positive PR signals in the nuclei of neoplastic alveolar and ductal epithelial cells. A weak positive correlation (r = 0.2) was observed between PR and AgNOR index of benign tumors and a very weak negative correlation (r = -0.1833) between PR and AgNOR index of malignant tumors suggesting that higher cellular proliferation is associated with low expression of progesterone receptors.&nbsp;</div>

<div style="text-align: justify;">A Study on Immunohistochemical Expression of Progesterone Receptor and Cell Proliferation by AgNOR in Canine Mammary Tumors (CMT)</div>

<div>Background: Cryptosporidiosis is an emerging zoonotic protozoan disease caused by Cryptosporidium spp. The infection was reported worldwide from domestic animals and humans, including wild animals. From India, no such reports were published on Cryptosporidium infection in captive wildlife. Hence, a pilot study was conducted to report the occurrence of Cryptosporidium infection in captive wildlife of India.&nbsp; 
 
Methods: Faecal samples (n=788) were collected from 127 captive wildlife species of three zoological parks viz., Sri Venkateswara Zoological Park (SVZP), Tirupati (n=242); Indira Gandhi Zoological Park (IGZP), Visakhapatnam (n=218); Nehru Zoological Park (NZP), Hyderabad (n=328) and screened for Cryptosporidium infection. Preliminary screening of faecal samples was done for the detection of Cryptosporidium oocysts by modified Ziehl-Neelsen (mZN) staining method and the test positives were confirmed by nested PCR targeting 18S rRNA gene. Nested PCR amplicons were sequenced for determining the Cryptosporidium species. The resultant data were statistically analyzed by Fisher/Chi square, Fisher Exact test using SPSS software v 17.0. 
 
Result: In mZN staining method, 7.23 percent of isolates were found to be positive for Cryptosporidium and the highest rate of infection was detected in wildlife at NZP, Hyderabad (8.23%), followed by SVZP, Tirupati (7.44%) and IGZP, Visakhapatnam (5.50%). Cryptosporidium positive faecal samples by mZN staining were further confirmed by nested PCR and positive amplicons were sequenced for determination of Cryptosporidium species. Genetic characterization revealed five species viz., Cryptosporidium parvum; C. ryanae, C. suis, C. muris and Cryptosporidium avian genotype III. The study conclude that, Cryptosporidium infection was prevalent in the captive wildlife from the zoological parks of India and species variation was marked among the wildlife. Based on the available literature, the current study is the first of its kind on the prevalence of Cryptosporidium in captive wildlife from India.</div>

<div>Isolation and Genetic Characterization of Cryptosporidium from Captive Wildlife of India</div>

<div>Background: The cytochemistry and cytoenzymatic profile of the blood cells is an important tool in the hands of clinical veterinary practitioners. It has also applications in various fields of veterinary sciences. Therefore based on its clinical importance present study was conducted on the blood cells of native cattle (Zobawng) of Mizoram to examine the cytochemical and cytoenzymatic characteristics. 
 
Methods: For this study, a total of 12 numbers of blood samples (5 ml) were taken aseptically from adult local cattle of Mizoram, irrespective of sex. The blood smears were prepared and stained with acid ferrocyanide (ACF) for the detection of non-heame iron, toluidine blue (TBO) stain for acid mucopolysaccharides, Periodic Acid-Schiff (PAS) stain for observation of glycogen and Sudan black B (SBB) stain for demonstration of lipid. Acid phosphatase (ACP), alkaline phosphatase (ALP), peroxidase (POX), arylsulphatase (ARS), cytochrome oxidase (CYO), beta-glucuronidase (GUSB) and succinate dehydrogenase (SDH) enzymes were used for cytoenzymatic features of blood cells. The smears were observed under 1000X magnification in an Olympus Trinocular Research microscope. 
 
