Veterinary Research Forum
Volume:12 Issue: 2, Spring 2021

Harris hawks (Parabuteo unicinctus) are the raptors with more anthropogenic use, which makes it necessary to improve their reproduction either natural or assisted for sustainable use even though they are not endangered. The aim of this study was to determine fresh and thawed P. unicinctus sperm capacitation and acrosome reaction parameters as indicators of sperm viability in ejaculates of birds with injuries. A total of 30 ejaculates were analyzed as fresh and thawed samples. Basic sperm evaluations as well as in vitro acrosomal reaction capability through the presence and distribution of Ca2+ and N-acetylglucosamine and sialic acid using chlortetracycline and agglutinin-fluorescein isothiocyanate Triticum vulgaris were performed,respectively. Tagged samples were observed using epifluorescence microscopy. The results indicated 30.00% decrease in sperm viability following freeze-thaw, while rate of capacitated sperms (10.00%) and the percentage of sperm with acrosomal reaction (40.00%) were increased. The results of the experiment demonstrated that the post-thaw viability of the obtained sperm may be an alternative for use in artificial insemination.

This study aimed to assess the superovulation response and pregnancy rates of fresh and vitrified-thawed embryos after transfer in Angora goats with comparing transfer at the beginning (BS) and end (ES) of the breeding season. Nine Angora goats were used as donors in both periods. Donor goats were synchronized and superovulated with the FSH and mated with five fertile bucks. At 156 hr following mating, embryos were collected surgically. Recipient Angora goats were divided into two groups at the beginning (fresh, n=15; vitrified-thawed, n=15) and end (fresh, n = 8; vitrified-thawed, n = 8) of the breeding season. Fresh or vitrified-thawed grade I embryos (early blastocyst/blastocyst) were transferred surgically to synchronized recipients. On the 30th, 60th and 90th day of transfer, goats were examined by ultrasonography. The number of corpora lutea (CL), total oocyte/embryo and transferable embryos obtained in BS was found to be statistically higher than ES. On the 30th day of transfer, pregnancy rates were 73.30% and 75.00% in the fresh transfer groups in both BS and ES periods; while, rates of 20.00% and 37.50% were found in the vitrified-thawed group, respectively. The embryo survival rates of fresh transfers were 55.55% and 31.25% at BS and ES, respectively. The number of CL, total oocyte/embryo and transferable embryos in the BS was higher than ES following superovulation. Also, fresh embryo transfer can be successfully carried out during the breeding season in Angora goats. Moreover, although pregnancies were obtained following the transfer of vitrified-thawed embryos, they did not sustain on the 60th and 90th days. So, further studies are needed for the vitrified-thawed embryos.

This study was designed to investigate the effects of peripheral [intraperitoneal (IP)] and central [intracerebroventricular (ICV)] administration of cinnamaldehyde on concentrations of blood glucose and serum insulin in the acute hyperglycemia induced by ketamine/xylazine. Yohimbine (a α2-adrenoceptor antagonist) was used alone and in combination with cinnamaldehyde to explore the α2-adrenergic receptor contribution. A total of 48 rats were divided into eight groups with six rats in each for IP administration of normal saline, vehicle, cinnamaldehyde (25.00, 50.00 and 100 mg kg-1), yohimbine (0.50 and 2.00 mg kg-1) and cinnamaldehyde plus yohimbine. These rats were used again for ICV administration 15 days after the completion of IP experiment. During this 15 days period, the lateral ventricle of the brain was surgically cannulated for ICV administration of normal saline, vehicle, cinna-maldehyde (25.00, 50.00 and 100 µg per rat), yohimbine (5.00 and 20.00 µg per rat) and cinnamaldehyde plus yohimbine. Blood glucose levels were measured from tail blood using a glucometer and serum insulin concentrations were determined via enzyme-linked immune-sorbent assay kit. The increased levels of blood glucose and the decreased concentrations of serum insulin were significantly decreased and increased, respectively, by separate and combined IP and ICV administrations of cinnamaldehyde and yohimbine. The systemic effects of these chemical compounds were significantly greater than the central ones. Based on the results, it can be argued that cinnamaldehyde has a potential to induce anti-hyperglycemic and antihypoinsulinemic effects. Peripheral and central α2-adrenegic receptors might be involved in these effects of cinnamaldehyde.

