Sepsis, a systemic inflammatory response syndrome against microbial infection, causes multiple organ failure resulting in death worldwide in human and animals. Vascular hyporeactivity resulting in reduced peripheral resistance is considered to be the leading cause of death in sepsis. Ubiquitylation is a post-translational modification that regulate cellular protein turnover, however, dysregulation of the ubiquitin proteosome system (UPS) alters the protein homeostasis and thereby contribute to various disease conditions. Cullin ring ubiquitin ligase (CRL) is one of the most studied UPS in the context of cancer biology, albeit, its role in infection and inflammation has recently become an exciting area of research. Thus, present study was aimed to evaluate the role of CRL in regulating vascular contractile protein homeostasis and thereby regulating sepsis-induced vascular hyporeactivity. Further, an attempt was made to delineate the efficacy of cullin neddylation inhibitor on the outcome of sepsis, if any.
Present work was undertaken to study the effect of doxycycline and doramectin inmale rats. Following exposure to doxycycline hyclate orally for 28 days at dose rate of 10,16.50 and 33 mg/kg, rats of any doxycycline treated groups did not show any clinicalsigns, mortality or abnormal behaviour. Percent body weight gain was significantlyreduced in rats of group treated with doxycycline (16.50 and 33 mg/kg). A significantincrease in relative weight of left and right testes was found in rats exposed to doxycycline(33 mg/kg). Doxycycline did not produce any change in haematological parameters in ratsof doxycycline treated groups. Blood glucose level, total cholesterol, LDL cholesterol andBUN levels was significantly increased in rats of group exposed to doxycycline (33mg/kg). Rats of group treated with doxycycline (16.50 mg/ kg) exhibited a significantdecrease in blood HDL cholesterol level. Hepatic SOD level was significantly increased inrats of group treated with doxycycline (33 mg/kg).
Present study was undertaken to investigate the sub-chronic effects of chromium (VI) inmale rats using potassium dichromate (0.625 mg/kg, 1.25 mg/ kg, and 2.5 mg/kg)following oral administration. No discernible clinical signs of toxicity were observed atany dose, and weekly average body weight gain remained almost consistent. Both absoluteand relative organ weights exhibited no significant differences among treatment groups.Hematological parameters showed no appreciable variations across the different treatmentgroups. Potassium dichromate exposure demonstrated limited impact on plasma totalprotein, albumin, and globulin levels. While HDL cholesterol exhibited a non-significantdecrease, plasma glucose, cholesterol, and LDL cholesterol levels displayed a slight andnon-significant increase. Non-significant elevations were noted in plasma AST, ALT,creatinine, and uric acid across all treated groups, with minimal changes in total bilirubinand BUN. Hepatic MDA levels increased non-significantly in a dose-dependent manner,accompanied by a non-significant decrease in SOD, CAT, GSH, and GST levels.Testicular MDA increased non-significantly in a dose-dependent manner, with aconcomitant non-significant decrease in SOD, CAT, and GST levels. Testicular GSHlevels showed minimal alteration. A non-significant rise in ACP, LDH, GGT, and FSHlevels in the testis was observed across all treated groups, while testicular SDH andtestosterone levels exhibited non-significant decreases. Necropsy revealed no grosschanges in various organs, including lungs, liver, kidneys, spleen, testes, seminal vesicles,and prostate.
Wound healing is defined as any breach in the continuity of skin. Regenerative pharmacology aims to develop new therapeutic module to hasten the healing. Present study was aimed to evaluate the wound healing efficacy of goat milk based ointment vis a vis its molecular pharmacodynamics in full thickness excisional rat model. 96 healthy male Wistar rats were equally and randomly divided into four groups i.e. group I served as Healthy control/Sham Operated, group II served as silver sulphadiazine treated group and group III and IV served as group treated with 0.5 % and 1% goat milk based ointment, respectively. Square shaped wound of ≈ 2×2 cm2 (400 mm2) was surgically and aseptically created on the back of animals till the depth of panniculus carnosus. Animals were examined routinely till 21 day of the experimental study and healing potential of goat milk based ointment was ascertained by planimetric analysis viz. digital photography, wound area measurement, wound contraction percentage and biochemical assay using hydroxyproline, total protein and DNA content at different intervals i.e. on days 3,7,14 and 21 of the study.