Result: In the cytochemical studies, basophils showed a positive reaction for mucopolysaccharides in the toluidine blue stain and neutrophils, eosinophils and basophils recorded positive for glycogen in the PAS stain. Eosinophils and basophils also displayed positive for lipids in the SBB stain. The cytoenzymatic studies of eosinophils and basophils recorded positive reactions for ACP, ALP, POX, ARS and CYO activity.</div>

Diagnostic Importance of Cytochemical and Cytoenzymatic Patterns of Peripheral Blood Cells in Native Cattle (Zobawng) of Mizoram, India

Background: In recent years, relevant studies had shown that a large number of adipokines were involved in energy homeostasis in the body, regulating the balance of glucose and lipid metabolism. Resistin was a special type of adipose factor and its discovery and related studies had shown a close relationship between resistin and heart disease. However, there was relatively little research on the structure of heart tissue, so we conducted this present study.&nbsp; 
 
Methods: In order to study the effect of resistin on the heart structure of mice, 20 healthy C57BL/6 mice were randomly divided into experimental and control groups(n=10). The mice of the experimental group were injected with 20 &igrave;l/d of resistin solution through the tail vein. The mice of the control group were injected with the same volume of double distilled water. HandE staining, Wheat Germ Agglutinin (WGA) staining, Masson&rsquo;s trichrome staining and electron microscopy were used to observe the changes of microscopic structure and ultrastructure of mouse cardiomyocytes.&nbsp; 
 
Result: The results of HandE staining showed that the myocardial cells in the experimental group were disordered, the cytoplasm was loosely distributed and the gap between adjacent cells increased. WGA staining showed that the fluorescence green intensity of the mice in the experimental group was higherthan that of the control group. The myocardial cell membrane thickened, the average cross-sectional area of cardiomyocytes was larger than that of the control group. The results of the Masson staining showed that the staining of the cytoplasm of myocardial cells in the experimental group was significantly shallow and the blue structure was significantly increased. The electron microscopy showed that the number of mitochondria decreased and the mitochondria lightly dissolved or disappear. This study confirmed that resistin could change the content of collagen fibers and myofibrils in myocardial cells, induce cardiac hypertrophy and cause certain damage to mitochondria of cardiomyocytes. It was speculated that it could affect the physiological function of cardiomyocytes to some extent.

Effect of Resistin on the Heart Structure of Mouse

Background: A cytochemical and cytoenzymatic study involves using specialized stains to microscopically examine cellular components, including lipids and carbohydrates. 
&nbsp; 
Methods: Freshly prepared blood smears from 26 apparently healthy adult ducks were used for cytochemical and cytoenzymatic analysis. The smears were stained with specific and special cytochemical stains, Periodic Acid-Schiff (PAS) for carbohydrates, Sudan Black B for lipids, acid ferrocyanide for non-heme iron and toluidine blue for mucopolysaccharides, as well as cytoenzymatic stains such as b-naphthol acetate esterase, acid phosphatase, alkaline phosphatase, b-glucuronidase and peroxidase to assess enzymatic activity. 
&nbsp; 
Result: In the White Pekin duck, rare acid ferrocyanide-positive erythrocytes exhibited blue siderocyte granules with cell deformities, while leukocytes and thrombocytes were negative. Basophils strongly reacted to mucopolysaccharides, showing intense violet granules under toluidine blue, whereas other cells were non-reactive. Heterophils exhibited strong PAS positivity for carbohydrates, with eosinophils, erythrocytes and thrombocytes showing moderate positivity. Sudan black B staining revealed strong lipid positivity in eosinophils and heterophils, while other cells were showed negative for same. Cytoenzymatic analysis showed strong acid phosphatase activity in basophils, moderate activity in heterophils and weak activity in eosinophils and lymphocytes. Eosinophils and heterophils also strongly reacted to b-naphthol acetate esterase, with lymphocytes showing weak positivity. Thrombocytes were strongly positive for peroxidase and eosinophils and erythrocytes showed weak activity. Other cells remained non-reactive to b-glucuronidase, alkaline phosphatase and other enzymes, indicating distinct staining patterns across cell types.