Newcastle disease (ND) is known as the most common diseases of economic importance worldwide. Vaccination against virulent strains of Newcastle disease virus (NDV) has failed during some outbreaks. Here, we aimed to assess the epitopes of NDV fusion protein as targets for a peptide-based vaccine. To explore the most antigenic epitopes on the F protein, we retrieved virulent strains of genotype VII from National Center for Biotechnology Information (NCBI). Linear and conformational B-cell epitopes were identified. Moreover, T-cell epitopes with high and moderate binding affinities to human major histocompatibility complex (MHC) class I and class II alleles were predicted using bioinformatics tools. Subsequently, the overlapped epitopes of B-cell and MHC class I and MHC class II were determined. To validate our predictions, the best epitopes were docked, to chicken MHC class I (B-F) alleles using the HADDOCK flexible docking server. Seven ‘high ranked epitopes’ were identified. Among them, ‘LYCTRIVTF’ and ‘MRATYLETL’ showed the highest scores. The other five epitopes including LSGEFDATY, LTTPPYMALK, LYLTELTTV, DCIKITQQV and SIAATNEAV obtained very encouraging results as well. SIAATNEAV had been recognized as a neutralizing epitope of F protein using monoclonal antibodies before. Taken together, our results demonstrated that the identified epitopes needed to be tested by in vitro and in vivo experiments.

Histomorphometry and use of the fast free of acrylamide clearing tissue (FACT) protocol were studied on the sciatic nerve in chicken (Gallus domesticus). In the first part of the study, the sciatic nerves of 20 chickens of four age groups (7, 14, 26 and 40 days) were studied (n=5 birds per age class). Their sciatic nerve samples were stained with Hematoxylin and Eosin and Masson's trichrome and were histomorphometrically evaluated. In the second part of the study, FACT protocol was applied on the sciatic nerve of a 26 days old chicken. After clearing of 1.00 mm-thick sciatic nerve sections, they were immunolabelled using Hoechst for nuclei staining and recorded by a Z-stack motorized fluorescent microscope. In the conventional histo-morphometry, the epineurium, perineurium and endoneurium were thicker and the nerve bundle diameter was bigger in the left sciatic nerve of chicken of all age groups compared to the right sciatic nerve. On the contrary, the axon diameter and the myelinated nerve fiber diameter were bigger, the myelin sheath was thicker, the nodes of Ranvier intervals were higher and the density of myelinated nerve fibers was also higher in the right sciatic nerve compared to the left one. In conclusion, histomorphometric parameters in the left and right sciatic nerve during chicken growth were significantly different. Furthermore, the FACT protocol could be used for the 3D imaging of the chicken sciatic nerve and its immunostained evaluation.

The present study was conducted to evaluate the effects of different doses of haloperidol (HP) on induction of oxidative stress in blood and liver cell degeneration in comparison with influences of HP pre-treatment on inflammatory process induced by intraperitoneal (IP) administration of lipopolysaccharide (LPS). One hundred twenty male albino Wistar rats were randomly divided into eight groups (15 in each), including: Control group, LPS group, three groups as HP administration in three divided doses (0.50, 1.00 and 2.00 mg kg-1), and three treatment groups that HP was administered in three doses (0.50, 1.00 and 2.00 mg kg-1) prior to LPS administration.Concentrations of malondialdehyde, activities of antioxidant enzymes including glutathione peroxidase, superoxide dismutase and also the levels of tumor necrosis factor-alpha and interleukin 1-beta were measured in blood and serum. In addition to liver histopathological changes evaluation, hepatic silent information regulator of transcription 1 (SIRT1) and phosphorylated-nuclear factor-κB (p-NF-κB) levels were quantitated. Our findings indicated that sole administration of HP (particularly higher doses) can induce oxidative stress in blood and cell degeneration in liver, while it can attenuate inflammatory process induced by LPS administration presumably via SIRT1 up-regulation and preventing the induction of p-NF-κB. The oxidative and degenerative effects of HP and its impact on inflammatory status were completely dose- dependent according to our results. The possible anti-inflammatory effects of HP may affect reparative mechanisms and hepatic cell degeneration. However, the influences of HP on immune system need further investigations and its higher doses should be administered cautiously especially in patients with immune system dysfunctions.

