Veterinary Research Forum
Volume:12 Issue: 3, Summer 2021

The aim of this study was to determine the potential fertilizing of spermatozoa from the epididymal tail in different periods of time post-orchiectomy (P-OQ). Therefore, the study was approached in two stages. In the first stage, the orchiectomy was performed in 30 adult pigs. The testicles were stored at 5.00 ˚C in physiological saline solution for 5, 24, 48, 72, 96 and 120 hr. The spermatozoa were obtained by backflushing the vas deferens. The spermogram and fluorometric study were performed for each sample to evaluate the exposure of phosphatidyl-serine (PS) and acrosome reaction (AR). The second stage included the fertilization test, 16 prepubertal sows were selected, after synchronizing the oestrous cycle and the post-cervical artificial insemination was performed with the refrigerated sperm samples from each P-OQ time. The percentage of live sperm remained without significant changes until 96 hr P-OQ. An increase in the percentage of spermatozoa that showed a PS exposure was observed. The premature AR was evident after 72 hr. Considering that the artificial insemination was performed ensuring a minimum number of live sperms, no significant differences were observed in the number of embryos and corpora lutea. The results indicated that pig sperm collected from the epididymal tail P-OQ and stored for 5 and up to 72 hr at 5.00 ˚C had viable characteristics and maintained their fertilization ability. However, there was an increase in the loss of phospholipid asymmetry of the plasma membrane as time increased (72 and 96 hr), therefore, sperm viability was decreased.

The aim of this study was to investigate the presence of caprine herpes virus-1 (CpHV-1) and bovine herpes virus-1 (BoHV-1) in 269 goat sera collected from small-scale family farms located in six provinces within the Black Sea region of northern Turkey. The overall seropositivity for alpha-herpesvirus in the native goats was found as 19.33% using BoHV-1 glycoprotein B (gB)-blocking enzyme-linked immunosorbent assay (ELISA). Additionally, the seroprevalence of BoHV-1 was determined in 5.20% of the goats using virus neutralization test. To distinguish between CpHV-1 and BoHV-1, the combinations of gB/gE-blocking ELISA tests were performed. Of tested samples, 15.24% were CpHV-1 seropositive; whereas, 4.09% were BoHV-1 seropositive. The results indicated that CpHV-1 is in circulation among local goats of northern Turkey. Considering the close relationship between BoHV-1 and CpHV-1, the transmission of BoHV-1 via goats may also be one of the predisposing factors involving in the spread of virus among the surrounding cattle.

Ceruminous gland tumor is the most common tumor of the ear canal in cats. Otoscopic examination of the ear tumor is so difficult due to narrowing of the external ear canal. The present study aimed to investigate clinical, ultrasonographic and histopathological characteristics of feline ceruminous gland neoplasm in cats. Ten cats with unilateral ear canal swelling were subjected to thorough physical and clinical investigations. Ultrasound of the ear canal and parotid gland was performed using 8.00 MHz linear probe. Tissue specimens were collected after surgical excision (total ear canal ablation) for histopathological examination. Clinical examination of the ceruminous tumors revealed firm pinkish mass obliterated the ear canal with purulent or bloody aural discharge. Ultrasound examination of the ear tumor was helpful in detecting the size, shape, echogenicity and extension of the tumors to the surrounding structures as well as the nature of the feline ceruminous tumor. Histopathological examination was the main diagnostic tool for detecting the nature of the ceruminous neoplasms.

Kisspeptin has an important role in the stimulation of hypothalamic-pituitary-gonadal axis in term of pubertal development, release of reproductive and metabolic hormones and ultimately affecting the fertility. The aim of the present study was to evaluate the serum kisspeptin level and its correlation with semen quality and selected hormones in buffalo bulls during the summer and spring seasons. Semen and blood samples from eight Nili-Ravi buffalo bulls (age: 9.21 ± 1.02 years) were collected. Semen was analysed using computer-assisted semen analysis. Serum concentrations of kisspeptin, gonadotropin releasing hormone (GnRH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone, cortisol, triiodothyronine (T3), thyroxin (T4) and insulin like growth factor (IGF-1) were estimated using enzyme-linked immunosorbent assay kits. Kisspeptin was neither affected by seasons and nor correlated with semen parameters and hormones. Higher levels of GnRH, LH, cortisol, IGF-1, total motility (TM), average path velocity (VAP), straight-line velocity (VSL), curvilinear velocity (VCL) and linearity (LIN) were recorded in summer compared to spring. Correlations of GnRH versusIGF-1 and LH, LH versus IGF-1 and cortisol, FSH versus T4 and testosterone, testosterone versus T3 and T4 and T3 versus T4 were observed. The GnRH and IGF-1 were positively associated with TM, VAP, VSL, VCL and LIN. The LH was correlated with VSL, straightness and LIN. In conclusion, GnRH, LH, and IGF-1 correlations with semen parameters can be used to indicate semen quality. The buffalo bulls are well-adapted and can give quality semen in the summer season.

