Veterinary Research Forum

Abstract Micromeria congesta (MC) is a medicinal plant traditionally used in the Şanlıurfa, Adıyaman, and Gaziantep provinces of Türkiye for treating respiratory disorders. Despite its ethnobotanical relevance, its pharmacological effects on lung inflammation remain unclear. This study evaluated the protective effects of MC essential oil (MCEO) against lipopolysaccharide (LPS)-induced acute lung injury in rats, with a focus on inflammation and tissue damage. The MC was collected from its natural habitat, and its EO was extracted by hydro-distillation using a Clevenger-type apparatus. The Gas chromatography-mass spectrometry analysis revealed 28 compounds, with piperitenone oxide (33.97%) and pulegone (22.30%) as the main constituents. Thirty-six Wistar albino rats were randomly assigned into six groups, including three MCEO-treated (12.50, 25.00, and 50.00 mg kg^-1), dexamethasone, LPS-only, and control groups. The MCEO was administered by gavage for seven days, followed by intra-venous LPS on day eight (except for the control group). Blood and lung tissues were collected 6 hr later for biochemical, histopathological, and molecular analyses. The MCEO treatment significantly lowered blood glucose and urea levels compared to the LPS group. It also modulated NLRP3, caspase-3, and tumor necrosis factor alpha genes expressions, as determined by RT-PCR analysis, indicating anti-inflammatory effects. Histopathological findings confirmed a dose-dependent reduction in lung inflammation and tissue damage. The MCEO administration alleviated LPS-induced acute lung injury in rats through anti-inflammatory and protective effects, as confirmed by molecular (NLRP3, caspase-3, and tumor necrosis factor alpha) and histopathological analyses. Additionally, MCEO reduced blood glucose and urea levels, supporting its systemic protective role against endotoxin-induced inflammation.

Abstract The administration of dispersed allogeneic biomaterial (AB) into the para-articular region and joint cavity allows slowing down the processes of tissue destruction in arthritis. The aim of the study was to examine the effect of AB on the course of experimental adjuvant arthritis (AA) in rats. For modeling of AA, complete Freund's adjuvant was injected into the plantar surface of the hind paw of 60 white outbred female rats. The study included intact group, control group, and experimental group. After 37 days of the experiment, blood was collected for hematological analysis and the knee joint with surrounding tissues was harvested for standard histological examination. Intra-articular administration of AB to experimental rats while using complete Freund's adjuvant neutralized the manifestation of signs of a generalized inflammatory process in the joints and reduced the degree of destructive changes in the articular apparatus, preserving the structure of the cartilaginous layer. The use of AB made it possible to stabilize the red and white blood cells levels in the experimental group, as well as significantly increase the reduced level of monocytes. Intra-articular administration of AB during AA modeling exhibits an osteo- and chondro-protective effect, providing positive anti-inflammatory and symptom-modifying effects and weakening the manifestation of pathomorphological changes in the joints of experimental rats.

Abstract Intense exercise is known to induce oxidative stress and inflammation, potentially leading to liver damage. This study examined the potential protective effects of apilarnil (AP), a natural bioactive compound with antioxidant and anti-inflammatory properties, against exercise-induced liver injury. Thirty-five male Wistar albino rats were allocated into five groups: Control, non-exercise (NEX), exercise (EX), EX + AP1 (0.20 g kg^-1) and EX + AP2 (0.40 g kg^-1). At the end of the 14-day experiment, serum and liver tissue samples were collected for the analysis of histopathological changes, oxidative stress markers, inflammatory cytokines and serum biochemical parameters. Histopathological evaluations revealed substantial liver damage in the EX group. However, in the EX + AP1 and EX + AP2 groups, the severity of these lesions was significantly attenuated. Biochemical analyses demonstrated elevated levels of tumor necrosis factor-alpha, interleukin-1 beta and interleukin-6 in the NEX group were markedly reduced by AP supplementation. Similarly, malondialdehyde levels were increased, while the activities of antioxidant enzymes - catalase, superoxide dismutase and glutathione peroxidase were declined in the NEX group. AP supplementation reversed these effects by lowering malondialdehyde levels and enhancing antioxidant enzyme activities in the EX + AP1 and EX + AP2 groups. Additionally, serum biochemical analyses indicated improved lipid profiles and liver function parameters in the AP -treated groups compared to the NEX group. In conclusion, histopathological and biochemical findings indicated that AP supplementation mitigated exercise-induced liver damage by reducing oxidative stress and inflammation, while enhancing antioxidant defenses.

