فصلنامه آسیب شناسی درمانگاهی دامپزشکی
سال چهاردهم شماره 1 (پیاپی 53، بهار 1399)

The liver performs various pivotal functions, including protein synthesis, glucose homeostasis, detoxification, and the utilization of various nutrients. Generally, when the liver is exposed to high levels of environmental toxins, metabolic dysfunction of the liver may occur, which ranges from the transient elevation of liver enzymes to life‐threatening hepatic fibrosis, liver cirrhosis, and even hepatocellular carcinoma. Substantial evidence implicates oxidative stress and inflammation in the aetiology of liver injury. Similar effects are caused by carbon tetrachloride (CCl4), an industrial solvent known to induce liver injury and liver diseases, which is widely used in experimental hepatopathy. CCl4‐induced toxicity depends on the dose and duration of exposure. At a low dose, transient effects occur, including the loss of Ca2+ sequestration, impaired lipid homeostasis, and the release of several cytokines. Longer exposures alter fatty acid metabolism and induce fibrosis, cirrhosis, and cancer. CCl4‐induced hepatotoxicity is the result of reductive dehalogenation reactions catalyzed by the hepatic cytochrome P‐450, which forms unstable trichloromethyl and trichloromethyl peroxyl radicals capable of binding to proteins or lipids and initiating lipid peroxidation and liver damage. Oxidative stress has been accepted as one of the principal causes of CCl4‐induced hepatic injury, which is mediated by the production of free radical derivatives of CCl4 and is responsible for cell membrane damage and the subsequent release of the marker enzymes of hepatotoxicity. Inflammation is another important pathological mechanism through which CCl4‐induced liver injury is propagated. Plants have a long history of medicinal applications across cultures, and many modern pharmaceuticals were originally derived from botanicals. Fenugreek (Trigonella foenum graecum L.) is an annual legume crop widely cultivated in Asia, Africa and the Mediterranean countries where it is often used for the edible and medicinal values of its seeds. In other countries like Canada and England, fenugreek is used for incorporation into short-term crop rotation due to its high adaptability to dry climatic conditions, annual nature, and ability to fix atmospheric nitrogen in the soil. As a feed for livestock, it is regarded as a bloat-free crop that contains growth-promoting compounds such as steroids and diosgenin, which are not present in other forage legumes. Fenugreek is a plant whose seeds have been historically used in cooking and as a herbal remedy, with anti-microbial, anti-inflammatory, anti-oxidant, anti-cancer, and anti-diabetic effects. Current research on fenugreek, most carried for therapeutic purposes, has shown that it contains beneficial chemical constituents including steroidal sapogenins, fibre, galactomannans, antioxidants, and amino acids such as 4-hydroxy isoleucine which possess antidiabetic, antioxidative, hypocholesterolemic, hypoglycemic, anti-inflammatory, antiulcerogenic, antitumor and immunomodulatory properties. To evaluate fenugreek as a hepatoprotective agent, we investigated the protective effect of fenugreek seed extract against CCl4-induced hepatic toxicity in rats.

A total of 48 male Wistar rats were randomly divided into eight groups. The normal control group was intact. The liver-injured group was received CCl4 (1 ml/kg bw, 50%, i.p.) twice a week for 10 times. Normal experimental groups received fenugreek extract (50, 100 and 200 mg/kg bw, intragastrically). Liver-injured experimental groups administrated fenugreek extract (3 doses as above) along with CCl4. The rats were sacrificed on the 34th day, and blood was withdrawn by cardiac puncture. At the end of the 34-day treatment, the rats were fasted for 12 h and killed under mild (diethyl ether) anaesthesia. Blood was immediately obtained after the animals were sacrificed. The blood was withdrawn by cardiac puncture, allowed to clot and the serum was separated by centrifuging. The level of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and gamaglutamyltransferase (GGT) were measured. Data were analyzed using one-way ANOVA and Tukey test. The criterion was significant .

The vital properties of garlic have been known since ancient times. It is an herbal medicine with antioxidant, anti-hyperlipidemic and antidiabetic properties which is extensively used in the treatment of diseases. Garlic is rich in sulfur compounds, mainly in the form of cysteine derivatives. Among domestic animals, dogs and cats are more sensitive to the oxidative effects of Allium sativum family plants including garlic and onion due to differences in antioxidant defense system of red blood cells (Kovalkovicova et al., 2009). Herbal remedies are usually cheaper than chemical medicines. However, some reports suggest that poisoning and hematological complications may occur following the consumption of raw garlic or its extract in dogs (Lee et al., 2000). The purpose of the present survey was to assess probable hematologic alterations and the activity of some erythrocyte antioxidant enzymes following garlic administration in dogs.