Fasciolosis is a zoonotic parasitic disease caused by the trematode Fasciola hepatica. The proteases are essential for the survival of parasites. The present study was aimed to determine serine proteases activities in miracidia of F. hepatica and evaluate the effects of pH and different inhibitors on the serine proteases activities. Adult F. hepatica helminths were removed from naturally infected livers of the slaughtered cattle and crushed. The eggs were incubated at 28.00 ˚C for 16 days. The released miracidia were homogenized and total proteolytic activity of the extract of miracidia at different pH values were evaluated. Serine proteases activities were determined using specific substrates. The inhibitory effects of chemical and herbal inhibitors on the enzymes were also assessed. The extract of miracidia hydrolyzed azocasein with optimum activity at pH 8.00. The optimum pH effect on serine proteases activities was found at alkaline pH. Phenylmethylsulfonyl fluoride and Bowman-Birk inhibitors inhibited and decreased the proteases activities in the miracidia extract. It was concluded that there were proteases activities in miracidia of F. hepatica which were inhibited by chemical and herbal inhibitors.

The objective of the study was to determine the effects of conjugated linoleic acid (CLA) supplement on milk yield and composition, blood metabolites and reproductive parameters in lactating Holstein dairy cows. Twenty Holstein dairy cows were randomly assigned to one of two dietary treatments: 1) supplementing 110 g per day of fat (control), 2) supplementing 120 g per day of rumen-protected CLA. The diets were formulated to be nutritionally isocaloric and isonitrogenous. The experimental period started 21 days pre-calving and continued until 60 days in milk (DIM). Treatments had no effect on dry matter intake (DMI), body weight (BW) and body condition score (BCS). The CLA treatment increased milk yield (3.04 kg per day and milk lactose concentration, but decreased milk fat concentration and, short and medium chain fatty acids concentrations. No treatment differences were observed in milk protein concentration, milk energy output and net energy balance. Serum concentrations of glucose, cholesterol, triglyceride (TG), insulin, insulin-like growth factor-1(IGF-1), estradiol and progesterone were higher in CLA treated cows when compared to cows fed on the control diet. Serum beta-hydroxybutyric acid (BHBA) concentration was reduced in cows fed on the CLA treatment. Days to first insemination and days open were not different between the two treatment groups. Cows fed on the CLA supplement had increased conception rate from the first service. The results indicated that cows fed on diets supplemented with CLA produced milk with decreased milk fat concentration whereas some related cow blood serum metabolic parameters associated with reproductive response were increased and resulted in an increased conception rate from the first service.

The purpose of this study was to investigate the effect of two aflatoxins (AFs) sources for experimental induction of aflatoxicosis in ducklings. Dietary supplementation of aflatoxin-contaminated grounded rice grains previously was cultured by Aspergillus parasiticus and dietary supplementation of aflatoxin methanolic extract of contaminated rice grains. A total number of 18 four-day-old ducklings were treated with varying sources of AFs. Treatments included: A: Control (basal diet without AFs), B: Contaminated feed with 0.20 mg kg-1 AFs (ground rice grains), C: Contaminated feed with 0.20 mg kg-1 AFs (methanol extract of contaminated rice grains). Results showed a significant decrease in serum levels of total protein, albumin, glucose, triglyceride, cholesterol, HDL, LDL and creatinine. The serum's hepatic enzymes  levels were not affected in AFs-treated groups but lactate dehydrogenase (LDH) concentration increased by the two AFs sources. The relative weight of the liver and spleen was significantly increased in AFs-fed groups. Histopathological liver examination showed the vacuolar degeneration with small and large lipid droplets in hepatocytes in the AFs- contaminated groups. Dietary AFs resulted in a significant decrease of villus height, villus width and villus surface area of the small intestine compared to the control group. In conclusion, the results showed that the induction of experimental aflatoxicosis via two investigated AFs sources had slight differences concerning the studied parameters. It seems the group consumed ground rice grains indicated slightly fewer aflatoxicosis symptoms than the methanolic extract.

