Veterinary Research & Biological Products
Year:2020 | Volume:33 | Issue:1 (126)|Papers Count:15

Detection and identification of influenza A viruses hemagglutinin is done with serological or molecular techniques and the neuraminidase (NA) subtyping is also necessary in purpose of epidemiological studies specially in H5 or H7 (highly pathogenic/ low pathogenic) outbreaks and/or endemic areas. Subtype Identification of NA usually is performed by molecular tests such as RT-PCR. Since 1971, WHO has suggested that the surface glycoprotein neuraminidase of avian influenza viruses could be divided in different subtypes serologically by Neuraminidase Inhibition test (NI). In reference or specialized laboratories like OIE licensed labs, NI test is running routinely. Considering the importance of this valuable test, an experimental study was conducted to develop the classic NI test according to OIE protocol. RT-PCR test with subtype-specific primers was used as a control for evaluation of test accuracy. According to our results, NI test can detect and distinguish various NA subtypes in serum or viral samples. In conclusion, the NI test is a practical tool in diagnostic and research trials, especially in differentiation of vaccinated from unvaccinated birds (DIVA) in endemic areas with vaccination strategy or in validated reference laboratories such as Razi Vaccine and Serum Research Institute.

Leptospira interrogans is the cause of leptospirosis, a common disease among humans and animals. One of the most important recommendations of international associations concerned with laboratory animals, is the use of defined animals. Based on the recommendation of the FELASA, the diagnostic monitoring is required to evaluate the contamination of animals to different microbial agents. In this study, the prevalence of this bacteria was investigated in a colony of NIH strain laboratory mice of laboratory animal breeding unit at the Razi Institute in 1396. 74 adult mice (in the range of 30-35g and age 6-8 weeks) were randomly selected from both sexes in breeding colony and 6 wild mice were selected by means of live traps from the outside of facility. After ethical euthanasia and autopsy, samples were taken from their kidney tissues and after preparation, the PCR method was performed using Leptospira interrogans specific primers. Designing and preparing the positive control was carried out using plasmid (pGEM-T easy vector) transformation containing the pathogenic of Leptospira interrogans gene (LipL32) in E. coli for gene amplification and extraction. No positive cases were observed from the examined samples. According to FELASA instructions, it can be concluded that with 99. 9% confidence, the Leptospira interrogans infection in breeding salons of Razi Institute NIH mice was discarded.

Staphylococcus aureus considered as one of the commonest agents of bacterial food poisoning which may transfer to human from animals and animal-originated foods. Therefore, the aim of this study were isolation of the S. aureus from native sheep nose of Sistan region; evaluation of antibiotic resistance of isolated strains treated with common antibiotics; and studying the antimicrobial effects of ethanol extracts of Withania somnifera and Rosmarinus officinalis on isolated strains. A total of 15 strains of S. aureus were isolated through biochemical methods. Ethanol extracts of W. somnifera and R. officinalis obtained using Rotary apparatus and distillation in vacuum methodology. Susceptibility of the isolated strains to common antibiotics was determined via Kirby-Bauer disk diffusion standard test. Through dilution in well method susceptibility of the isolated strains were determined in treatment with ethanol extracts of W. somnifera and R. officinalis. Among evaluated antibiotics, the studied strains were not influenced by Oxacillin but Gentamicin controlled the bacterial growth completely. In comparison with R. officinalis, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of W. somnifera arises in a lower amount to inhibit the growth of the bacteria. While due to susceptibility of the bacteria to the common antibiotics, treatment of the bacteria with antibiotics resulted in rapid decreasing in the bacterial population, however, due to probable antibiotic resistance, it is recommended that antibiotics would be replaced with plant extracts. Comparison between two treated extracts in the current study have shown that ethanol extract of W. somnifera could be introduced as a suitable replacement with common antibiotics.

In most of parasitic helminths infection in gastrointestinal (GI) and respiratory systems, mix infection plays important role. This study was aimed to determine parasitic helminths fauna of GI and respiratory systems using fecal examination and culture. A total of 140 fresh fecal samples were randomly collected from the rectum of examined sheep in different parts of Gilan Province in 2017-2018. A portion of each sample subjected for floatation and modified McMaster techniques. The intensity of infection estimated in term of eggs per gram of feces (EPG). Helminths identified based on eggs’ morphometric and morphologic characteristics. Third stage larvae (L3) and first stage larvae (L1) were collected using Baermann method and identified. Overall prevalence was 52. 1% (73/140) with intensity of 196. 62± 9. 94. The higest prevalence was found in Siahkal region (7. 9%) in spring (39. 3%). Helminths diversity was from trematodes (9. 3%) (Fasciola hepatica (4. 3%), Dicrocoelium dendriticum (5%)), cestodes (10%), and nematodes (52. 1%). All infected sheep had mixed helminths’ eggs infections with two (15%) and three (2. 9%) genus. Identified L3 of nematodes in GI was nine genus including of Ostertagia (4. 3%, 2 species), Haemonchus sp. (1. 4%), Trichostrongylus sp. (1. 4%), Bunostomum sp. (2. 1%), Strongyloides sp. (0. 7%), Cooperia sp. (1. 4%), Nematodirus sp. (2. 1%), Chabertia sp. (1. 4%) and Oesophagostomum sp. (2. 9%). Identified L1 of lungworm (11. 4%) was four species including of Dictyocaulus filaria (7. 1%), Protostrongylus rufescens (2. 1%), Cystocaulus ocreatus (1. 4%) and Muellerius capillaris (0. 7%). From the results, it was concluded that helminths infection of GI tract and respiratory system was prevalent in examined sheep of the region.