The saffron petalsare a by-product part of the saffron flower with a cardiovascular effect. This study evaluated the effect of the saffron petalon hypertension induced by angiotensin II (AII) and NG-nitro-L-arginine methyl ester (L-NAME, a NOS inhibitor). Rats were divided into 11 groups: 1) Control, 2) AII (50.00 ng kg-1), 3) Losartan+ AII, 4) L-NAME (10.00 mg kg-1), 5) sodium nitroprusside (SNP) + L-NAME, 6, 7) Saffron petals extract; 8, 9) saffron petals (100 and 200 mg kg-1) + AII and 10,11) saffron petals (100 and 200 mg kg-1) + L-NAME. Hypertension induced by intravenous injection of AII and L-NAME in separate groups. In treated groups, 30 min before injection of AII or L-NAME rats received two doses of extract via intraperitoneal administration. The femoral artery was cannulated and cardiovascular parameters recorded by a transducer connected to power lab apparatus. Maximal changes (∆) of mean arterial pressure (MAP), systolic blood pressure (SBP) and heart rate (HR) from baseline were calculated and compared to with those in hypertensive and control groups. Results showed that both AII and L-NAME significantly increased SBP and MAP than control, however, HR in AII was decreased and in the L-NAME group increased. Pre-treatment with saffron petals could significantly attenuate the cardiovascular responses induced by both AII and L-NAME. However, the effect of the extract in AII hypertensive rats was more effective than L-NAME groups. The findings showed that the hydroalcoholic extract of the saffron petals had an antihypertensive effect that mainly was mediated by inhibition of AII activity.

Obtaining information about the eye arterial supply in the buffalo can be a criterion for ocular diseases diagnosis and give a morphological basis for even more researches on the related clinical surgery of this animal. The aim of the present study was to describe the course and branches of the arteries supplying the eye in the adult river buffalo (Bubalus bubalis). In this study, the heads of eight buffaloes were prepared from the public slaughterhouse and dissected to study the eye arterial supply. This research results showed that the eye blood supply in the buffalo originates from the external ophthalmic, internal ophthalmic, superficial temporal and malar arteries. The external ophthalmic artery was given rise to branches to take part in the ophthalmic rete mirabile formation and also given off lacrimal, external ethmoidal, supraorbital and ciliary arteries as well as muscular branches to supply the eye ball and extra-ocular muscles. The internal ophthalmic artery was one of the sources of lateral posterior long ciliary artery. The superficial temporal artery was detached off some branches to supply the lateral angle of inferior and superior eyelids as well as lacrimal branch to supply lacrimal gland. The malar artery was originated from the infra-orbital artery and its branches were supplied the medial angle of inferior, superior and third eyelids. There were no obvious differences between buffalo and yak regarding arteries supplying the eyes. However, there were minor differences between buffalo and oxen, sheep and goat and there were obvious differences between buffalo and camel and giraffe.

There has been a prevailing trend in the application of herbal medicine as cancer therapeutics. Calotropis procera is an ayurvedic plant applied to ameliorate various illnesses. There is no report on the anti-tumor effects of the root of the plant on canine tumors, although it has been used for the treatment of various diseases in human medicine. The objective of the present study was to investigate the antitumor potential of ethanolic root extract of C. procera against canine mammary tumor cell line (CF41-Mg). MTT, western blot, and flow cytometry assays were carried out to evaluate the possible cytotoxicity and apoptosis induction of the extract. MTT results showed that the extract had a potent cytotoxic activity in a dose-dependent manner with an IC50 of 9.00 μg mL-1. Based on the results of flow cytometry and western blotting, IC50 concentration of the extract induced significant apoptosis in the studied cell line, possibly through down-regulation of Bcl-2 expression. The results of the present study clearly indicated that the root extract of C. procera had promising anti-cancer activity and could be considered as a candidate for the treatment of mammary tumors.