The present study was performed in Veterinary Hospital of Shahid Chamran University of Ahvaz during October-March 2015. For this purpose ten mixed breed dogs were selected with an average age of 1 to 2 years old in a similar weight range (19.6 kg). All the studied dogs were clinically healthy. They were selected from the population of rural dogs around Ahvaz district. Age of dogs was estimated according to dental formulary and tooth wear. In this research, all dogs were treated with garlic tablet (100 mg/kg, only once daily and for 45 days). Blood samples were collected three times on days zero, 45 and 60 of challenge. Complete blood count (CBC), erythrocyte morphology, reticulocyte count, Heinz body and erythrocyte osmotic fragility test were measured in different times. Evaluation of antioxidant enzymes activity of red blood cells including superoxide dismutase (SOD) and glutathione peroxidase (GPX) were performed by commercial kits (Randox, UK) and spectrophotometric method (Optima 3000 plus, Japan). According to the kit instruction, 0.5 ml of whole blood was slipped after three steps of washing with normal saline (NaCl 0.9%) (Merck, Germany), by adding 2 ml of cold distilled water (4° C). The obtained samples were stored at -70° C until testing.

It was determined that the use of garlic tablet with dosage of 100 mg/kg was tolerated in the studied dogs. No clinical complications were observed in animals (such as diarrhea, vomiting and anorexia). Garlic administration caused a significant reduction in erythrocyte count, hematocrit and hemoglobin concentration on day 60 compared to zero day (p

Pica is one of the disorders that has high potential for inducing various damages such as poisoning, digestive accumulation, botulism, weakening of the immune system, weight loss, etc. in ruminants farms and for years, the researcher has encountered this problem in lambs. The present study was conducted to compare the effect of Microvit supplementation and Ferrodrop oral drops on the primary parameters of homogram and serum minerals in 20-days-old healthy and with Pica lams. This clinical trial was performed on 10-headed 20-days-old lambs mixed-breed without Pica and 10-headed 20-day-old lambs mixed-breed with Pica. Both groups received 2 types of oral therapy including Microvit supplementation (7 days, 0/5 g / l of consuming water) and oral furuderap drops (4 days, 300 mg/day in consuming milk). Before starting treatment and after each treatment, blood and serum samples were taken from lambs and sent to the laboratory for determination of CBC and serum mineral elements. Data were analyzed by SPSS statistical software using repeated measures ANOVA at 95% confidence level. In patients with Pica blood Hb, MCV and MCHC and serum iron was less than nonpatients (p0/05), and Pica was not treated in patients (p>0/05). Treatment with furuderap increased the blood Hb, MCV and MCHC and serum iron levels, significantly and Pica was treated in patients (p

Cadmium is a trace metal that chronic exposure to it leads to poisoning in humans and animals. This heavy and toxic metal has deleterious effects on vital organs such as the liver, kidneys and immune system. The purpose of the study was to evaluate the effects of extract and powder of peel of pomegranate on the lipid profile and tissue structure of the kidneys following the experimental cadmium poisoning in Japanese quail. For this purpose, five hundred and ten 7-day-old quail chicks were divided into 10 equal groups. Group 1) and Group 2) were selected as negative control and cadmium control (20 ppm) respectively; Group (3) and Group (4) received 0.1% and 0.2% pomegranate peel extract with 20 ppm of cadmium, Group (5) received 1% pomegranate peel powder with 20 ppm of cadmium; group (6) received 2% pomegranate peel powder along with 20 ppm of cadmium; Group (7) received 0.2% pomegranate peel extract; Group (8) received 0.1% pomegranate peel extract; Group (9) received 1% pomegranate peel powder; Group (10) received 2% pomegranate peel powder. On day 42, blood samples were collected from the brachial vein, and centrifuged at 1500 rpm for 15 minutes. The plasma was isolated from samples and kept at -20°C until lipid profile analysis. The concentration of triglyceride, total cholesterol and high density lipoprotein (HDL) in the serum of birds in different groups were determined using specific kits (Pars Azmoon -Iran). The low density lipoprotein (LDL) was evaluated using Friedewald equation. Then, the birds were slaughtered and the kidney samples were removed from the abdominal area and transferred to formalin. 5 micrometers sections were prepared and stereological study was performed using a point grid. The statistical analysis was performed using One-way ANOVA followed by Tukey multiple comparison tests using 5% level of significance. All results are expressed as mean ± standard deviation (S.D). The results showed that the highest number of proximal tubes was in the 1% pomegranate peel powder group which was significantly higher than the positive control group (p

