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The study was conducted on apparently healthy caprine lungs to evaluate the efficacy of different plastination techniques by gross biometrical and morphological analysis of the formalin fixed and plastinated specimens. The gross biometrical analysis was made on all the formalin fixed lungs and after that the samples were divided as Group I and Group II with 6 animals in each group. In method I, after proper fixation, the specimens were dehydrated in three changes of acetone (two weeks each), then impregnated in two changes of glycerol for 15 days each and cured in muslin cloth containing corn starch. In method II, modified plastination solution was used as impregnation solution. The plastinates prepared by method I were soft to touch, odorless, the elasticity and colour of lung tissue was near normal, whereas the lungs plastinated by method II were hard to touch and their color turned to whitish grey. The biometric data showed that the shrinkage in weight of lungs was 47.99% in method I and 85.46% in method II. The shrinkage in length of apex to base of right lung was 14.36% and that of left lung was 16.18% by method I. In method II, the length from apex to base was reduced to be 35.84% and 34.83 in right and left lungs, respectively. It was concluded from present study that the plastinated glycerine specimens (Method I) showed lesser deviations from their near natural state as compared to the plastinates prepared by Indigenous method (Method II). The shrinkage was more evident in plastinates prepared Indigenous method (Method II) than by glycerine method (Method I). So, it is recommended that glycerine method of plastination should be preferred to any other method of plastination.

Histochemical study were made on trachea of buffalo foetuses  ranging from 4.3 to 44 cm CRL (crown rump length) to elucidate distribution of neutral and acid Mucopolysaccharides. During initial development up to 15.8 cm CRL tracheal eqithelium was strong to moderately positive from neutral mucopolysaccharides. The ground substance of developing cartilage primodria was weakly positive from acid mucopolysaccharide linging was present on the surface of PAS positive epithelia cells. The primordial of tracheal glands were positive for neutral mucopolysachharide content with well developed PAS positive submucosal tracheal glands.

Comparative studies on some histoenzymic entities wereconducted on the trachea and lung tissues of adult Pashmina,Bakerwali and non-descript goats (n=10 each) inhabitingat different altitudes and geo-climatic conditions of Ladakhand Jammu and Kashmir. The study was conducted inDivision of Veterinary Anatomy, Faculty of VeterinaryScience and Animal Husbandry, R S. Pura, Jammu in 2019.A part of the same was also conducted in the Departmentof Veterinary Anatomy and Histology, GADVASU,Ludhiana. Tissues from these organs were subjected forlocalizing certain tissue enzymes, viz. Alkaline phosphatase(AKPase), Glucose-6-phosphate-dehydrogenase (G-6-PDH), Lactic acid dehydrogenase (LDH), Succinic aciddehydrogenase (SDH) and NADPH. The epithelial liningof trachea showed strong reaction to G-6-Pase and LDH inBakerwali goat, while SDH showed strong reaction inPashmina goats. Again, in regard to lung tissues, AKPaseand LDH showed moderate to strong reactions in Pashminagoat, but SDH exhibited strong reactions. In Bakerwaligoats, AKPase and LDH showed moderate to strongreactions, while in non-descript goats, LDH exhibitedmoderate to strong reactions and G6PDH showed intensereactions. This revealed that not much variations wereobserved in regard to localization of certain histoenzymicentities in trachea and lung tissues of three breeds of goatswhich are the inhabitants of different altitudes and geoclimatic conditions.

The present investigation aims to study the karyology of the three most popular dog breeds as well as indigenous local dog. In this study, we identified the most popular dog breeds of the Punjab region which are maintained as companion animals, or for guarding. Metaphase plates were prepared after culturing of lymphocytes isolated from heparinized blood collected from the identified three most popular canine breeds. The isolated lymphocyte cells were cultured for 70-72 h following the cell cycle arrest at metaphase. The G-banding of the chromosomes was done by Giemsa staining through a standard protocol. The most popular three breeds of dog in the sub-tropical region were Labrador, the German Shepherd, and Pug. There were no significant distinguishable differences between the karyotypes of the dog breeds studied. This study gives insight into karyology information, which can be beneficial to the researchers, dog breeders, and kennel clubs. Moreover, it provides information about chromosomal abnormalities which may lead to the study of various fertility, growth, and phenotypic abnormalities problems in dog breeds.