Gastrointestinal motility disorders can occur as either increased or decreased movements. Studies have shown that herbal ingredients such as essential oils can modify the increase and decrease of gastrointestinal movements of ruminants. Cinnamaldehyde at room temperature is an oily yellow liquid which is obtained from the steam distillation of the oil of cinnamon bark. It bears carminative activity and gastrointestinal, antimicrobial, and vasodilatory effects. This study examined the effects of cinnamaldehyde on the contraction of circular smooth muscles of abomasal fundus and the antrum, duodenum, and ileum of healthy cows using an in vitro approach. The results indicated that cinnamaldehyde had relaxant effects on the basal tonus and contractions caused by barium chloride (BaCl2) and carbachol (CCh) in these tissues dependent upon concentration and the origin of the smooth muscle. These effects were more prominent in the ileal smooth muscle preparations than in other tissues. This substance in the smooth muscle preparations of the abomasal fundus not only had no significant effect on the basal tonus, but also significantly increased the contractions caused by barium chloride at low concentrations. Study of the mechanism of action showed that, similar to verapamil, cinnamaldehyde applied its relaxation effect by blocking the calcium channels. The results showed that cinnamaldehyde possessed a spasmolytic effect mediated through blockage of the calcium channels, which may provide a pharmacological base to its medicinal use for diarrhea and spasms.

Picobirnavirus (PBV) is an enteropathogen virus causing diarrhea as an opportunistic virus in its vertebrate host. There is no information about human or animal PBVs in Iran. The aim of the present study was the investigation of the epidemiology of bovine PBV in the broad geographical area of Iran. Four hundred and eighty-five stool samples of up to 1 month old diarrheic calves were collected from 14 provinces and were tested with polyacrylamide gel electrophoresis (PAGE), and reverse transcription polymerase chain reaction (RT-PCR). Five samples were positive in PAGE assay (1.00%) and all of them were amplified using GI specific primers in RT-PCR. Phylogenetic analysis of one of the amplicons (strain Nazaktabar-14) revealed a low relationship to bovine PBV sequences and more identity to PBV isolates from other hosts. The structural alignment of the deduced amino acids of the partially sequenced RdRp gene of the Nazaktabar-14 strain showed high conservation. Sequences obtained from other amplicons showed a high mutation rate and further analysis of one of them showed that, despite the potential of forming deleterious mutations, most of the point mutations occurred in the RdRp gene of PBVs may be a silent mutation. There is little information about the molecular epidemiology of bovine PBVs. This study was the first report on the occurrence of PBVs in Iran and the first study on the molecular epidemiology of bovine PBV in the Middle East, revealing its low frequency as a diarrhea causative agent.

To investigate the hormonal and biochemical profiles of horses with osteochondrosis dissecans (OCD), serum insulin, cortisol, triiodothyronine, thyroxine, fasting blood glucose (FBG), cholesterol, triglyceride (TG), high- and low-density lipoproteins, albumin and uric acid were measured in horses definitely diagnosed with OCD (n=19) as well as clinically normal horses (n=18). Proxies representing insulin sensitivity [reciprocal of square root of insulin concentration (RISQI)] and beta cell responsiveness [modified insulin to glucose ratio (MIRG)] were calculated. Body fat percent (BF%) was estimated according to fat depth over the rump using ultrasonography. Body condition score (BCS), weight, and waist circumference were also determined. Glucose was significantly higher and MIRG, BCS, BF% and TG were significantly lower in OCD- horses compared to control group. Based on BCS scores, horses in control group were overweight. The results of the present study, higher FBG and lower MIRG, might implicate the existence of a footmark of insulin/glucose derangement. The body mass index and muscle mass were not measured in this study; nonetheless, a lower BF% might implicate a higher body muscle mass in OCD affected horses, which were comparably underweight compared to control group. While insulin resistance does also occur in human individuals and horses with lower BF%, horses with higher muscle mass may show greater potential for exercise, which in turn, exerts greater physical pressure on cartilages. An underlying hormonal predisposition could make these horses more prone to OCD, originally triggered by mechanical pressures.