Endometritis is one of the leading causes of reproductive inefficiency, infertility and decreased milk yield in high yielding dairy cows and buffaloes as well as in other species of animals and are associate with severe economic loss. Indiscriminate use of antimicrobial agents has resulted in development of resistance to the synthetic antimicrobials. Plant-derived active principles have promising potential to act as an alternative to conventional antimicrobial agents. In the present study, an attempt was made to investigate the mechanism of action of Prosopis juliflora leaves (PJ) extract against clinical isolates of E. coli.. Out of 42 clinical samples of uterine discharges and 23 isolates from cattle and buffaloes having history of uterine infection, we identified two isolates as virulent E. coli based on the cultural, biochemical and genotypic characterization. Ethanolic PJ extract showed presence of large quantity of total phenolic acid and flavonoid contents as well as promising in vitro antioxidant and radical scavenging activity. PJ extract exhibited a marked in vitro antibacterial effect against S4 isolate and reference strain (ATCC 25922) as evidenced by agar well diffusion test with MIC value of 0.39 mg/ml. This antibacterial action of PJ extract was shown to initiate at 6 h post-exposure while the complete bactericidal action was achieved at 12 h post-exposure.
Present study was undertaken to evaluate the ameliorative effect of α-tocopherol(100 mg/kg) against cartap (16.25 mg/kg and 32.5 mg/kg) induced toxicity following 28days oral exposure in adult male Wistar rats. Cartap exposure did not produce any clinicalsigns of toxicity in rats at both the doses. Weekly Average body weight gain wassignificantly reduced following cartap (32.5 mg/kg). Both absolute and relative organweights did not differ significantly between different treatment groups. Hematologicalparameters did not differ significantly in any of the treatment groups. Plasma Glucose,Total Cholesterol, Triglycerides, AST, ALT, BUN levels were found to be markedlyincreased in cartap alone treatment groups. Plasma calcium levels showed irregular butnon-significant change in response to cartap doses. α-tocopherol co-treatment with cartapproduced non-significant decrease in the plasma levels of Glucose, AST and ALT but thelevels of Total Cholesterol Triglycerides and Calcium were found to be significantlydecreased in both the co-treated groups. BUN levels remained unaffected following α-tocopherol co-treatment.
Endometritis is one of the leading causes of reproductive inefficiency, infertility and decreased milk yield in high yielding dairy cows and buffaloes as well as in other species of animals and are associate with severe economic loss. Indiscriminate use of antimicrobial agents has resulted in development of resistance to the available antimicrobials. Plant-derived active principles have promising potential to act as an alternative to conventional antimicrobial agents. In the present study, an attempt was made to investigate the mechanism of action of Eucalyptus robusta leaves (ER) extract against resistant bacterial isolates. Out of 70 clinical samples of uterine discharges from cattle and buffaloes having history of uterine infection, we have identified one isolate as methicillin-resistant Staphylococcus. As per the CLSI guideline, this isolate (S4) was found to be catalase +ve, oxidase –ve and possess mecA and coaA genes along with shown resistance to cefoxitin and MIC breakpoint for oxacillin was >4 µg/ml. Methanolic extract of ER reconstituted in water showed presence of eucalyptol (marker compound) along with other major bioactive phytoconstituents following GC-MS analysis. ER extract exhibited marked in vitro antibacterial effect against S4 isolate and MRSA (ATCC 33591) as evidenced by agar well diffusion test with MIC value of 0.3125 mg/ml.
Wound healing is a complex phenomenon with three distinctive yet overlapping phases i.e. Inflammatory, proliferative and remodeling phases. Present study was conducted to unravel the molecular insights of Mangiferin in immunocompromised excisional rat model. Firstly, immunocompromised state was established following hydrocortisone injection @80 mg/kg body weight through intra muscular route and determining the immunocompromised state with neutrophilia, lymphocytopenia and reduced splenic weight in adult male rats weighing 200-250 grams. Then, 120 animals were randomly and equally divided into five groups i.e. group I served as Healthy control, group II served as Immunocompromised control, group III served as Immunocompromised group treated with silver sulphadiazine and group IV and V served as Immunocompromised group treated with 2.5 % Mangiferin and Immunocompromised group treated with 5 % Mangiferin, respectively. Study was conducted for a period of 28 days. Six animals were humanely sacrificed at weekly interval till day 28th of study. Wound healing efficacy and molecular cascading of pharmacodynamics of Mangiferin was determined following gross studies including digital photography, wound area measurement and per cent wound contraction and biochemical indices viz. hydroxyproline, DNA content, total protein and LPO assay.