This study was taken up with the objective to assess the effect of limonin on the performance of commercial broilers and quality of meat. Day old chicks (200) were divided into 8 groups, each group contained 4 replicates of 6 chicks each in equal sex ratio. The iso-nitrogenous and iso-caloric diets were fed for 35 days, i.e. starter, grower and finisher phase. Kinnow peel powder (KPP) and solid lipid nanoparticles (SLN) of kinnow peel powder containing 7.47 mg limonin/g was added in the required quantity of feed to supply 0, 0.5, 1.0 and 1.5 mg limonin/bird/day. The data was analyzed using 2×4 factorial design. The data revealed that the birds fed diet supplemented with SLN consumed more feed in comparison to those fed diet supplemented with KPP, resulting in higher gain in weight, but without affecting feed conversion ratio (FCR). The digestibility of CP was lower and that of CF was higher when diet was supplemented with SLN in comparison to the one supplemented with KPP. As compared to control diet, limonin up to 1% level did not have any adverse effect on the digestibility of nutrients, but it was depressed beyond 1% level of limonin supplementation. The limonin beyond 1% depressed the dressing percentage. It was concluded that nano-formulations @ 1.0 mg/bird/d is an effective carrier of limonins, leading to improved growth, health characteristics in broilers and meat enriched with limonin.

The present study aimed to observe the expression of MYCN in lungs of mice following chronic exposure of 2,4-D with or without lipopolysaccharide (LPS). 2,4-D was administered orally dissolved in corn oil at high and low dose (1/10th and 1/20th of LD50) for 90 days. After 90 days of exposure, animals from each group were challenged with LPS/normal saline solution at 80 μg/animal. The lung tissues were processed for microarray and real time studies. LPS resulted decrease (–0.173 fold) in m-RNA expression level of MYCN as compared to control, while High dose of 2,4-D alone and in combination with LPS resulted 0.949-fold change and 1.656-fold change increase in expression of MYCN m-RNA, respectively, as compared to control. Similarly, Low dose of 2,4-D alone or in combination with LPS also altered MYCN expression. The microarray data when validated by Real Time PCR was found to be in concordance with the Real Time PCR data. The data taken together suggest that, high and low exposure of 2,4-D alone or in combination with LPS alters expression of MYCN at m-RNA level.

The poultry sector in India being a commercial activity requires workers to spend more number of hours inside the barns. The exposure of workers to barn air may also increase susceptibility to secondary microbial challenges. However, there are no data on the health effects of exposure to the barn from tropical climates. Because of the central role of the toll like receptor 4 (TLR-4) in endotoxin-induced lung inflammation, we evaluated expression of TLR-4 following single and multiple exposures to poultry barn air followed by lipopolysaccharides (LPS) challenge. Male mice (80) aging 7-8 weeks were divided into 4 groups, viz. 3 treatments and 1 control (20; each). Among the treatment groups, Group 1 was exposed to poultry barn air for single day, group 2 for 6 days (Mon to Sat) and group 3 for 24 days (4 weeks, Mon to Sat) for 8 hrs. At the end of exposure period, half of the animals (10) from each group were challenged with LPS @ 100µg/ animal. Immediately following sacrifice, the lung tissues were collected to study the expression profile of TLR-4 mRNA by quantitative RT- PCR. Single exposure did not show any significant change in the expression of TLR-4 mRNA, while 6 days multiple exposures resulted in almost 6 fold increase in the expression of TLR-4 mRNA. There was no change in the expression of TLR-4 mRNA after 24 days multiple exposures compared to control suggesting that prolong multiple exposures may induce adaptation. However, single and 6 and 24 days multiple exposures followed by LPS challenge showed 6, 47 and 17 fold increase in the expression of TLR-4 mRNA, respectively. The data showed exposure to poultry barn air alters TLR-4 expression which may underlie altered responsiveness to LPS challenge.