Infectious by salmonella genus bacteria causes acute and chronic diseases in poultry, and finally causes considerable economic losses to poultry industry. Poultry is one of the important hosts of the salmonella bacteria and most developed countries, is a major cause of contaminated foodborne illnesses. Non-typhoid Salmonellae have been recognized as a major cause of intestinal infections of human origin, a major source of these infections being animal products, especially poultry products (Swayne et al., 2013). The virulence spvRABCD plasmid has been found in Salmonella gallinarum and Salmonella pullorum and is also essential for the development of clinical disease (Rychlik et al., 1998). However, the results of some studies have shown that the role of virulence plasmids in the pathogenesis of salmonella is not clear. But there is evidence that spv genes can infect Salmonella typhimurium in the spleen and liver and increase the rate of bacterial proliferation in host cells (Gulig and Doyle, 1993). The aim of current study was to evaluate antibiotic resistance in infected layer flocks by salmonella and determination of spvR gene from isolated samples of east Azarbaijan province flocks. From 45 suspected flocks overall 200 samples taken from liver and ovary. Samples was cultured in selective and differential growth medium of salmonella and OMPG was performed. Antibiogram test to determination of antibiotic sensitivity was done following isolation of bacteria. For detection of spvR gene in isolated salmonella the polymerase chain reaction with specific primer was done. PCR was used to determine the presence of spvR gene with specific primers. At first, 2 ml tubes were selected and with the TE band and added buffer then a sample of bacterial colonies was dissolved in each tube then dissolved for 1–2 minutes became vortex. The samples were heated on a hot plate for 22 minutes to boil and the lysis of bacteria release the DNA. The samples were then centrifuged and centrifuged at 10,000 rpm for 12 minutes, then the supernatant was removed (clear fluid) and placed on 2 mL tubes on the nanodrag and if the marker of 260-280 was between 1.8-2 indicates good purity and good DNA concentration. Samples were then transferred to an -80°C freezer to be ready for the PCR step. The spvR-F and spvR-R was used in this study. The results of study was analyzed statistically by SPSS version 22. The 200 samples studied, a total of 26 Salmonella isolates were isolated and the rest of the samples were negative for Salmonella bacterial culture test. The results of statistical analysis showed that 13% of samples were positive for Salmonella and the rest were negative. The 23 samples (88.46%) has the spv virulence gene. The results indicated all isolates was resistant to Erythromycin, Tetracycline, and Trimethoprim-Sulphamethoxazole, and the highest antibiotic resistance was against Doxycycline 94.3%, Danofloxacin92.6%, Florfenicol 91.7%. The highest sensitivity regard to Fosfomycin 94.7%, and Enrofloxacin 74.2%. Results of molecular tests indicated the spvR gene was existed in layer flocks of east Azarbaijan province. Researchers have shown that the spvR gene is present in 100% of isolated salmonella. In Brazil 82.7% of the samples taken after food poisoning has spvR gene. It was indicated that the Salmonella was isolated from 45.52% of poultry samples and in 14.69% of cases spvR was present. In Chaharmahal va Bakhtiari province, 160 (52.45%) out of 305 samples were infected with Salmonella and the frequency of spvB, spvC, spvR genes was 45.7, 76.6, and 69.14 percent, respectively (Daruoshi et al., 2015). Three spvB, spvC, and spvR genes were present in 60 serotypes of salmonella isolated from different sources and their frequency was 43.3, 73.3, and 46.6%, respectively. The results of researchers at a slaughterhouse in Kerman showed that out of 1001 poultry samples collected, 68 were infected with Salmonella. Overall, 88.6% of the samples also confirmed the presence of the spv gene. The results of the present study showed that 23 cases (88.5%) has spv virulence gene. This is in line with the results of previous studies in this area. Amini (2010) found that 88.6% of spv genes were present in isolated salmonellas. In Brazil, 82.7% of samples the spv genes was present (Geimba et al., 2004). However, Dariush et al., (2015) Only 14.69% of spv virulence genes were present (Daruoshi et al., 2015). The resistance rate against various antibiotics in layer flocks of east Azarbaijan was high and the virulence spvR gene exist in infected salmonellas of layer flocks.