The present study was undertaken to investigate the antibacterial effect of Eucalyptus robusta leaves extract loaded-solid lipid nanoparticles. Eucalyptus robusta leaves extract loaded SLNs was prepared and characterized. The particle size of SLNs was around 200 nm, zeta potential was 566.93 mV and the entrapping efficiency was 81%. Prepared nanoformulation was spherical in shape. Eucalyptus robusta leaves extract loaded SLNs did not exhibit any promising antibacterial activity against clinical isolates of E. coli and Staphylococcus aureus as determined by Agar well diffusion method. Therefore, further studies were undertaken only on crude extract of Eucalyptus robusta leaves. Experimental endometritis in rats was developed by inoculating the clinical isolate of E. coli (1x107 cfu/ml) in to the uterine horns during diestrus stage followed by cervical ligation. Endometritis in rats was confirmed based on presence of visible pus in the uterus, edematous uterine horns, thinning of endometrial lining and presence of bacterial load in uterine tissue homogenate. Eucalyptus robusta leaves crude extract (25 mg/ml) and Ciprofloxacin (50 mg/ ml) were administered orally found to significantly reduce uterine weight and uterine secretion index. But bacterial load in the uterine tissue homogenate was not significantly reduced by both the treatments. Histopathological lesions in uterus also showed efficient induction of endometritis based on presence of inflammatory cells (neutrophils count) in uterine microscopic sections and their number was markedly reduced after treatment with Eucalyptus robusta leaves crude extract and ciprofloxacin. Compared with the effect of ciprofloxacin, Eucalyptus robusta produced almost similar curative and protective effects against endometritis. Cyclooxygenase 2 (COX-2) gene expression was found to be markedly reduced after treatment with Eucalyptus robusta leaves crude extract as compared with ciprofloxacin treatment. Based on our findings, it may be inferred that Eucalyptus robusta leaves crude extract possesses promising anti-inflammatory activity against experimental endometritis in rats and, therefore, it can be exploited in drug development program for treatment of endometritis in animals.
The objective of the present study was to explore the therapeutic potential of hot alcoholic (100%), hydroalcoholic (70%) and cold aqueous extracts of Pterocarpus marsupium and hot aqueous extract of ITK formulation (mixture of gum acacia, black cumin, wheat and barley) against obese streptozotocin induced type 2 diabetes in male Wistar rats. In Phase I study, tested extracts were observed to possess carbohydrates, alkaloids, proteins and amino acids, tannins, flavonoids, saponins, fixed oils and resins with additional glycosides in ITK formulation after screening by qualitative tests and further confirmed by semiquantitative analysis (by Gas Chromatography-Mass Spectrometry) to possess active principles having antidiabetic, antioxidant, nephroprotective, cardioprotective and anti-inflammatory properties. The extracts were evaluated for in vitro antioxidant (2, 2-Diphenyl-1-picrylhydrazyl (DPPH) and 2, 2’-Azino bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging assay) and antidiabetic (α-amylase and α-glucosidase inhibitory activity) was determined. Aqueous and hydroalcoholic extracts have potent antioxidant and alcoholic extract of P. Marsupium showed superior antidiabetic activity over ITK formulation. Further in Phase II study, 48 obese male Wistar rats were divided into eight groups viz. group I (Healthy Control), group II (Obese), group III (Obese Diabetic), group IV (Obese Diabetic + Metformin @ 50 mg/ kg b.wt.), group V (Obese Diabetic + ITK @ 435 mg/kg b.wt), group VI (Obese Diabetic + Aqueous P.marsupium @ 1 g/kg b.wt) and group VII and VIII were treated with hydroalcoholic and alcoholic @ 300 mg/kg b.wt.