Ethephon (C2H6ClO3P; ETP), an organophosphorus pesticide regulating plant growth, is widely used for early ripening of fruits and vegetables in agriculture. The aim of this study was to investigate the effects of ETP on histomorphometrical and biochemical parameters in mouse testicular tissue. In this study, 90 adult male mice were randomly divided into six equal groups (n = 15). The ETP was administered orally at different doses (120, 240 and 480 mg kg-1) daily for 35 days. Untreated control, sham (received only normal saline) and neostigmine bromide-treated (positive control; 0.10 mg kg-1 orally; once per week) groups were also considered. Following 35 days, animals were euthanized and testicle and serum samples were taken. Accordingly, blood and serum acetylcholinesterase (AChE), catalase, total antioxidant capacity (TAC) and malondialdehyde (MDA) levels as well as histomorphometrical changes of testicles were investigated. The ETP-administered animals represented a significant reduction in AChE, TAC and catalase levels and remarkable increment in MDA content. A marked reduction was also seen in the germinal epithelium height, connective tissue thickness, seminiferous tubules diameter and Leydig cell number as well as spermiogenesis and Sertoli cell indices in ETP-treated mice compared to control ones. Similar findings were found in neostigmine bromide-treated animals. In conclusion, the ETP significantly affects the serum and blood anti-oxidant statuses and results in severe histological damages both at germ and somatic cell levels, suggesting its hematotoxic and reprotoxic characteristic.

The aim of this study was to acquire an effective method for preparation of rat decellularized kidney scaffolds capable of supporting proliferation and differentiation of human adipose tissue derived mesenchymal stem cells (AD-MSCs) into kidney cells. We compared two detergents, the sodium dodecyl sulfate (SDS) and triton X-100 for decellularization. The efficiency of these methods was assessed by Hematoxylin and Eosin (H&E), 4', 6 diamidino-2-phenylindole and immunohistochemistry (IHC) staining. In the next step, AD-MSCs were seeded into the SDS-treated scaffolds and assessed after three weeks of culture. Proliferation and differentiation of AD-MSCs into kidney-specific cell types were then analyzed by H&E and IHC staining. The histological examinations revealed that SDS was more efficient in removing kidney cells at all-time points compared to triton X-100. Also, in the SDS-treated sections the native extracellular matrix was more preserved than the triton-treated samples. Laminin was completely preserved during decellularization procedure using SDS. Cell attachment in the renal scaffold was observed after recellularization. Furthermore, differentiation of AD-MSCs into epithelial and endothelial cells was confirmed by expression of Na-K ATPase and vascular endothelial growth factor receptor 2 (VEGFR-2) in seeded rat renal scaffolds, respectively. Our findings illustrated that SDS was more effective for decellularization of rat kidney compared to triton X-100. We presented an optimized method for decellularization and recellularization of rat kidneys to create functional renal natural scaffolds. These natural scaffolds supported the growth of AD-MSCs and could also induce differentiation of these cells into epithelial and endothelial cells.

Echocardiography is a non-invasive method of cardiac evaluation in most species. Echocardiographic reference parameters are poorly documented in sheep and not documented in Ghezel sheep. The purpose of this study was to determine reference ranges of normal echocardiographic parameters using two-dimensional and M-mode techniques in Ghezel sheep. In 15 healthy female Ghezel sheep aged between 15 and 18 months, echocardiography was performed in standing position from left and right parasternal approach focused at 3rd - 5th intercostal spaces using a 2.50 - 5.00 MHz phased array transducer. The following parameters were measured in two-dimensional echocardiography: left atrial diameter (LAD), mitral valve annulus (MVA), aortic sinus (AoS), aortic valve (AoV), pulmonary sinus (PuS), and pulmonary valve (PuV); and in M-mode echocardiography: interventricular septum (IVS), left ventricular internal diameter (LVID), left ventricular free wall (LVFW), right ventricle free wall (RVFW), right ventricle internal diameter (RVID). Calculated variables included the ratios LAD/Ao and Pu/AoS, and the fractional shortening (FS), end diastolic volume (EDV), end systolic volume (ESV), ejection fraction (EF), stroke volume (SV) and cardiac output (CO) of the left ventricle. In conclusion, echocardiographic parameters could be reliably assessed in Ghezel sheep and our study provided some normal echocardiographic reference ranges that might be useful in proper identification, visualization, and measurements of cardiac structures. Such findings could be useful to assess and to diagnose the specific heart diseases in sheep practice and also for experimental studies in sheep as an animal model used for research purposes in cardiovascular studies of human.