Abstract Diazinon (DZN) is a widely used organophosphate. We studied the effect of quercetin pegylated liposome (QPEGL) on acute low dose DZN-induced oxidative stress and behavioral disorders through monitoring brain serotonin metabolite 5-hydroxyindoleacetic acid (5-HIAA) in mature male rats. Animals were treated in two control groups that received a single dose of normal saline and dimethyl sulfoxide, and four groups that received a single dose of DZN 10.00 mg kg^-1 (DZN), DZN 10.00 mg kg + quercetin 20.00 mg kg^-1, DZN 10.00 mg kg^-1 + PEGL 20.00 mg kg^-1, DZN 10.00 mg kg^-1 + QPEGL 20.00 mg kg^-1 (QPEGL), respectively. Performances of the rats were investigated by the open field and elevated plus maze tests. Twenty-four hr after the treatments, animals’ brains were harvested and frozen at – 80.00 ˚C. Brain tissues 5-HIAA level was determined by the enzyme-linked immunosorbent assay. Furthermore, malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GPx) levels were determined for oxidative stress analysis. The motor activity was significantly reduced in the DZN group compared to the control group following increased anxiety-like behavior and ameliorated by QPEGL. Moreover, 5-HIAA and MDA levels notably increased in the DZN group compared to the control group and significantly decreased in the QPEGL group compared to the DZN group. The SOD and GPx contents were not significantly changed in the DZN group compared to the control; although, these parameters improved after treatment with QPEGL. Acute low dose DZN exposure resulted in lipid peroxidation and elevated levels of the serotonin metabolite (5-HIAA), leading to neurobehavioral disorders, such as anxiety-like behavior and impaired motor activity, which were alleviated by QPEGL.

Abstract Tarantula cubensis alcoholic extract (TCAE) is a homeopathic product used in the veterinary field. This study aimed to determine the effects of TCAE on cadmium (Cd) toxicity in the embryo. The study used 220 fertile, incubated chicken eggs divided into 11 equal groups on the 7^th day of incubation. The groups comprised untreated and physiological saline control groups, a group with TCAE alone, four groups with varying doses of Cd alone and four groups with the same doses of Cd plus TCAE. At the end of the incubation period, the eggs were opened, kidney and liver tissue samples were taken for histopathology and the number of dead and living embryos were recorded. In the prsent study, the median lethal dose of Cd was determined to be 0.029 mg per egg and the median lethal dose of Cd plus TCAE was determined to be 0.020 mg per egg. The histopathological examinations determined that kidney and liver damage were increased when TCAE and Cd were administered together, that was higher than when Cd was given alone. Thus, TCAE, which had no toxic effect on the embryo when used alone, might increase the embryotoxic activity of Cd. However, more detailed studies are needed.

