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Open Access
Gastro-enteropathy induced by silver nanoparticles in wistar rats
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2022-01) Verma, Jiya; Agarwal, Seema
The present study investigated the functional and pathomorphological effects of silver nanoparticles on gastro- intestinal tract at NOAEL dose in Wistar rats for a period of 90 days. A total of 35 rats of 6-8week age were divided randomly into group Ι (control group) and group II (treatment group). Group II rats were administered silver nanoparticles orally at NOAEL dose (30 mg/kg body weight/ day). Behavioral changes such as dullness and hypoactivity were noticed in most of the rats of Group II, although no overt clinical symptoms were observed. Collectively, these observations revealed that silver nanoparticles have toxic effects as such as gastropathy and enteropathy at NOAEL dose. The observations regarding pathomorphological alterations indicates that the most vital organs for the digestion and absorption of food i.e. stomach and intestine were severely affected. There was a significant decrease in antioxidant activity, increase in lipid peroxidation, increase in apoptosis and decrease in enzyme activity of α- glucosidase, β-glucosidase and β-galactosidase in intestinal homogenate and caecal digesta samples respectively of Group II rats. Grossly, slight congestion was noticed in the stomach as well as intestine. Histopathologically, the glands of stomach were found to be atrophied and damaged while the gastric pits were reduced. The proximal, middle and distal parts of small intestine were found to undergo destruction of mucosal glands, breaking, blunting and fusion of villi was observed along with congestion and infiltration of polymorphonuclear leucocytes. Silver nanoparticles have been observed to be extremely irritating to the lining of gastro-intestinal system. Our results unambiguously demonstrate the inimical effects of silver nanoparticles on gastro-intestinal tract in wistar rats even at NOAEL dose on the stomach and intestine. The present study was of a short duration, so, studies with longer duration and in various animal models are required to assess the effects of silver nanoparticles on health as well as in the environment.
Open Access
Clinocopathological studies on nanosilver administered Wistar rats
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2021-12) Neeraj Kumar; Batra, Munish
The present study investigated the pathological effects of nanosilver at NOAEL dose in Wistar rats for a period of 90 days. A total of 35 rats of 6 weeks age were divided randomly into group 1 (control group) and group II (treatment group). Group 1 rats were administered nanosilver orally at NOAEL dose (30 mg/kg). In group II rats, most of rats were dull and lethargic and there was decrease in feed intake. A significant time dependent decrease in body weight. Blood parameters shows decrease in packed cell volume (PCV), haemoglobin (Hb), and total erythrocyte count (TEC). Morphology of erythrocytes is normocytic hypochromic anaemia with increase central pallor. Erythrocytic indices shows significant changes including MCH and MCH values were significant increased while MCHC shows nonsignificant changes in treated rats as compared to control rats. Other blood parameters like total leucocyte count (TLC) and total thrombocyte count (TTC) shows significant increase in value in treated group as compared to control. In differential leucocyte count, lymphocyte count was significantly decreased but neutrophil showed significant increase in value while eosinophils, monocytes and basophils showed nonsignificant variation in both groups throughout experiment. Other blood parameters like absolute neutrophil count (ANC) showed significant increase in value while absolute lymphocyte (ALC) was significantly decrease in treated group as compared to control. In biochemical profile there was significant increase in serum aspartate aminotransaminase (AST), alanine aminotransaminase (ALT), and cholesterol while other parameters like total serum protein, serum gamma globulin, serum albumin, serum globulin, and serum glucose showed decrease in value in treated group as compared to control. Other biochemical parameters like serum calcium and serum phosphorus showed nonsignificant changes in both groups throughout experiment. Grossly group I rats shows non-significant changes while group II rats show red patches in liver, red discoloration in lungs and mild congestion in heart. Histopathologically, no significant lesions could be in any organ of group I rats while in group II rats revealed congestion, thrombus formation, and degeneration of hepatocytes in liver, thickening of interalvular wall, emphysema, congestion and increase number of mononuclear cells in lungs. kidneys revealed the congestion of glomerular capillaries, interstitial haemorrhage, and coagulative necrosis. Heart of group II rats revealed congestion, necrosis, and mild enema in cardiac muscle fibres. Other organs like , spleen, uterus and testis showed non-significant changes. Collectively, these observations indicated that nanosilver at NOAEL dose has toxic effects on heath including serum biochemicals and tissue changes. The observations regarding pathomorphological alterations indicates that liver, kidneys, and lungs, were the most affected organs in nanosilver toxicity. Mild effect on heart was also observed. Collectively our results suggest that nanosilver have toxic effects even at NOAEL dose on the repeated exposure. The use of silver nanoparticles should be used restricted or avoided.