Nowadays the life style and conditions have caused that human exposed to various stresses. Various types of physiological and psychological stresses affect the hypothalamic-pituitary-adrenal cortex, the hypothalamic-pituitary-gonadal axis, the adrenomodular sympathetic system, and the sympathetic nervous system, resulting in alterations in some of the body organs. One of the most affected systems is reproductive organs and this effect is triggered by activating a cascade of reactions. There are reports that stress with various degrees resulted to reduced testosterone secretion and may affect spermatogenesis in males. One of the strongest of stress is immobility, which results in an increase in some hormones such as adrenaline, noradrenaline and corticosterone. According to research, this type of stress in the brain increases nitric oxide production, which suppresses testosterone production. Now, medicinal plants and their derivatives account for 20 percent of prescriptions in advanced industrialized countries and 80 percent in developing countries. Anthemis nobilis L. is one of the native plants of Mediterranean region which is used in traditional Iranian medicine as a pain reliever and antispasmodic agent. Previous researches have shown that Anthemis nobilis L. can have positive effects on reproductive parameters. Considering the effects of Anthemis nobilis L. in traditional medicine as an effective medicine on some diseases, the aim of this study is evaluating the effect of hydro-alcoholic extract of Anthemis nobilis L. on parameters of reproductive and behavioural in male rats exposed to immobility stress.

30 male rates weighing 220 ± 20 g were randomly divided into five group include: sham without any stress and additive, control exposed to stress without additive, treatment groups exposed to stress and received of Anthemis nobilis L. in dosage 50, 100 and 200 mg/kg respectively. The extract was given to the mice with gavage and accompanied by immobility stress for 21 consequence days. At the end of study, anxiety was evaluated using elevated plus maze, also blood samples were collected for hormones assay and number of sperms were assessed. All procedures of elevated plus maze recorded by a digital camera and behaviors evaluated using Panlab v.3 software. Obtained data were analyzed using SPSS v.23 software under windows and to compare differences between the groups, one-way ANOVA and Tukey post-hoc test was used.

Studies showed that 50, 100 and 200 mg/kg hydro-alcoholic extract of Anthemis nobilis L. could increase the number of sperm (respectively 51.32±2.61, 54.25±3.08 and 58.11±3.16) , Testosterone (respectively 2.24±0.10, 2.40±0.09 and 2.64±0.13), LH (respectively 1.83±0.10, 1.96±0.13 and 2.1±0.12) and FSH (respectively 3.58±0.17, 3.72±0.11 and 4.12±0.12) levels compared to the stress group significantly (p

Selenium play a crucial role in the formation of selenoprotein, hormones and intrinsic factors. The stearoyl-CoA desaturase enzyme (SCD) is present in many tissues of the body and its activity leads to the formation of a single-stranded fatty acid from saturated fatty acids. Present study was carried out on fat tail weaning male lambs to evaluate the effect of oral selenium nanoparticles administration on transcription rate of stearoyl-CoA desaturase enzyme. For this purpose, 14 male lambs were selected at weaning time and divided into three groups: treatment 1 (5 heads), treatment 2 (5 heads) and control (4 heads). Selenium nanoparticles were administered for 20 consecutive days at a dose of 0.05 and 0.1 mg/kg B.W. to treatment groups 1 and 2, respectively and distilled water to the control ones. Tail fat tissue samples were collected at days 0, 20 and 30 via core biopsy needle gauge 14. Transcription rate of stearoyl-CoA desaturase (SCD) enzyme gene was performed by RT-PCR method and data were analyzed statistically using ANOVA test at the level of p

Hepatoid glands or Perianal glands are modified sebaceous glands in carnivores which they are referred to as, because of their histologic resemblance to hepatocytes (Kokila et al., 2016). In adult male dogs these glands enclose the subcutaneous perianal tissues, but in female dogs these glands creat diffuse islands in the subcutaneous perianal tissues (Wilson et al., 1979). Most hepatoid gland tumors are adenoma type. Carcinomas of these glands are malignant tumors that show more aggressive behavior than adenomas (Jardim et al., 2018). Hepatoid gland tumors are seen in uncastrated males at the age of 8 to 12 years (Wheler et al., 1996). However, lower numbers have also been observed in sterile female dogs and males (Berrocal et al., 1989). The risk of tumors in these glands in males is 5-6 times higher than in females (Atoji et al., 1989). Approximately 10% of these tumors occur ectopically in other locations, such as the tail, perineal area, prepuce, flank, thigh, back, sacra-lumbar region, midline of abdomen, cheek, head and neck (Trangadia et al., 2014). Hepatoid gland tumors are the third most prevalent among canine skin tumors, with reported frequency of benign cases about 58-96% and frequency of malignant cases about 3-17% (Vail et al., 1990; Trangida et al., 2014). Malignant hepatoid gland tumors may metastasize to the sacral and sublumbar lymph nodes. However, the risk of metastasis in these tumors is estimated 10 to 30% (Goldschmidt et al., 2002). Three types of surgery, radiation or chemotherapy are often used to treat tumors in dogs (Doustar et al, 2010). The ectopic incidence of these tumors in regard to their anatomical locations has increased the importance of this report.