Abstract Phthalate esters, such as dibutyl phthalate (DBP), are extensively utilized and human and animal exposure leads to serious toxic effects, including hepatotoxicity. In the present study the protective effects of royal jelly (RJ) on DBP-induced liver damage was investigated. A total number of 40 Wistar albino rats were randomly divided into eight groups (n = 5): control (corn oil), DBP (500 mg kg^-1), RJ (200 mg kg¹), Quercetin (QCN; 50.00 mg kg^-1), RJ (100 mg kg^-1) + DBP, RJ (200 mg kg^-1) + DBP, RJ (300 mg kg^-1) + DBP, QCN (50.00 mg kg^-1) + DBP. After 28 days of daily oral gavage treatment, animals were euthanized. The insulin resistance index, lipid profile and hepatic enzymes were measured on the collected serum samples. Moreover, oxidative and nitrosative stress biomarkers were determined in the liver. Histopathological alterations and ultimately cytochrome P450 2E1 (CYP2E1) activity was also assessed. Data obtained revealed that RJ significantly reduced the insulin resistance index and liver enzymes level in RJ-DBP groups. At the same time, RJ recovered the DBP-induced oxidative stress and restored the DBP-depleted glutathione. Moreover, RJ improved lipid profile and reduced significantly the DBP-induced hepatic CYP 2E1 activity in RJ-DBP groups. Dibutyl phthalate induced-hepatic damage such as necrosis of hepatocytes and scattered bleeding was alleviated in RJ-DBP group. Our data suggested that the administration of RJ could protect the DBP-induced hepatic functional and structural alterations. The RJ protective effects might be attributed to its antioxidant and anti-inflammatory properties and reduced CYP 2E1 activity.

Abstract Pulmonary fibrosis (PF) is a chronic interstitial lung disease with a progressive damage to the air sacs and deposition of collagen fibers in the lung tissue. The study aimed to explore the effects of Nigella sativa oil (NSO) or thymoquinone (TQ), alone or in combination with dexamethasone (DEX), on the development of bleomycin (BLM)-induced PF. Forty-two male rats were divided into seven groups: Control (CTRL); BLM, received a single dose of BLM on day 0, intratracheally; all remaining groups received BLM, as well. DEX, received DEX daily, intraperitoneally, 1 day before BLM and continued for 14 days; NSO and TQ groups, received daily NSO and TQ, respectively, 7 days before BLM and continued for 35 days; DEX + TQ, received both DEX and TQ; DEX + NSO, received both DEX and NSO. At the end, lung tissues were used for histopathological and biochemical analyses. BLM significantly increased the severity of fibrosis and inflammation compared to the CTRL. Bleomycin also significantly increased the amount of hydroxyproline, however, decreased most antioxidant enzymes in the lung tissue compared to the other groups. Group TQ + DEX significantly reduced the severity of BLM-induced PF as well as alterations in biochemical parameters, lung weight and O2 saturation. Nigella sativa oil slightly reduced BLM-induced PF, however, it caused non-significant hyperemia in lung tissue. Thymoquinone potentiated the effects of DEX on most biochemical and pathological alterations of BLM-induced lung injury much better than NSO. More studies are needed to support the use of NSO and TQ as potential protective agents against PF.

Abstract Docetaxel (DTX) is widely utilized in breast cancer treatment. However, cancer cell resistance has limited its anti-tumor efficacy. Some molecules called microRNAs (miRNAs), acting like fine-tuned switches, can influence how breast cancer develops and spreads. We conducted a study to examine if augmenting breast cancer cells with a particular molecule, known as miRNA-142-3p, could improve the efficacy of a widely used treatment called DTX. The expression level of miR-142-3p was initially assessed in MDA-MB-468 cells. The miRNA transfection was performed to conduct additional experiments. The impact of a combined treatment involving DTX and miRNA-142-3p on both cell migration (by wound healing assay) and apoptosis (using annexin V/Propidium iodide staining) was examined. Cell viability was determined through the MTT assay, and gene expression was quantified using quantitative real-time polymerase chain reaction. The combined application of DTX and miRNA-142-3p resulted in a significant decrease in the expression of factors promoting tumor growth, such as SOX2, Octamer 4, HMGA2, Kruppel-like factor 4, and Bach-1. Additionally, the combination of miRNA-142-3p and DTX initiated apoptotic cell death. Moreover, the progression of breast cancer cells was impeded by inducing cell cycle arrest at the G¹ phase. This combination also efficiently restrained the migration and invasion of breast cancer cells. The DTX or miRNA-142-3p alone can suppress malignant behavior and progression of breast cancer cells, but their combination elicits a synergistic effect that further enhances breast cancer inhibition. In summary, miRNA-142-3p transfection can be administered in conjunction with DTX therapy to enhance its cytotoxicity against breast cancer cells and prevent chemoresistance.