Open Access
Effect of lactation stages on hematobiochemical entities and milk somatic cell count of indigenous Badri cattle
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2020-10) Thakur, Swati; Huozha, R.
Badri cattle is first registered indigenous cattle breed of Uttarakhand, mainly reared in hilly areas of Kumaon and Garhwal regions. It is a dual purpose breed, well adapted to hilly areas, prevailing climatic conditions and resistant to many diseases. This breed has poor milk yield (0.5 to 2.0 kg/day) with short lactation length (208 ± 20 days), but have excellent drought ability and requires less external inputs. Several factors that affects the normal physiology, out of which lactation and pregnancy period are highly affected. Total forty Badri cows (200±50 kg BW, 1-6th parity) belonging to different lactation stages were selected from the conservation unit maintain at IDF, Nagla GBPUAT, Pantnagar for the present study to investigate their hemato-biochemical entities, osmotic fragility test of erythrocytes and milk somatic cell count. Selected cows were divided into five groups according to their lactation stages as early, early mid, late mid, late lactation stages and another group of non-lactation having eight cows in each group. Both blood samples and milk samples were collected. Hematological entities demonstrated a reverse pattern of values between PCV and ESR during lactation stages. TEC was lowest in late mid lactation while appear highest during early mid and late lactation stages. MCV and MCH were highest during late mid lactation and lowest during early and early mid lactation stages. DLC lymphocytes was highest during early lactation stage. Percent hemolysis of erythrocytes was highest during early mid lactation at 0.9% and 0.5% saline concentration. Serum total protein and albumin were highest during early lactation and decreased as advance in lactation stages while serum creatinine, glucose, cholesterol and triglycerides showed reversed pattern. Serum ALT was highest in early mid lactation and lowest during early and late mid lactation stages. During late lactation serum ALP was highest whereas CK was lowest and during late mid lactation serum ALP was lowest whereas CK was highest. Serum phosphorous during late mid lactation showed lowest while Ca: P ratio was highest. Milk somatic cell count (SCC) was increased during early lactation, late mid lactation and decreased during early mid and late stages of lactation. Thus, concluded that different stages of lactation highly affected the normal physiological state which imposed an alteration in the metabolism of lactating cows.