In the present study, we used high-throughput sequencing, miRNA-seq, to discover and explore the expression profiles of known and novel miRNAs in TLR ligand-stimulated vis-à-vis non-stimulated (i.e. Control) peripheral blood mononuclear cells (PBMCs) isolated from blood of healthy Murrah buffaloes. Six small RNA (sRNA) libraries were multiplexed in Ion Torrent PI chip and sequenced on Ion Proton System. The reads obtained were aligned to the Bos taurus genome (UMD3.1 assembly), which is phylogenetically closest species to buffalo (Bubalus bubalis). A total of 160 bovine miRNAs were biocomputationally identified in buffalo PBMCs and 130 putatively novel miRNAs (not enlisted in the bovine mirBase) were identified. All of these 290 miRNAs identified across the six treatment and control samples represent the repertoire of novel miRNAs for the buffalo species. The expression profiles of these miRNAs across the samples have been represented by sample dendrogram and heatmap plots. The uniquely expressed miRNAs in each treatment and control groups were identified. A few miRNAs were expressed at very high levels while the majority of them were moderately expressed. The miRNAs bta-miR-103 and -191 were found to be highly abundant and expressed in all the samples. Other abundantly expressed miRNAs include bta-miR-19b, -29b, -15a, -19a, -30d, -30b-5p and members of let family (let 7a-5p, let 7g & let 7f) in LPS and CpG treated PBMCS and bta-miR-191, -103 & -19b in Poly I:C stimulated PBMCs. Only one novel miRNA (bta-miR-11039) out of 130 identified putatively novel miRNAs, was expressed in all the six samples and differentially expressed (>2- fold) miRNAs were identified. Six of the differentially expressed miRNAs across the groups (bta-miR-421, bta-let-7i, bta-miR-138, bta-miR-21-5p, bta-miR-222 and bta-miR-27b) were subsequently confirmed by TaqMan quantitative reverse transcription polymerase chain reaction (qRT-PCR). Furthermore, the target genes of differentially expressed miRNAs were enriched for the roles in innate immunity and TLR signaling pathways. This maiden study on profiling and cataloguing of bubaline miRNAs expressed in TLR-ligand stimulated PBMCs will provide an important reference point for future studies on regulatory roles of miRNAs in immune system of buffaloes.

Abstract Ethion is an organophosphate used commonly in India despite being banned in many other countries. The present study was designed to study the interaction of ethion and lipopolysaccharide (LPS) together on lung after single low dose ethion exposure. Mice (n = 20) were alienated into control and treatment groups (n = 10 each). The treatment group was orally fed ethion (8 mg/kg/animal/day) dissolved in corn oil. The animals (n = 5 each) from both the groups were challenged with 80 μg Escherichia coli lipopolysaccharide (LPS) intranasally and the remaining animals (n = 5 each) were administered normal saline solution after 24 h. Ethion along with LPS induced lung inflammation as indicated by increased neutrophils and total leukocyte count (TLC) in broncheoalveolar lavage fluid. Ethion induced histomorphological alterations in lung as shown by increased pulmonary inflammation score in histopathology. Real time PCR analysis showed that ethion followed by LPS resulted significant (p

Abstract Background Xanthosine treatment has been previously reported to increase mammary stem cell population and milk production in cattle and goats. However, the underlying molecular mechanisms associated with the increase in stem cell population and milk production remain unclear. Methods Primiparous Beetal goats were assigned to the study. Five days post-partum, one mammary gland of each goat was infused with xanthosine (TRT) twice daily (2×) for 3 days consecutively, and the other gland served as a control (CON). Milk samples from the TRT and CON glands were collected on the 10th day after the last xanthosine infusion and the total RNA was isolated from milk fat globules (MEGs). Total RNA in MFGs was mainly derived from the milk epithelial cells (MECs) as evidenced by expression of milk synthesis genes. Significant differentially expressed genes (DEGs) were subjected to Gene Ontology (GO) terms using PANTHER and gene networks were generated using STRING db. Results Preliminary analysis indicated that each individual goat responded to xanthosine treatment differently, with this trend being correlated with specific DEGs within the same animal’s mammary gland. Several pathways are impacted by these DEGs, including cell communication, cell proliferation and anti-microbials. Conclusions This study provides valuable insights into transcriptomic changes in milk producing epithelial cells in response to xanthosine treatment. Further characterization of DEGs identified in this study is likely to delineate the molecular mechanisms of increased milk production and stem or progenitor cell population by the xanthosine treatment.