Abstract In this study, a controlled released delivery drug system designed and synthesized by loading curcumin and selenium nanoparticles (SeNaPs) on chitosan hydrogel, and while evaluating the physicochemical properties of the prepared drug delivery system, the tissue changes caused by the local implant of that system in rats with experimental spinal cord injury (SCI) were investigated. For this purpose, 100 adult female rats were randomly divided into five equal groups which are: Control group without any treatment for SCI, chitosan group that received chitosan hydrogel, curcumin group that received curcumin-loaded hydrogel, SeNaP group that received chitosan loaded with SeNaPs and SeNPCur group that received chitosan loaded with SeNaPs and curcumin. On the 3^rd and 7^th days of the study, severe infiltration of leukocytes, especially lymphocytes, as well as axon swelling and hemorrhagic necrosis at the lesion sites were observed in all groups, especially the control group. On the 7^th day, the severity of these injuries decreased in the SeNPCur group and the highest number of astrocytes was observed in this group. In addition, on the 14^th and 21^st days of the study, the lowest severity of nerve tissue damage and the lowest presence of inflammatory cells along with the highest number of astrocytes were seen in the SeNPCur group. The glial fibrillary acidic protein study also confirmed the presence of more and significant astrocytes in the SeNPCur, curcumin and SeNP groups at different times of the study, respectively. The histopathological results showed the neuroprotective effects of chitosan hydrogel loaded with selenium and curcumin.

Abstract Aminoglycoside antibiotics (AGs) can cause neuromuscular blockade and paralysis of skeletal muscles. To compare the paralytic effects of selected AGs on some motor behaviors in mice, 24 male mice weighing 20.00 to 25.00 g were divided into four treatment groups. Each group was given one of four AGs (gentamicin, dihydrostreptomycin, apramycin and amikacin) at incremental doses that increased half-logarithmically compared to the therapeutic dose (16.00 mg kg^-1). Motor behavioral tests included open field test, inclined plane, horizontal bars, static rods, parallel bars and rotarod. Finally, the data were analyzed using descriptive and analytical statistics. Gentamicin and dihydrostreptomycin at 32.00 times of the therapeutic dose produced complete paralysis of the limbs, respiratory arrest, and even death in some animals. However, apramycin and amikacin did not show significant effects on skeletal muscle and motor behaviors at 32.00 times of the therapeutic dose. After administration of apramycin at 100 times of the therapeutic dose, four out of six mice (66.67%) died from respiratory depression. Amikacin at this dose did not cause animal death, although it caused some changes in motor behaviors with a significant difference in comparison with control values. Gentamicin demonstrated significantly more potent effects on motor behaviors compared to the other AGs. Overall, the order of potency was gentamicin > dihydrostreptomycin > apramycin > amikacin. High doses of AGs could impair the skeletal muscle function and disrupt motor behaviors in mice. Furthermore, the paralytic potency of selected AGs on skeletal muscle was significantly different.

Abstract The present study investigated the prospect of improvement in pharmacokinetic (PK) parameters of marbofloxacin due to alpha-1-monolaurin pre-treatment in broiler chickens. Two groups of broilers were administered a single oral dose of marbofloxacin (5.00 mg kg^-1 body weight): Group-I without pre-treatment and Group-II with alpha-1-monolaurin pre-treatment (4.00 g kg^-1 feed for 10 days). Blood sampling was done periodically for both groups and plasma marbofloxacin concentrations were determined using ultra-high performance liquid chromatography. Pharmacokinetic parameters using non-compartmental modelling approach were calculated with the PKSolver software. Statistical analysis revealed significant differences in plasma marbofloxacin concentrations between the two groups at 1, 2, and 24 hr. Group-II birds exhibited a higher mean maximum plasma concentration (2.43 µg mL^-1) at an earlier time (T_max: 1.38 hr) compared to Group-I. The plasma concentrations of marbofloxacin were maintained above 0.10 and 0.18 µg mL^-1 up to 24 hr in Group-I and Group-II broilers, respectively. Significant differences were observed in PK parameters such as the area under the curve and total body clearance. The mean relative oral bioavailability of Group-II birds compared to Group-I was 119.61%. The findings of the study provided evidence of PK parameters enhancement of marbofloxacin in the alpha-1-monolaurin pre-treated group. The calculated PK-pharmacodynamic indices for marbofloxacin predicted clinical efficaciousness in the broiler chickens.