Autoimmune diabetes is one of the most common metabolic diseases with increasing prevalence in the past decades in which pancreatic Langerhans β cells are destroyed and lead to lack of insulin due to increased blood sugar. One of the consequences of diabetes is glomerular disease of the kidney, also called diabetes nephropathy. Different studies have been carried out on the effects of triterpenoids and their medicinal effects on diabetes mellitus. betulinic acid, a pentacyclic triterpenoid of Terpenes, is found in bushes and trees. Its medical effects are also approved by many studies. In this survey, we studied the effect of betulinic acid on diabetic inbred C57BL/6 male mice. They were randomly divided to three groups. Group A: Consisted of healthy mice which received citrate buffer. Group B: Diabetic mice without any treatment and group C: Treated diabetic mice with betulinic acid. The level of blood insulin level, fasting blood glucose, C-peptide, TNF-α, IFN-γ, and IL-1 cytokines were measured and pathologic studies of the kidney were performed. The results showed that betulinic acid could increase insulin and C-peptide, and decrease fasting blood sugar, kidney lesions and TNF-α, IFN-γ, IL-1 in the treated groups. The differences were significant except for IL-1. Betulinic acid through reduction of inflammatory cytokines could have positive effects on inflammatory and autoimmune disease including autoimmune diabetes.

This study was conducted to compare the efficacy of different feed additives as mycotoxin binders in vitro. Four prevalent aflatoxin-sequestering agents (SAs) including two bentonite clays (common and acid activated bentonite), a yeast cell wall product and an activated charcoal product were evaluated in vitro to verify their capacity for binding aflatoxin B1 (AFB1). The SAs were individually mixed at two different ratios with AFB1 (1:70,000, 1:120,000) and their binding capacity indices were determined. Experimental bentonites showed high adsorption abilities, binding more than 70.00% of the available AFB1. At the 1:70,000 and 1:120,000 aflatoxin binder (AF:B) ratios, acid activated bentonite were sequestered over 87.00 and 99.00% of the AFB1, respectively. Yeast cell wall showed moderate adsorption ability at the 1:120,000 AF:B ratio, adsorbing 47.00 of AFB1. The adsorption ability of activated carbon at two AF:B ratio and yeast cell wall at 1:70,000 AF:B ratio were significantly lower than other binders. The ratio of chemisorption and binding equivalency factor were higher for acid activated bentonite compared to other sequestering agents. Based on the result of this study, it seems that acid activated bentonite could be considered efficient at sequestering the available AFB1, resulting as promising agents for use in animals diet.

One of the side effects of cyclophosphamide (CP) is low fertility. In this study, we investigated the protective role of crocin (Cr) against CP chemotherapy-induced changes in ovarian tissue. In the current study, we treated 15 female mice aged 6-8 weeks old for 21 days. The mice were distributed into three groups including control received normal saline (0.10 mL; IP), CP or sham-control group (CP once a week, 15.00 mg kg-1; IP) and experimental (CP + Cr) group received CP along with Cr (200 mg kg-1 daily; IP). After completing the procedure, levels of total anti-oxidant capacity (TAC), superoxide dismutase (SOD) and sex hormones in serum as well as malondialdehyde (MDA) in the left ovarian tissue were measured. The right ovaries were used for histological and morphological tests. The obtained data were statistically analyzed by SPSS software using ANOVA and Tukey follow-up studies. Results showed that in the CP group a significant decrease was observed in ovarian follicles, the number of corpus luteum, levels of TAC, SOD and sex hormones; while, there was a significant increase in the number of atretic follicles and mast cells and level of MDA compared to control group. Administration of Cr along with CP caused a significant ameliorative effect on the studied parameters. In conclusion, the Cr could significantly decrease the side effects caused by CP chemotherapy in mice ovarian tissue.

Bovine leukemiavirus (BLV) is one of the most important carcinogenic viruses genetically related to the human T-cell lymphotropic viruses (HTLV-1 and HTLV-2). The virus infects type B lymphocytes and creates lymph glands tumors. Recently, the association between the presence of this virus and breast cancer has been addressed in humans. Here, we studied the prevalence of BLV in the samples of raw milk of native Iranian and Iranian-foreign cows in traditional, semi-industrial and industrial dairy farms in rural and urban areas of Zanjan province. Raw milk samples of cows were collected manually in sterile tubes. The samples were tested by nested-PCR method. Forty samples (9.93%) out of 403 samples showed BLV contamination. In this study, nested-PCR was successfully applied to determine the level of contamination in raw milk samples from cows infected with BLV. Furthermore, a relatively high rate of BLV infection was found in dairy cows in Zanjan province, northwestern of Iran.