Cytochemical and Cytoenzymatic Architectural Properties of the Blood Cells of White Pekin Duck

This study documented macroscopic and microscopic features and mineralization of the caudally located tracheal rings and syrinx in two ostrich (struthio camelus) having three years of age. The syrinx and trachea of the birds were stained in toto with alcian blue and alizarin red for cartilage and mineralization. Observations on the syrinx and trachea, measurements and photography were performed under stereo-microscopy. They were stained grossly using alizarin red and alcian blue to visualize mineralization and histologically by hemotoxylin and eosin (H&amp;E) to detect ossification areas, if any. Results revealed incomplete tracheal rings located caudally in-between the intact ones. Alizarin red and alcian blue staining displayed mineralized regions grossly. Histological slides by H&amp;E staining showed no ossification. Overall results proposed that alizarin red and alcian blue double staining is a good toll to determine mineralization which is calcium accumulation in the tissue before the formation of bone cells.

Mineralization in the syrinx and caudal tracheal rings in the ostrich

<div>Background: Canine oral fibrosarcomas are locally invasive malignant mesenchymal tumors and the third most frequently occurring neoplasia of oral cavity of dogs. The present study was conducted to record pathological features of oral fibrosarcomas in canines.&nbsp; 
 
Methods: During the study period (September 2020- April 2021), a total of 18 cases of canine oral neoplasia were reported and amongst these fibrosarcoma was diagnosed in five animals (27.78%). The blood samples were collected from all the dogs and the excised tumor mass was processed at Department of Veterinary Pathology, Co.V.Sc. and A.H., Jabalpur.&nbsp;&nbsp; 
 
Result: The hematological profile of all the five dogs showed no variation however; serum biochemical analysis revealed hypercalcemia. The serum MDA concentration was elevated in dogs with fibrosarcoma, indicating oxidative stress. The gross morphology of the tumors included red to pink colored irregular masses, hard in consistency and firmly attached to the underlying tissue. Cytologically, the impression smears revealed spindle shaped cells with round to elongated nuclei along with strands of collagen fibres. Upon histopathological examination proliferation of fibrous connective tissue was observed in the sub-mucosa. Higher magnification revealed several immature spindle shaped fibroblasts along with scanty amounts of collagenous matrix. Masson&rsquo;s trichome staining also exhibited blue stained collagen deposition. Mitotic figures were numerous (8.4 mitosis/10 hpf). The tissue sections stained by modified silver colloid staining were examined at 1000 x magnification. Mean Ag-NOR count of the silver nitrate stained tissue sections was enumerated to be 9.15 dots/nucleus. Immunohistochemical examination detected CD31 immunopositivity in the tissue sections as brown staining of the infiltrating blood vessels, indicating neovascularization of the tumor.</div>

Oral Fibrosarcoma in Dog: Cytological, Histopathological and Immunohistochemical Diagnosis

Background: Sperm quality plays an important role in the animal industry. The study of the sperm quality of Congjiang Xiang pigs is limited and the Rh123/PI double staining has not been used to detect the sperm quality of boars. Thus, the study aimed to evaluate the sperm quality of the Congjiang Xiang pigs and the feasibility of the flow cytometry (FCM) method in assessing sperm quality in pigs. 
 
Methods: Semen samples were evaluated for routine indicators of semen quality and using FCM with single staining and double staining to analyze the integrity of membrane and acrosome, the mitochondrial function. The sperm quality of the Congjiang Xiang pigs was compared with the Large White pigs. 
 
Result: The semen of the Congjiang Xiang pigs had a lower sperm concentration than that of the Large White pigs (P&lt;0.01). There were no significant differences in sperm vitality, sperm motility and abnormality between the two breeds (P&gt;0.05). Single sperm staining revealed a lower sperm acrosomal integrity in the semen of the Congjiang Xiang pigs than that of the Large White pigs (P&lt;0.01). No significant difference between the two breeds in membrane integrity and mitochondrial function was observed (P&gt;0.05). Rh123/PI double staining demonstrated that the rate of live sperm with a normal mitochondrial function of the Congjiang Xiang pigs was significantly higher than that of the Large White pigs (P&lt;0.05). The rate of necrotic sperms in the semen of the Congjiang Xiang pigs was significantly lower than that in the semen of the Large White pigs (P&lt;0.05). Correlation analysis revealed a positive association between the rates of live sperm in the semen of the congjiang xiang pigs and their sperm motility (P=0.051). Our results suggest that Rh123/PI double staining with flow cytometry can provide more reliable information to assess the boar sperm quality.