Open Access
Clinicopathological alterations induced by the combined effect of iron and aluminium nanoparticles in wistar rats
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2020-10) Kohli, Nisha; Agarwal, Seema
The present experiment was designed to know the clinicopathological alterations caused by the combined effect of iron and aluminium nanoparticles in Wistar rats in a repeated dose study for 90 days. The experimental design consisted of 35 Wistar rats randomly divided into 2 groups; GI as control group with 20 rats and GII as treatment group with 15 rats from day 0 and till 90 DPT. Control group was provided with standard feed and RO water and in treated group along with standard feed and RO water, nano-iron oxide and nano-aluminium oxide mixed in distilled water was also gavazed at the dose half of the NOAEL dose viz. 15mg/kg/BW and 3mg/kg/BW respectively. Upon routine examination, rats did not show any abnormal behaviour and clinical sign. However, there was decrease in mean body weight in the treated group. Haematological parameters showed decrease in total erythrocyte count (TEC), packed cell volume (PCV), total leucocyte count (TLC) and absolute lymphocyte count (ALC) and decrease in haemoglobin, mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC) in treated group as compared to control group. Biochemical examination demonstrated increase in aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatinine and BUN whereas there was decrease in serum total protein, serum albumin, serum globulin and serum gamma globulin in treated group as compared to the control group. Immunological parameters demonstrated decrease in HI titer, ELISA, B and T lymphocyte blastogenesis. Histopathology in nano-iron and nano-aluminium treated rats. Liver showed degenerative and necrotic changes in the hepatic cells with hemorrhages, congestion and infiltration of mononuclear cells. Lung revealed emphysema, atelectasis and increase in the thickening of the interalveolar septa alongwith mononuclear cell infilteration. Kidney shows lesions of glomerulonephritis. Intestine revealed increase in the number of the goblet cells, along with the shortening, blunting and fusion of villi. Spleen showed congestion, necrosis and depletion of the white pulp area. Heart muscles showed degeneration and necrosis of muscle fibres. Brain revealed neuronal degeneration, gliosis and perivascular cuffing. Thus the present study suggest that nano iron and nano aluminium at the rate of half of their NOAEL dose i.e. 15mg/kg BW and 3 mg/kg BW when gavaged for 90 days causes pathological alterations in the exposed rats. Though some of the results were not statistically significant but findings from the present investigation confirm the pathological changes on various parts of the body that leads to immunopathy, hepatopathy, nephropathy and neuropathy which may result into the down regulation of immunity. Also, histopathological studies suggested that accumulation of these nanoparticles in body induces structural alterations in the various organs. Further studies, should be carried out in different animal models using varied doses and increased duration of nano iron and nano aluminium to exactly find out the pathological and immunosuppressive properties of these nanoparticles.
Open Access
Immunopathological studies of combined effect of copper and aluminium nanoparticles in wistar rats
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2020-10) Rathor, Charu; Chauhan, R.S.
The present experiment was conducted to study the effect of copper and aluminium oxide nanoparticles on the immune system of Wistar rats. The experimental study was conducted for period of 90 days. Thirty five wistar rats were randomly allocated into two groups i.e. control and test group containing 20 and 15 rats each, respectively. During the entire course of experiment, control rats were received distilled water and standard feed. While test rats were orally gavaged with Copper oxide nanoparticles (CuONPs) and Aluminium oxide nanoparticles (Al2O3NPs) at half the NOAEL dose i.e. 50mg/kg/day and 3mg/kg/day, respectively along with standard feed and water in the morning time for a period of 90 days. Rats were regularly monitored for any abnormal clinical signs and change in behavior during the experiment. Various hematological, biochemical and immunopathological parameters were studied along with pathomorphological alterations in rats at day 0, 30, 60 and 90 of the experiment. During the entire course of the experiment no aggressive behavior and abnormal clinical signs were showed by the test rats when compared with the control rats. Though reduced body weight was observed in test group rats relative to the control group rats. Study of various hematological parameters showed significant decrease in the hemoglobin, TEC, TLC, ALC and mean lymphocyte count and decrease in PCV, MCH and MCHC in the test rats when compared with the control rats. Biochemical parameters exhibited significant reduction in total serum protein, serum albumin, serum globulin and serum gamma globulin and significant elevation in the level of serum creatinine, serum BUN and liver function enzyme i.e., AST and ALT in test rats. Various immunological parameters were studied during the experiment to investigate the effect of copper and aluminium oxide nanoparticles on body’s immune response. Decreased value of mean HI titre and mean ELISA in test group revealed suppressed humoral immunity in test group. In delayed type