Deltamethrin is an insecticide used to control harmful agricultural insects that otherwise damage crops and to control vector-borne diseases. Long-term exposure to deltamethrin results in the inflammation of the lungs. The present study elucidates the molecular mechanism underlying the deltamethrin-induced lung damage. The lung samples were extracted from the Swiss albino mice following the treatment of low (2.5 mg/kg) and high (5 mg/kg) doses of deltamethrin. The mRNA expression of TCR, IL-4, and IL-13 showed upregulation, while the expression of NFAT and FOS was downregulated following a low dose of deltamethrin. Moreover, the expression of TCR was downregulated with the exposure of a high dose of deltamethrin. Furthermore, the immunohistochemistry data confirmed the pattern of protein expression for TCR, FOS, IL-4, and IL-13 following a low dose of deltamethrin exposure. However, no change was seen in the TCR, NFAT, FOS, JUN, IL-4, and IL-13 immunopositive cells of the high-dose treatment group. Also, ELISA results showed increased expression of IL-13 in the BAL fluid of animals exposed to low doses of deltamethrin. Overall, the present study showed that deltamethrin exposure induces lung damage and immune dysregulation via dysregulating the NFAT signalling pathway.

Abstract Background Pesticide residues in food and environment along with airborne contaminants such as endotoxins pose health risk. Although herbicide 2,4-Dichlorophenoxyacetic acid (2,4-D) has been associated with increased risk of lung cancers such as small cell lung cancer (SCLC) among agricultural workers, there are no data on the SCLC signaling pathway upon 2,4-D exposure without LPS or in combination with endotoxin. Methods We exposed Swiss albino mice (N = 48) orally to high (9.58 mg kg− 1) and low (5.12 mg kg− 1) dosages of 2,4-D dissolved in corn oil for 90 days followed by E. coli lipopolysaccharide (LPS) or normal saline solution (80 μl/animal). Lung samples and broncho-alveolar fluid (BALF) were subjected to Total histological score (THS) and total leucocyte count (TLC) and differential leucocytes count (DLC) analyses, respectively. We used microarray and bioinformatics tools for transcriptomic analyses and differentially expressed genes were analyzed to predict the top canonical pathways followed by validation of selected genes by qRT-PCR and immunohistochemistry. Results Total histological score (THS) along with BALF analyses showed lung inflammation following long term dietary exposure to high or low doses of 2,4-D individually or in combination with LPS. Microarray analysis revealed exposure to high dose of 2,4-D without or with LPS upregulated 2178 and 2142 and downregulated 1965 and 1719 genes, respectively (p

Abstract Background Pesticide residues in food and environment along with airborne contaminants like endotoxins pose a tangible threat to living beings. Fipronil, a broad-spectrum insecticide at acute doses in combination with endotoxin has been linked to airway inflammation. Wnt/PCP pathway plays important role in the pathogenesis of chronic inflammatory conditions by contributing to the release of downstream cytokines. However, there is no data on the PCP signaling pathway during fipronil-induced lung damage. This study aims to understand the molecular mechanisms underlying the pulmonary toxicity induced by fipronil alone or in combination with endotoxin. Methods We used a microarray approach to obtain a global view of the transcriptional responses of the lungs exposed to fipronil and endotoxin in a mice model. Lung tissues were harvested from male Swiss albino mice (n=42) following long term oral exposure to high and low doses of fipronil alone or in combination with endotoxin. The differentially expressed genes were analyzed by Ingenuity pathway analysis software to predict the top canonical pathways. The microarray data for selected genes was validated using quantitative PCR and immunohistochemistry. Results Both BAL fluid and histopathology analysis suggested lung damage and altered transcriptomic profile of lung following exposure to high and low dose of fipronil with or without endotoxin. PCP pathway was among the top upregulated canonical pathways following exposure to both doses of fipronil individually or with LPS. Low dose of fipronil increased the expression of WNT-6, MAPK-8, IL-4 and IL-17C while higher dose decreased the expression of MAPK-8 and IL-17C. The expression data verified by realtime PCR was in concordance with microarray data. Conclusion Long-term exposure to low (4.75 mg kg-1) and high (9.50 mg kg-1) dose of fipronil alone or in combination with endotoxins alters the histoarchitecture and transcriptome profile of lungs with the involvement of the PCP pathway. PCP pathway showed higher enrichment in low dose group of fipronil alone or in combination with LPS. The data from the study provides the insights of the potential damage on lungs caused by fipronil and endotoxin interaction and helps to better understand the mechanism of this complex relation.