In January and March of 2019, two cases of skin tumors observed in two dogs with different breeds in Isfahan Veterinary Pathology Center (private section). First Case: A male terrier dog, 18 years old and weighing 10 kg, kept at home and uncastrated. The reason for the dog's referral was due to the prominent growths on the skin of the flank and the left hand with rapidly growing. At clinical examination, the masses were firm in palpation, painless, and wounded with bleeding, about 4 to 5 cm in diameter in the flank region and about 2 to 3 cm in diameter on the left hand. Second Case: A male German shepherd dog, 10 years old and weighing 35 kg which was kept in a garden around Isfahan. The reason for the referral was also the appearance of a large, rapidly growing, wounded mass in the back of the neck and below the collar, which had a firm stiffness in the palpation. Within a month the mass had reached a diameter of about 15 cm. The animal was febrile and inappetence due to infectious nature of the tumor mass. The animal was also uncastrated. Two cases were biopsied and tissue samples in 10% neutral formalin were sent to Isfahan Veterinary Pathology Center for histopathological examination.

In histopathological examination, hepatoid gland with tissue structure similar to normal hepatoid glands was observed. The glandular tissue was divided into separate sections by fine trabeculae of connective tissue. Proliferation of hepatoid cells was observed in the gland tissue. Hepatoid cells observed with distinct border, polyhedral, round to oval with eosinophilic cytoplasm and partly granular with relatively large vesicular nucleus containing one or two prominent nuclei. Also smaller reserve cells were observed sparsely. The stroma included blood capillaries and inflammatory cells. The mitotic figures were prominent and abundant. In addition, congestion and hemorrhage (in the first case) and necrosis and infiltration of inflammatory cells especially neutrophils (in the second case) were also observed. According to histopathological findings, the lesions were diagnosed in both cases as Well Differentiated Hepatoid Gland Carcinoma. Histopathologically, hepatocellular tumors are classified into three groups: adenoma, carcinoma and epithelioma. Berrocal et al. (1989) divided the hepatoid gland tumors into four groups: well-differentiated, moderately or poorly differentiated, carcinomas without differentiation and non-characteristic perianal structures, and reported their frequency, 58.3%, 21.6%, 16.5% and 3.6%, respectively. Well-differentiated carcinomas of the hepatoid glands are similar to hepatoid gland adenomas microscopically. Therefore, careful microscopic examination and observation of hepatoid cell proliferation, reserve cells and presence of mitotic figures may indicate the malignancy of these glands (Goldschmidt et al., 2002). Development of hepatoid gland tumors is hormone-dependent (Atoji et al., 1989). Therefore several studies have been done on androgen receptors in normal and neoplastic hepatoid glands (Yamusak et al., 2016). Hepatoid gland adenomas usually respond positively to estrogen hormone therapy, so that these tumors subside following estrogen administration or castration (Wilson et al., 1979). In the female dogs, most tumors are developed following ovariohysterectomy, decrease of estrogen level and inability to suppress hepatic gland tissue growth, and in the male dogs, interstitial cell tumors lead to the elevated androgen levels in affected dogs, so the rate of hepatoid gland tumors increases (Hayes et al., 1977). However, hepatocellular carcinomas do not subside after treatment with estrogen or castration (Vail et al., 1990). Some have also suggested that dogs with hepatocellular carcinoma benefit from estrogen treatment rather than castration (Pisani et al., 2006). Although surgery is the best treatment, but in addition to surgery and extensive tumor resection, castration and anti-androgen therapy could be used in the treatment of the male dogs (Devi et al., 2012).

At this study none of the affected dogs were castrated. Therefore, as a conclusion, their old age, male sex, sexual activity and probably high androgen levels might be increased the risk of hepatocellular carcinoma in two cases. However, further studies are needed to determining the effects of these hormones on carcinogenesis of these tumors.