Notch signalling is an evolutionary conserved pathway that allows cell communication through cell-to-cell interactions. Besides its established role in embryonic development, recent reports have shown its involvement in different infections and inflammations of different body systems including cardiovascular system. Sepsis, a systemic inflammatory response syndrome against microbial infection, causes multiple organ failure resulting in death worldwide in human and animals. Vascular hyporeactivity and reduced peripheral resistance is considered to be the leading cause of death in sepsis. Though Notch signalling has been studied especially in macrophages during inflammation including endotoxaemia, its role in vascular beds in sepsis is yet to be explored. Thus, present study was undertaken to evaluate the effect of sepsis on aortic Notch signalling and its involvement in regulating vascular functions during late phage of sepsis. Sepsis was induced by caecal ligation and puncture (CLP) in mice and thoracic aorta was collected for mRNA expression, western blot and immune-histochemical (IHC) study. Functional study on isolated aortic rings from different groups was also performed to assess the vascular reactivity to different vasoconstrictors.
Present study was undertaken to unravel the influence of experimentally induced pre- existing diabetes on survival time and/or mortality pattern in septic animals and how diabetes and sepsis, when occur concurrently, alter vascular reactivity of aorta. Type-1 diabetes was induced following intraperitoneal injection of streptozotocin (STZ: @ 65 mg/kg) for 5 consecutive days while sepsis was induced by caecal ligation and puncture (CLP). Streptozotocin produced a sustained hyperglycaemia in mice and there was decrease in body weight with progression of hyperglycaemic state. CLP caused hyperglycaemia followed by euglycaemia and hypoglycaemia, while in diabeto-sepsis hypoglycaemia was set earlier, leading to early mortality. Alteration in vascular reactivity and endothelial dysfunction was observed.
Present study was undertaken to assess the effect of sepsis on purinergic signaling with special reference to P2Y6 and P2X7 receptors in mouse aorta; and to unravel the interplay between purinergic receptor (P2Y6) and angiotensin II type I receptor (AT1R) in mediating vascular hyporeactivity during sepsis. Sepsis significantly decreased total erythrocytes count, haemoglobin level, total leukocytes count and differential leukocytes count compared to sham-operated mice (SO). Sepsis significantly increased plasma levels of ALT, AST, BUN, creatinine values. Similarly, sepsis significantly increased total bacterial count in peritoneal lavage, blood and spleen compared to healthy control. Functional studies revealed that in mice of SO groups, high K+-depolarising solution produced almost equal contraction in the aortic rings having the intact-endothelium or denuded. But compared to the SO group, mice of the sepsis group, irrespective of the presence or absence of endothelium in aortic rings, exhibited significantly reduced contractile response following exposure to high K+-depolarising solution.
The objective of present study was to evaluate the influence of pre-exposure of chromium @ 1 mg/kg body weight and arsenic @ 38 ppb in drinking water continuously for 30 days in experimentally streptozotocin-induced type 2 diabetes in male Wistar rats and after 30 days of continuous exposure ameliorative potential of an ITK formulation @ 435 mg/kg body weight and its comparative efficacy with metformin @ 50 mg/kg body weight on diabetes induced testicular dysfunction. Eighty four obese male Wistar rats were divided into fourteen groups viz. Normal Control, Obese, Non-diabetic + Chromium, Non-diabetic + Arsenic, Non-diabetic + Chromium-Arsenic combination, Obese Diabetic + chromium, Obese Diabetic + Arsenic, Obese Diabetic + Chromium-Arsenic combination, Obese Diabetic + Chromium + Metformin, Obese Diabetic + Arsenic + Metformin, Obese Diabetic + Chromium-Arsenic combination + Metformin, Obese Diabetic + Chromium + ITK, Obese Diabetic + Arsenic + ITK, Obese Diabetic + Chromium-Arsenic combination + ITK, consisting six animals in each group, were induced diabetes with streptozotocin @ 30 mg/kg body weight, intraperitoneally single dose after 30 days of pre-exposure to chromium and arsenic. After induction of diabetes ITK formulation and metformin were given by oral gavage continuously for 30 days.
The present study was designed to assess the cardio-protective role of oleic acid in myocardial injury. Myocardial injury was induced in rats by intra-peritoneal injection of isoprenaline (ISO; @ 110 mg/kg b.wt) for two consecutive days at 24 h interval. Oleic acid was administered orally (@ 5 mg/kg b.wt or 10 mg/kg b.wt) for 21 days before inducing myocardial injury to evaluate its ameliorative potential. Samples (blood, heart) were collected from different groups of experimental animals 24h after last injection of isoprenaline. Besides evaluation of heart weight to body weight (HW/BW) ratio, myocardial infarct size, oxidative stress parameters and haemato-biochemical parameters, cardio specific biomarkers of injury, ECG, isolated right atrial response and mRNA expression of gene coding for cardiac uncoupling protein-2 (UCP-2) were also quantified. Isoprenaline administration significantly increased the HW/ BW ratio, myocardial infarct size, lipid profiles (total cholesterol, HDL-C, triglyceride) in ISO-induced myocardial injured rats.