Parasitic nematodes of ovine abomasum are of economic and hygienic importance throughout the world and Iran. This study was aimed to evaluate molecular identity and species diversity of parasitic nematodes in the abomasum of slaughtered sheep in Ilam, Iran. In this study, a total number of 240 of abomasa were randomly collected from the slaughtered sheep at industrial slaughterhouses in Ilam in all seasons between 2017 and 2018. The abomasum content and abomasal mucosa were removed and washed. The collected nematodes were morphologically identified. The genomic DNA was extracted and a 300 bp-fragment-length from internal transcribed spacer 2 ribosomal ribonucleic acid (ITS2-rRNA) gene was amplified. Overall prevalence was 66.70% (160/240). Five species of four genera of nematodes including Marshallagia marshalli (43.70%), Ostertagia circumcincta (15.50%), Parabronema skrjabini (5.00%), M. occidentalis (2.50%), and Haemonchus contortus (0.04%) were identified. Ostertagia circumcincta and H. contortus were found to be different in two nucleotides. There was no nucleotide difference between M. marshalli and M. occidentalis. This study revealed a significant prevalence of parasitic nematodes in sheep abomasum and species diversity of Trichostrongylid nematodes in the region.

In this study, 22 Lactobacillus plantarum strains isolated from Siahmazgi traditional cheese were evaluated using different tests including resistance to low pH (1.50 and 2.50) and bile salt (0.50 and 1.00%), growth kinetic at low pH values and survival under simulated gastric and intestinal fluids. All the strains retained their viability at pH 2.50. However, the survival of all of the isolates was decreased at pH 1.50. Ten out of 22 strains which were able to tolerate low pH were selected for further investigations. All the selected isolates were able to grow at low pH. Strain F2 showed the highest specific growth rate. Five out of 10 isolates showed a significant decrease in bacterial count varied from 2.00 to 7.00 log CFU mL-1 during 3 hr exposure to 0.50% bile salt, while five isolates represented resistance to 0.50% bile during 3 hr. A significant reduction was observed in survival of all of the isolate at 1.00% bile salt concentration. Furthermore, viability of the selected isolates was lowered during 1 hr incubation under gastric conditions, while it remained unchanged within next 2 hr. Although, no significant changes were seen in bacterial count of the selected isolates during 1 hr of exposure to simulated intestinal condition, the survival of the isolates was relatively reduced after 3 hr. In conclusion, five out of 22 examined L. plantarum isolates showed appropriate resistance properties, therefore, could be good candidates for further examinations including functional and safety evaluation supporting their use as probiotics.

Introduction of new inhibitory agents such as peptides, heterocyclic derivatives and nanoparticles (NPs) along with preventive proceedings are effective ways to deal with standard and drug-resistant strains of microorganisms. In this regard, inhibitory activities of some recently synthesized 4-thiazolylpyrazoles, imidazolidine- and tetrahydropyrimidine-2-thiones and magnesium oxide (MgO) NPs alone and in combination with nisin have been assessed against Aspergillus fumigatus. Antimicrobial susceptibility tests were done via broth microdilution, disk diffusion and streak plate methods according to the modified Clinical and Laboratory Standards Institute (CLSI) guidelines. Synergistic effects were also determined as fractional inhibitory concentration (FIC) and fractional fungicidal concentration (FFC) values. Inhibitory potentials of all heterocycles and NPs against A. fumigatus were proved based on inhibition zone diameter (IZD) values in the range of 7.72 - 16.85 mm, minimum inhibitory concentration (MIC) values in the range of 64.00 - 512 µg mL-1 and minimum fungicidal concentration (MFC) values in the range of 256 - 2048 µg mL-1. Tetrahydropyrimidine derivative 3f showed the best inhibitory properties. Inhibitory activity was not significant with nisin. While antifungal effects of major derivatives were improved by combination with it. The results indicated that the combined treatment of heterocycles used in the present study with nisin might be efficient for mold prevention and removal in foodstuffs or other products.