hypersensitivity reaction (DTH) decrease in the thickness of the DNFB sensitized skin and in macrophage function test decrease in NBT positive cells was indicative of suppression in cell mediated immunity. During the lymphocyte stimulation test, reduction in Δ OD in splenocytes was observed by using Con-A, PHA-M and LPS mitogens in the test rats. The results denote the lymphocytotoxic effect of copper and aluminium nanoparticles against both mature and immature T and B lymphocytes. Histopathological lesions were observed in various organs of test rats. Liver showed congestion in central vein, degenerative and necrotic changes in hepatocytes, kupffer cell proliferation and anisonucleation in hepatocytes. Kidneys showed degeneration and necrosis of tubular epithelium, glomerular atrophy, congestion and haemorrhage in glomeruli and interstitium and mononuclear cells infiltration in glomeruli and interstitial spaces. Lung sections demonstrated congestion, emphysema and atelectasis, thickening of interalveolar septa due to infiltration of mononuclear cells. Spleen sections revealed congestion, necrosis and depletion of lymphoid cells at some places. Brain showed neuronal degeneration, perivascular cuffing, satellitosis and gliosis. Heart sections showed congestion in blood vessels and necrosis. Intestinal sections showed lesions of catarrhal enteritis and infiltration of mononuclear cells. Testis sections revealed gap between the seminiferous tubules, focal loss of germinal epithelium, distorted arrangement of spermatogenic cells and degenerative and necrotic changes in seminiferous tubules. Consequently, the present experimental study suggests that nano-copper and nano-aluminium in combination exhibits deleterious effects on immune system of wistar rats when given at the rate of half of the NOAEL dose. The co-exposure of these nanoparicles cause immunosuppression by damaging the immune cells, alter hematological and biochemical attributes along with destructive pathomorphological changes. Based on the above findings, it can be concluded that nanoparticles in combination induce more pronounced immunopathological alterations as well as immunosuppression even in the short period of the experiment.
Open Access
Pathological studies on nanonickel administered wistar rats
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2020-08) Singh, Jitendra; Batra, Munish
The present study investigated the pathological effects of nanonickel at NOAEL dose inWistar rats for a period of 90 days. A total of 35 rats of 6 weeks age were divided randomly into group Ι (control group) and group Π (treatment group). Group Π rats were administered nanonickel orally at NOAEL dose (5 mg/kg). A significant time dependent decrease in body weight, decrease in PCV, decrease in total erythrocyte count is seen. There were nonsignificant decrease in haemoglobin (Hb) is seen. In Erythrocytic indices MCV shows both significant and nonsignificant changes while MCH and MCHC shows significant increase in values in treated rats as compared to control rats. The total leucocyte count (TLC) decreases significantly in treated rats as compared to control rats. In differential leucocyte count, lymphocyte and neutrophil shows significant decrease while eosinophils shows significant increase in value while monocyte shows nonsignificant variation. In biochemical profile there is nonsignificant increase in glucose value and nonsignificant increase in globulin value, significant decrease in total protein and gamma globulin, and significant increase in serum creatinine, serum aspartate aminotransaminase (AST), alanine aminotransaminase (ALT) is seen in treated group as compared to control group. In T- lymphocyte blastogenesis assay, CON-A, PHA-M and LPS shows significant decrease in value in treated rats as compared to control rats. In response to humoral immunity there were significant decrease in HI titre and ELISA value in treated rats as compared to control rats. Delay type hypersensitivity shows significant variation in reaction at 48 and 72 hr post challenges. Histopathologically, group Ι shows no observable lesion while in group Π there were congestion in liver, hyperplasia of kupffer cells is observed while in lungs there were thickening of interalveolar septa and infiltration of mononuclear cells, emphysema is observed, kidney reveals the necrotic changes in tubules of kidney and loss of bowman’s space and glomerulonephritis. Group Π testis reveals degenerative and necrotic changes in seminiferous tubules along with infiltration of mononuclear cells, there was also degenerative and necrotic changes in sertoli cells. In intestine there were hypertrophy and hyperplasia of goblet cells, Group Π spleen revealed increase in red pulp area and deposition of haemosiderin pigment in spleen of nanonickel treated rats. Collectively, these observations indicated that nanonickel have toxic effects as hepatopathy, nephropathy and immunopathology at NOAEL dose. The observations regarding pathomorphological alterations indicates that most of the vital organs of the body including liver, kidney, heart, lungs, spleen, testis were the most affected organs in nanonickel toxicity. Mild effect on other organs including intestine was also observed. The results did not indicate any apparent gross lesion. However, its effect on immune system after oral administration is inhibitory in nature. Collectively our results suggest that nanonickel have toxic effects even at NOAEL dose on the repeated exposure.