Cardiac problems are highly fetal diseases among exotic animals, not only in the rabbit which is prone to such diseases but also in the guinea pig (Cavia porcellus). In rodents, imaging studies such as thoracic radiography are more practical and easier to perform than echocardiography. Cardiac size is primarily evaluated using vertebral heart size (VHS) as reported in ferrets and rabbits. We therefore attempted to determine standard cardiac dimensions in the guinea pig by thoracic radiography using VHS. The purpose of this research was getting an indicator of the normal range of male and female, mature and apparently healthy guinea pigs heart. Standard radiographs of the thorax in lateral and ventro-dorsal (VD) views were taken and interpreted. In our study to determine VHS in VD view conventional method was used. In addition to conventional VHS method, two other measurement methods were performed in lateral view. Statistical analyses were performed with a SPSS Software and Mann-Whitney U test to compare results. Mean and standard deviation were also calculated. According to the results, the total average of VHS in lateral view by the first method was 7.80 ± 0.12, by the second method was 7.80 ± 0.16, by the third method was 7.60 ± 0.15 and the total of average of VHS in VD view was 9.20 ± 0.23. According to present research findings, gender had no meaningful effect on the measuring of the heart size. More researches on the same age and the same species guinea pigs are needed for more accurate evaluation.

Mugger crocodile is the only crocodile existing in Iran. The present study was aimed to investigate the bacterial flora in oral and cloacal cavities of wild Mugger crocodiles in Negour protected area, Iran. The isolation and molecular characterization of oral and cloacal bacterial flora were performed in 22 Mugger crocodiles captured in Negour protected area, Iran. Ten bacterial species from all oral samples and six bacterial species from all cloacal samples were recovered. The most commonly isolated bacteria in oral samples were Burkholderia contaminans and Lactococcus garvieae, respectively; whereas, in cloacal samples, it was Lactococcus lactis. It is likely that the isolated bacteria would pose a threat to both crocodiles and humans health. It can threaten crocodiles during stressful conditions; while, humans would be susceptible if they are bitten by crocodiles, consume their meat or spend time near their natural environment. This study provides useful information about bacterial diversity which could help to select the most appropriate anti-bacterial when dealing with infections caused by crocodiles.

The effects of garlic oil (GO) on serum fatty acids, lipid profiles and energy-related blood metabolites in feed-restricted fat-tailed ewes were investigated. For this purpose, twenty-eight dry, non-pregnant Makuei ewes (about three years of age; 45.00 ± 2.20 kg of body weight) were assigned randomly to four experimental groups including group 1 receiving basal diet as total mixed ration (TMR) without GO supplementation (control group), group 2 receiving 100% basal diet supplemented with GO (10.00 mg kg-1 BW), group 3 receiving 70.00% basal diet without GO supplementation and group 4 receiving 70.00% basal diet supplemented with GO (10.00 mg kg-1 BW). The main experimental period started eight weeks after performing adaptation and dietary allocations. Feed restriction reduced serum glucose levels along with higher serum levels of non-esterified fatty acids, triacylglycerols, β-hydroxybutyrate, low-density lipoprotein cholesterol, total cholesterol and very low-density of lipoprotein. Following feed restriction, the serum palmitic and oleic acids concentrations were increased. Garlic oil supplementation had a desirable effect on feed-restricted animals through lowering serum BHB and palmitic and oleic acids concentration and increasing the high-density lipoprotein cholesterol levels in the serum. These findings demonstrated that GO had the potential to reduce body-fat mobilization, thereby lowering the risk factors for disorders associated with negative energy balance in underfed ewes in the periparturient period.