The objective of the present study was to explore the therapeutic potential of Tribulus terrestris and ITK formulation against obese streptozotocin- induced type II diabetes and diabetic nephropathy in male Wistar rats. In Phase I study, 70% hot ethanolic extract of Tribulus terrestris fruits and hot aqueous extract of ITK formulation (ingredients gum acacia, black cumin, wheat and barley) was prepared and evaluated for in vitro antioxidant and antidiabetic activity of extracts. Extracts have potent antioxidant (studied by 2, 2-Diphenyl 1-picrylhydrazyl (DPPH) and 2, 2’-Azino-bis (3-ethylbenzothiazoline-6 sulphonic acid) (ABTS) radical scavenging assay), antidiabetic (determined by in vitro α-amylase and α-glucosidase inhibitory activity) and glucose uptake potential in cultured lymphocytes and were observed to possess carbohydrates, alkaloids, proteins and amino acids, tannins, flavonoids, saponins, fixed oils and resins in Tribulus terrestris fruits with additional glycosides in ITK formulation after screening by qualitative tests and further confirmed to possess active principles having antidiabetic, antioxidant, nephroprotective, cardioprotective and anti-inflammatory properties.
The objective of the present study was to explore the therapeutic potential of Gymnema sylvestre and ITK formulation against obese streptozotocin-induced type II diabetes and diabetic cardiomyopathy in male Wistar rats. In Phase I study, 80% hot methanolic extract of Gymnema sylvestre leaves and hot aqueous extract of ITK formulation (ingredients gum acacia, black cumin, wheat and barley) was prepared and evaluated for in vitro antioxidant and antidiabetic activity of extracts. Extracts have potent antioxidant (studied by 2, 2-Diphenyl 1-picrylhydrazyl (DPPH) and 2, 2’-Azino-bis (3-ethylbenzothiazoline-6 sulphonic acid) (ABTS) radical scavenging assay), antidiabetic (determined by in vitro α-amylase and α-glucosidase inhibitory activity) and glucose uptake potential in cultured lymphocytes and were observed to possess alkaloids, f lavonoids, saponins, anthraquinones and phenols in Gymnema sylvestre leaves with additional resins, tanins, proteins, glycosides and fixed oils and fat in ITK formulation after screening by qualitative tests and further confirmed to possess active principles having antidiabetic, antioxidant, cardioprotective, nephroprotective, and anti-inflammatory properties.
The present study was carried out to assess the role of endogenous cannabinoids in regulating aortic vascular response during early and late phases of sepsis. Polymicrobial sepsis was induced by caecal ligation and puncture (CLP) in mice. Besides, recording the isometric tension in arterial rings, estimation of haemato-biochemical parameters, histopathological examinations of vital organs, mRNA expression of CB1 receptor and MAGL enzyme and their role in the vasoconstrictor response to noradrenaline in the aorta of septic mouse were studied. Sepsis significantly reduced RBC, Hb and WBC counts during both early (CLP-6h) and late (CLP-20h) phases of sepsis whereas neutrophil count was increased during early phase. There was also a marked fall in lymphocyte count during late phase of sepsis indicative of immunosuppressive state. Significant rise in the plasma ALT, AST, BUN and creatinine levels during early and late phases of sepsis were suggestive of liver and kidney dysfunctions which were further substantiated byhistopathological examinations of these vital organs. Sepsis produced a state of hypoproteinaemia with significant reduction in plasma albumin level.