Open Access
Immunopathology of calcium and aluminium nanoparticles in Wistar rats
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2020-08) Deeksha; Chauhan, R.S.
The present experiment was conducted to study the effect of nano-calcium carbonate and nanoaluminum oxide on the immune system of Wistar rats. The experimental study was conducted for period of 90 days. 35 Wistar rats were divided randomly in 2 group viz. control and test. Control group contain 20 and tests group contain 15 experimental animals. From day 0 and till 90 day of the experiment, control group was provided with standard feed and RO water and in test group along with standard feed and RO water, nanocalcium carbonate and nao-aluminium oxide mixed in distilled water was also gavazed at the dose half of the NOAEL dose viz. 500mg/kg body weight and 3mg/kg body weight respectively. All the rats were monitored regularly for any change in behavior and other abnormal clinical sign. Various hematological, biochemical and immunopathological parameters were studied along with pathomorphological changes in rats at 0, 30, 60 and 90 DPT of the experiment. During the entire course of the experiment, rats did not show any abnormal behavior and clinical sign. However decreased body weight was observed in the rats of the test group when compared with control group. Hematological studies also showed significant decrease in TLC and ALC and nonsignificant decrease in ANC, Hb concentration, TEC, and PCV in the test rats as compared to the control rats. Biochemical profile study shows significant decrease in the level of serum total protein, serum albumin and non significant decrease in serum gamma globulin, significant increase in level of serum creatinine, ALT, AST and non significant increase in serum glucose in test group when compared with control. Effect of nano-calcium and nano-aluminium on various immunological parameters was also studied during the experiment. Reduced titre of HI and ELISA test values in NPs treated rats shows the deleterious effect of NPs on the humoral immune response. Decreased thickness of the DNFB applied skin in delayed type hypersensitivity reaction and NBT positive cell in macrophage function test in the test rats shows harmful effect of the nano-calcium carbonate and nano-aluminium oxide on the cell mediated immunity. Decreased optical density during the lymphocyte stimulation test performed in spleenocytes of the test rats using Con-A, PHA-M and LPS mitogens also shows lymphotoxic effect against both mature and immature T lymphocyte and B lymphocyte. Histopathological study was also conducted to study the toxic effect of nano-calcium carbonate and nano-aluminium oxide on various organs. Liver of the test rats when compared with the control rats shows degenerative and necrotic changes in the hepatic cells with hemorrhages, congestion and infiltration of mononuclear cells around the central vein and portal triad. Increase in the thickening of the interalveolar septa was observed in lungs with the infilteration of the inflammatory cells. Emphysema and atelectasis in the alveoli was also observed. Kidney shows lesions of glomerulonephritis. Intestine also shows hypertrophy increase in the number of the goblet cells, along with the shortening, blunting and fusion of villi. Spleen shows deposition of large amount of hemosiderin pigment with the necrosis and depletion of the white pulp area. Heart muscles also show degenerative and necrotic changes. Seminiferous tubules in the testes show degeneration along with the infiltration of the mononuclear cells around the tubules. Brain shows neuronal degeneration with change in shape and size of the neurons, gliosis and perivascular cuffing in some regions of the brain. Thus the present experimental study suggests that calcium and aluminium nanoparticles cause structural and functional toxic effects through cellular toxicity and immunotoxicity as well as altered biochemical attributes in the test animals when given at the rate of half of the NOAEL dose of these nanoparticles. Though some of the results were not statistically significant but most of the findings of the biochemical, hematological, immunological and pathomorphological parameters were indicative of the mild to moderate immunopathological alterations including immunodeficiency even in the short duration of the experiment.

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