Foot-and-mouth disease (FMD) is endemic in Iraq. The current study can be considered as the first molecular characterization of serotype O in Iraq. The present investigation reported the determination of FMDV serotype O from local farms in Sulaimani districts in 2016 outbreaks. Samples were collected from suspected cattle. The virus was primarily detected with RT-PCR directly from mouth epithelial samples. The direct sequencing and subsequent analysis of amplified PCR products for the VP1 gene indicated the circulation of serotype O of FMD in studied areas. Moreover, phylogenetic analysis revealed that the field isolated serotype O belonged to topotype ME-SA and lineage PanAsia II and clustered with Pakistan and Iran isolates (KU365843 and KY091283) with identity (96.00%, 95.00%) respectively. Furthermore, according to the phylogenic tree, the field isolates had different lineage with the three O/Manisa vaccine strains and Iraq/2000 strains. These findings highlighted the continuous circulation of serotype O of FMD in the region.

Sustained release drug formulations are frequently developed to reduce dosage frequency and to improve outcomes of drug therapy. This study evaluates the pharmacokinetic (PK) parameters of a novel injectable danofloxacin (DANO) formulation in comparison with a conventional product in an animal model. A recently synthesized DANO formulation, prepared by incorporation of DANO-loaded mesoporous silica nanoparticles in liposomes and integration of liposomes in chitosan and β-glycerophosphate solution (lipogel) along with the conventional DANO product were injected subcutaneously (SC) in rabbits. Blood samples were collected at specific time points and DANO concentrations in plasma samples were measured. The PK parameters including maximum concentration (Cmax), time to reach Cmax (Tmax), area under the concentrationversustime curves (AUC), area under the first moment concentration-time curve (AUMC) and mean residence time (MRT) were studied by non-compartmental analyses. The values of MRT (156.00 ± 20.00 hr), AUC (15.30 ± 3.00 µg mL-1 perhr) and Tmax (4.70 ± 1.60 hr) for lipogel formulation were higher than those of the conventional product (8.50 ± 3.60 hr, 3.70 ± 2.00 µg mL-1 per hr and 0.80 ± 0.26 hr, respectively). However, Cmax values for lipogel formulation (0.41 ± 0.15 µg mL-1) were significantly lower than those of the conventional drug product (0.68 ± 0.09 µg mL-1). It was concluded that the novel DANO lipogel effectively slowed down the drug absorption and the incorporation of liposomes in hydrogel could be a useful approach to maintain the therapeutic drug level for a longer period; however, more studies are needed in this field.

In February 2019, a severe respiratory distress with co-infection of infectious laryngotracheitis (ILT) and Newcastle disease accompanied with Salmonella Enteritidis occurred in a broiler flock in the western region of Iran. Clinical signs included paralysis, torticollis, nasal discharge, conjunctivitis, gasping and respiratory rale with high mortality. At necropsy, caseous diphtheritic membrane adherent to the larynx and trachea was observed. Microscopically, syncytial cells formation with dense eosinophilic intranuclear inclusion bodies were main histopathological findings in tracheal tissues. Conventional polymerase chain reaction (PCR) for ICP4 gene amplification as a definitive diagnosis was utilized for the detection of ILT virus nucleic acid in suspected tracheal samples inoculated on to the chorioallantioc membrane of 11-day-old specific pathogen free (SPF) chicken eggs. Tracheal tissues taken from these SPF birds were positive by nested ILT PCR. In conclusion, because of no vaccination policy against ILT in broilers, the most probable scenario is that virus-laden dust or other fomites can be vectors and virus persistence and disease outbreak can be a sequel of wild virus introduction to the farm.

Spondylolisthesis known as kinky back syndrome is a deformity of the free sixth thoracic vertebra including enlargement and rotation resulting in posterior paralysis in broiler chicken. Genotype, feeding regimes and environment are involved in occurrence of this congenital anomaly. Spondylolisthesis may result in or happen simultaneously with two other major axial skeleton pathologies including spondylosis and spondylitis as a result of occurrence of degenerative necrobiotic lesions and inflammation of the vertebrae, respectively. This report deals with a male broiler chicken with posterior paralysis in the second week of rearing. After euthanasia, the specimen was evaluated at macroscopic and microscopic levels. Macroscopic findings showed the unilateral enlargement of body and transverse process of sixth thoracic vertebra and downward rotation of the body. The histopathological studies revealed the focal chondronecrosis and degenerative changes in articular facets and cranial process of the vertebra. In conclusion, a diagnosis of spondylolisthesis followed by spondylosis was made based on macroscopic and histopathological findings that to the authors’ best knowledge, is the first report in a broiler farm in Iran.