Present study was undertaken to evaluate the ameliorative potential of α-tocopherol, curcumin and/or in combination with cisplatin following 28 days continuous exposure. Forty-eight male wistar rats of 190-210 g were divided into eight groups group I (NSS, i.p.), group II (corn oil, oral gavage) served as vehicle control, group III received cisplatin, group IV (α-tocopherol), group V (curcumin), group VI (cisplatin + α-tocopherol), group VII (cisplatin + curcumin), group VIII (cisplatin + curcumin + α-tocopherol). Cisplatin was given by intra-peritoneal route @ 0.5mg/kg b.wt., while α-tocopherol @ 100 mg/kg b.wt. and curcumin @ 50 mg/kg b.wt. by oral gavage continuously for 28 days. Cisplatin produced apparent signs of toxicity like rough coat, cachexia, decreased activity but no mortality in rats. Body weight and percent weight gain in rats with cisplatin alone and in combination with α-tocopherol and curcumin treated groups were significantly lower. Absolute and relative organ weight did not differ between different groups. Significant reduction in Hb on 21 day and 28 days after exposure was observed. Marked reduction in PCV, TLC and platelet count in cisplatin alone treated group following 28-day exposure however DLC remain unaltered. Significant decrease in creatinine clearance in all cisplatin exposed groups when compared to control groups.
Present study was embarked upon to extricate the underlying calcium signaling mechanisms responsible for spasmogens (PGF2α , and oxytocin) induced muscular contractions in oviducts of non-pregnant buffalo in oestrous stage. Isometric tension in longitudinal oviductal in both ampulla and isthmus strips was recorded under the resting tension of 1 ± 0.2 g following mounting the tissue in Ringer–locke solution. Following an equilibration period of about 120 to 150 min, oviductal strips both ampulla and isthmus presented a consistent and rhythmic pattern of spontaneity. Mean Integral Tension (MIT) and amplitude (g) of ampulla and isthmus in normal spontaneous tissue did not differ significantly. The frequency (BPM) of ampulla and isthmus in normal spontaneous tissue differed significantly (P<0.05). PG2α produced significantly (P<0.05) greater contraction than oxytocin in both ampulla and isthmus of buffalo oviduct in concentration-dependent manner. In isthmus the contraction produced by oxytocin was negligible. PGF2α produced a significant change in tonic contraction between ampulla and isthmus but the change in phasic contraction between ampulla and isthmus was not significant. PGF2α produced significantly (P<0.05) greater contraction in ampulla than in the isthmus of buffalo oviduct.
Hydrogen sulphide, a gas with rotten egg smell, is traditionally considered as a toxicant and an environmental pollutant but recently, it has gained attention as a mediator of physiological and biological processes. The present study was undertaken to unravel the effect of H2 S on myometrial activity and its underlining mechanism in non-pregnant buffaloes. L-cysteine is considered to be main endogenous amino acid responsible for hydrogen sulphide formation due to the action of two cytosolic enzymes- cystathione ? synthase (CBS) and cystathione y lyase (CSE/CGL). Sodium hydrogen sulphide and GYY4137 are major hydrogen sulphide donors were used in this present study. In vitro- exposure of isolated buffalo myometrial strips to L-cysteine (10nm to 3mM) produced concentration- dependent uterotonic effect. L-cysteine induced uterotonic action is dependent on the activity of cystathione ? synthase (CBS) and cystathione y lyase (CGL/CSE) as evidenced by rightward shift of the DRC of L-cysteine in the presence of enzyme blockers (AOAA and PAG, respectively) it is to be further noted that following blockade of CBS and CSE enzyme, possibly some relaxant mechanism may get activated by L-Cysteine to produce uterine relaxation. The existence of CBS enzyme of molecular weight, 63kDa and CSE/CGL of 45kDa was observed by western blot technique. L-cysteine failed to produce any appreciable contraction in the absence of extracellular calcium. Further, in the presence of nifedipine, the uterotonic action of L-cysteine was completely abolished.
Present study was undertaken to investigate the modulatory effect of Eucalyptus citriodora leaves extract and Cefixime on inflammatory biomarkers in experimentally induced endometritis in Wistar rats. Uterine discharge of clinical and subclinical endometritis cases having history of repeat breeding (43%) and abortion (57%) were found to have E. coli (60%) Staphyloccocus aureus (40%). Eucalyptus citriodora leaves methanolic extracts exhibited promising antibacterial activity against both clinical isolates as determined by disc diffusion method. Rat endometritic model was developed by inoculating the mixture of E. coli (1? 106)and Staphloccocus aureus (1? 108)in to uterine horns during diestrus stage followed by cervical ligation and the model was confirmed based on presence of visible pus in the uterus, edematous uterine horn, thinning of endometrial lining and presence of large number of polymorphonuclear cells and bacterial load in uterine flushing.