Paramphistomosis is an infectious disease caused by the liver flukes and it is associated with heavy loss of ruminant's population. Explanatum explanatum is a digenetic trematode commonly affecting domesticated ruminants. The available methods for pathogen detection are laborious and expensive and offer limited specificity; thus, considered not suitable for post mortem pathogen detection, surveillance and prevalence studies. New detection techniques offering simplicity, specificity and rapidity are absolutely needed. We have designed a loop-mediated isothermal amplification (LAMP) based polymerase chain reaction method, targeting a sequence of the explanatum species, using a primer pair from the internal transcribed spacer-2 region. The DNA from adult flukes belonging to explanatum species was isolated from infected livers and used to optimize the LAMP assay. The specificity and sensitivity of the LAMP assay were evaluated and found highly efficient in species-specific DNA detection with the sensitivity to detect 50.00 pg DNA in a 25.00 µL reaction mix. The procedure has the potential to be adapted for stool samples for field detection and disease surveillance/prevalence in rural and unprivileged areas.

The purpose of the present study was to evaluate effects of medetomidine on left ventricular outflow tract (LVOT) velocity in domestic short-haired cats. Eighteen healthy adult male domestic short-haired cats were used for this study. All animals were client-owned. Echocardiography machine with 7.50 MHz transducer was used. Specific veterinary two-dimensional and pulse-waved echocardiogram images in apical five chamber right parasternal view were obtained and blood velocity in LVOT was calculated. After baseline echocardiographic recordings, 0.04 mg kg-1 of medetomidine was intramuscularly administered to each animal and LVOT velocity was calculated after 15 (T15), 50 (T30) and 80 (T80) min following drug administration. The LVOT velocity values (mean ± SEM) of cats in baseline were 1.06 ± 0.04 m sec-1. There were significant differences between baseline and T15 and T30 regarding mean LVOT values. Age and weight had no significant effect on LVOT velocity values. The LVOT velocity values of T15, T50 and T80 were 0.77 ± 0.04, 0.80 ± 0.02 and 0.96±0.03 m sec-1, respectively. Our findings revealed significant decrease in mean LVOT velocity up to 50 min following medetomidine administration. The present study determined normal LVOT velocity range for a small population of cats before and after intra-muscular medetomidine administration.

One of the most critical emergencies in dogs is esophageal foreign bodies. The most important foreign body reported in the esophagus is bone; however, there are also reports of fish hooks lodged in the esophagus. This case report describes the history, clinical signs, radiographic findings and surgical management of an esophageal foreign body due to a single-barbed fish hook in a 4-month-old Siberian Husky female dog with regurgitation, pain, anorexia and lethargy. In this case, plain radiography was taken to determine the type and location of the foreign body and it was confirmed that a single-barbed fish hook was lodged in the heart base of thorax. Depending on the type of foreign body, penetrable or non-penetrable as well as the location, a variety of methods including endoscopy, fluoroscopy, and surgery are used for its removal. For treatment, for the first time, a covault hook was utilized to remove the foreign body via gastrotomy. Case follow-up during two weeks showed no postoperative complications and the patient was healthy.

Ocular tumors are rarely seen in farm animals. Iridociliary epithelial tumors are the second most common tumor of the eye in dogs and cats, but there is just one report of this tumor in sheep. This case report described the history, clinical signs, histopathological and immunohistochemical findings, and surgical management of an iridociliary adenoma tumor in a 3-year-old Iranian Shaal sheep with signs of anorexia, depression, loss of vision, and a mass covering the left eye. The mass had protruded from the left eye globe but had not penetrated the periocular tissues. Enucleation was performed to excise the tumor. Periodic acid–Schiff (PAS) and Alcian blue staining methods were done for the mass, but the result for both of them was negative. Slides were also immunostained for antibodies against cytokeratin AE1/AE3, vimentin, and S-100 protein. The results were positive for vimentin but negative for cytokeratin and S-100. According to the results of histopathology and immunohistochemistry, the type of tumor was diagnosed as iridociliary adenoma. To our knowledge, it was the second report of the iridociliary tumor in the sheep and the first report of this tumor in the Iranian Shaal breed.