Abstract In the present study the protective effect of Nigella sativa (N. sativa)on synaptic plasticity impairment induced by lipopolysaccharide (LPS) in rats was investigated. Fifty-eight rats were grouped and treated as follows: 1) control (saline), 2) LPS, 3) LPS-N. sativa,and 4) N. sativa. In a Morris water maze test, the escape latency and traveled path to find the platform as well as time spent and the traveled distance in target quadrant (Q₁) were measured. Long term potentiation (LTP) from CA₁ area of hippocampus followed by high frequency stimulation to Schafer collateral was studied and slope, slope 10-90% and amplitude of field excitatory field potential (fEPSP) were calculated. The escape latency and traveled path in LPS group were significantly higher than those in the control group while, in LPS-N. sativa group these parameters were significantly lower than those in LPS group. The rats in LPS group spent less time and traveled shorter distance in Q₁ than the rats in the control group while, in LPS-N. sativa group the rats spent more time and traveled longer distance than the rats in LPS group. LPS significantly decreased slope, slope 10-90% and amplitude of fEPSP while, in LPS-N. sativa group these parameters increased compared to LPS group. The results indicated that the hydro-alcohol extract of N. sativa protected against synaptic plasticity and spatial learning and memory impairment induced by LPS in rats.

Abstract Busulfan is an alkylating agent affects ovarian follicles growth by oxidative stress induction. Satureja khuzestanica has antioxidant effects. The aim of this study was to examine whether S. khuzestanica essential oil (SKEO) exhibits protective effects on busulfan-induced ovarian failure. Eighty-four adult female mice were divided into six groups including dimethyl sulfoxide (control), SKEO 225.00 mg kg^-1 (orally), busulfan 3.00 mg kg^-1 (orally), busulfan 36.00 mg kg^-1 (intraperitoneally), busulfan 3.00 mg kg^-1 and SKEO and busulfan 36.00 mg kg^-1 and SKEO. After 28 days, the mice were euthanized and oocytes were removed for in vitro fertilization (IVF) rate evaluation. Oocyte quantity and quality, fertilization rate and pre-implantation embryo development were daily examined with a stereo microscope in a period of 120 hr. Serum levels of estradiol and progesterone were also evaluated. Busulfan caused significant decreases in oocyte number and quality, fertilization rate, pre-implantation embryo development and embryo quality. The SKEO significantly decreased the adverse effects of busulfan. The present study indicated that SKEO can protect female fertility potential against busulfan induced damages.

Abstract Bisphenol-S (BPS) is a new bisphenol-A substitute widely used in many plastic products. Bisphenol-A as a main member of bisphenol family has been known as an endocrine system disrupter chemical compound. Like other members of bisphenol family, there is public health concern about the toxic effects of BPS on reproductive system, thus, we examined BPS effects on in vitro fertilization (IVF) potential and oxidative stress status in a murine model. Adult female mice (n = 70) were randomly divided into control and BPS-treated groups. Bisphenol-S was administered at doses of 0, 1, 5, 10, 50 and 100 µg kg^-1 body weight per day intraperitoneally for 21 consecutive days. Twenty-Four hr after the last treatment, five mice in each group were super-ovulated and the oocytes were harvested for IVF. All ovaries were collected and used for biochemical factors analyses. Bisphenol-S exposure at doses more than 10 µg kg^-1 induced developmental arrest of pre-implantation embryos. Further, lipid peroxidation measurement in ovaries indicated that all doses of BPS cause oxidative stress in female mice. In conclusion, BPS administration even in low doses can result in female reproductive toxicities and oxidative stress in mice.