Background and Aim: Newcastle disease (ND) is considered one of the most important poultry diseases with chicken morbidity and mortality rates up to 100%. Current vaccination programs allow the use of live attenuated vaccines in the field to protect against the disease, which alone is inefficient and requires repeat booster doses. Toll-like receptor agonists (e.g., lipopolysaccharide [LPS]) as adjuvants are the ones, most extensively studied and have shown to be very promising in delivering a robust balanced immune response. In the present study, we have evaluated the potential of LPS to elicit a strong immune response with respect to the elicitation of both Th1 (cell-mediated) and Th2 (humoral) immune arms. Materials and Methods: A total of 72 apparently healthy 1-day-old indigenous unvaccinated chicks were randomly divided into six experimental Groups A to F (n=12). At 8-week of age chicks in Group A, C, and E were vaccinated with live attenuated La Sota strain ND vaccine along with LPS, bovine serum albumin, and normal saline solution, respectively, and those in Group B, D, and E were kept separately without vaccination. Sampling was done on days 0, 1, 3, 7, 14, 21, 35, and 60 after vaccination. After vaccination and respective adjuvant application, Th1 and Th2 cytokine expression were measured in mRNA of both blood and tissue samples. Results: The results were validated by, hemagglutination inhibition and enzyme-linked immunosorbent assay tests, to check for the humoral as well as cell-mediated immune response in blood serum levels. The results showed an increase in mRNA expression of the Th1 biased cytokines in Group A (LPS+NDV) as compared to the control groups. Similar mRNA expression pattern was seen in blood as well as tissue samples. Validation of results also indicates an increase in Cell-mediated Immunity as well as a humoral immune response in Group A (LPS+NDV). Conclusion: The results of the study provided enough evidence to consider LPS as a potential vaccine adjuvants candidate against ND in chicken.

Aim: Chronic exposure to indoxacarb and pulmonary expression of toll-like receptor 9 (TLR-9) in mice. Materials and Methods: In this study, healthy male Swiss albino mice (n=30) aging 8-10 weeks were used to evaluate TLR-9 expression in lungs of mice following indoxacarb exposure with and without lipopolysaccharide (LPS). Indoxacarb was administered orally dissolved in groundnut oil at 4 and 2 mg/kg/day for 90 days. On day 91, five animals from each group were challenged with LPS/normal saline solution at 80 μg/animal. The lung tissues were processed for real time and immunohistochemical studies. Results: LPS resulted increase in fold change m-RNA expression level of TLR-9 as compare to control, while indoxacarb (4 mg/kg) alone and in combination with LPS resulted 16.21-fold change and 29.4-fold change increase in expression of TLR-9 m-RNA, respectively, as compared to control. Similarly, indoxacarb (2 mg/kg) alone or in combination with LPS also altered TLR-9 expression. Further at protein level control group showed minimal expression of TLR-9 in lungs as compare to other groups, however, LPS group showed intense positive staining in bronchial epithelium as well as in alveolar septal cells. Indoxacarb at both doses individually showed strong immuno-positive reaction as compare to control, however when combined with LPS resulted intense staining in airway epithelium as compare to control. Conclusion: Chronic oral administration of indoxacarb for 90 days (4 and 2 mg/kg) alters expression of TLR-9 at m-RNA and protein level and co-exposure with LPS exhibited synergistic effect.