Abstract Application of smoke condensate derived from an indirect heating of jennet feces (Sargin) had been recommended by Iranian ancient scientists as a therapeutic agent. The present study was done to evaluate the healing effects of Guajol^® ointment on burn wound in rat. The Guajol^® ointment was prepared from the smoke condensate of Sargin samples. Wistar Rats (n = 50) were randomized into six groups including normal saline, silver sulfadiazine and 1.25%, 2.50%, 5.00% and 10.00% concentrations of Guajol^® ointment. Under general anesthesia, dorsum of the rats were shaved and burn wounds were created using hot plate. Area of wounds and percent of healing were measured. Normal saline had the highest area of wound, followed by 1.25% Guajol^® and silver-sulfadiazine groups. The group treated with 5.00% Guajol^® showed the highest percent of healing. Percent of healing in NS, SSD and 5.00% Guajol^® ointment groups on day 21 were 38.47%, 75.00% and 98.51%, respectively. Microscopic examination of wounds sections of rats treated with 5.00% Guajol^® showed more collagen fibers and fibroblasts cells on day 7. Wounds of 5.00% Guajol^® treated group was covered with healthy epithelial and epidermis tissues and hair follicles on day 21. This was the first report of using Sargin to heal the burn wound of rat. Further studies are recommended for investigation of the other effects of Guajol^® ointment and its possible application in medicine.

Abstract It has been shown that chronic exposure to cypermethrin (CPM), a pyrethroid pesticide, results in follicular atresia via pathologically affecting angiogenesis, disrupting endocrine potential and enhancing oxidative stress. This study was aimed to uncover the CPM-exposed energy dependent follicular cells apoptosis and to estimate protective effect of vitamin E (VitE) as a potent antioxidant. Thirty six Wistar rats were divided into six groups (n = 6 rats for each group) including; control-sham, CPM-received (CPM, 75 mg kg^-1, intraperitoneally), and CPM and VitE-treated (VitE, 150 mg kg^-1, orally) for 14 and 24 days. The protein biosynthesis of glucose transporter-1 (GLUT-1) and caspase-3 in follicles were estimated by using immuno-histochemical staining at preantral and antral stages. Moreover, the periodic acid Schiff (PAS) staining was performed in order to evaluate the intracytoplasmic carbohydrate ratio in follicular cells and oocyte. Percentages of follicles with GLUT-1, Caspase-3 and PAS-positive cells were compared between groups. Immunohistochemical analyses showed that, VitE significantly up-regulated the GLUT-1 expression and improved the intracytoplasmic carbohydrate supplementation especially at preantral follicles. The cross sections from the CPM-exposed ovaries represented remarkable elevation in percentage of atretic preantral and antral follicles with caspase-3 biosynthesis, which was remarkably (p < 0.05) diminished in VitE co-treated groups. In conclusion, our data showed that VitE by up-regulating of the GLUT-1 biosynthesis improved glucose uptake at follicular cells and oocyte levels that in turn inhibited pro-apoptotic protein caspase-3 biosynthesis.

Abstract In the present study, the effects of intracerebroventricular (ICV) administration of normal saline (control), histamine, mepyramine (a histamine H1-receptor antagonist) and ranitidine (a histamine H2-receptor antagonist) were investigated on the formalin-induced pain in rabbits. Subcutaneous (SC) injection of a formalin (100 μl, 5%) solution into the ventral surface of the right hind paw was performed, and the time durations spent licking and biting the injected paw were measured in 10 min blocks for 1 h. The SC injection of formalin produced a short-lasting (10 min) pain response. The ICV injection of histamine at doses of 25, 50 and 100 μg significantly (P < 0.05) decreased the time duration spent licking and biting the injected paw. Mepyramine and ranitidine, used alone produced no effects. The ICV pretreatments with mepyramine and ranitidine at the same dose of 200 μg significantly (P < 0.05) prevented histamine (100 μg, ICV)-induced antinociception. These results indicate that activation of brain histamine with ICV injection of exogenous histamine produces antinociception. Central histamine H1 and H2 receptors may be involved in the centrally administered histamine-induced antinociception in the formalin-induced pain in rabbits.