In this study, the anthelmintic effects of the medicinal herb, Nigella Sativa L., was screened in vitro against the infective larvae of Parascaris equorum. Follow the collection of Parascaris worms, the egg content were depleted and hatched using sodium hypochlorite solution. The recovered larvae of the parasite were exposed to six concentrations (6. 25, 12. 5, 25, 50, 100 and 200 mg/mL) of the Nigella extract and then examined for the viability at 0, 10, 20, 30, 40, 50 and 60 minutes after the challenge. The results revealed that the concentration of 6. 25 mg/mL had not any significant anthelmintic effects on P. equorum larvae. In contrast, the statistics indicated that all the higher concentrations of the Nigella extract remarkably reduced the viability of larva. These results were obvious during the first ten minutes of the experiment. With increase in time of exposure, the mortality effect of the extract elevated significantly. In addition, all larvae were inactivated after 40, 20 and 10 minutes after exposure at the concentrations of 50, 100 and 200 mg/ml, respectively. These results confirmed that this herbal extract possess good antiparasitic effects against P. equorum infections.

Honey bee ectoparasites infestations may affect the insect and cause the population reduction and economic damage in apiculture system. The present study was aimed to determine the Prevalence of Varroa and association of Colony Collapse Disorder (CCD) in apiaries in Urmia suburb, Iran, over one year (2017-2018). A total of 840 beehives of 120 apiaries in Urmia suburb were randomly sampled. Of those, 301 bees (35. 83%) and 190 bee larvae (18. 33%) were infested with Varroa. The highest infestation was found in bees and bee larvae in summer. 93 out of 840 apiaries had CCD signs. 61 out of 93 apiaries (65. 59%) with CCD were infested to Varroa. There was a significant relation between CCD and Varroa (p <0/05). From the results of this work, it was concluded that Varroa mites are a prevalent ectoparasite in apiaries of Urmia suburb which may have an important role in the incidence of CCD.

Unfortunately, using common antibiotics to treat animal infection is increasing in which leads to undesirable side effects such as antibiotic residuals in animal products and occurrence bacterial resistance to antibiotics. Therefore, researchers are searching for an alternative to traditional antibiotics with fewer side effects like herbal essential oils and antimicrobial peptides (AMPs) such as oregano and cinnamon essential oils as well as recombinant thanatin peptide. Minimum Inhibitory Concentrations (MIC) was used based on a microbroth dilution method by 96-well microtiter with four replicates with oregano and cinnamon essential oils as well as thanatin peptide. After MIC, bacteria species were cultured and minimum bactericidal concentration (MBC) was determined as that showing no growth. The results of MIC assay showed a strong activity against Salmonella enteritidis bacteria with the value of MIC equal to 6 μ g/ml by thanatin. Moreover, the lowest amount of MBC was observed on E. coli0157 and Salmonella enteritidis approximately 25 μ g/ml. Thanatin is one of the member of cationic AMPs that have received a great attention regarding growth inhibitory against wide range of microorganisms including Gram-negative and Gram-positive bacteria, as well as several fungi species. Thanatin showed a strong growth inhibitory against bacteria in compare with oregano and cinnamon essential oils. Thus, developing new antibiotics such as thanatin would be a great replacement for common antibiotics and may lead to a great progress in eliminating livestock pathogens.

Today, salmonella is one of the most important causes of food disease in the world. The purpose of this study was to evaluate the effect washing stages of carcasses and chiller in contamination rate of carcasses to Salmonella in industrial Slaughterhouses. In this study five carcasses were randomly sampled for each stage of the six stages of slaughter of poultry so from every slaughterhouse 30 carcass samples and total 180 samples were taken of 6 slaughterhouses which are active in the Khuzestan Province. After rinsing carcasses with peptone water medium of each carcass was prepared 100 ml sample (carcass rinses solation) which were sent to the laboratory for microbiological tests. In the research was determined Salmonella contamination In 65% (117 samples), tested carcasses were positive and in 35% (63 samples) negative. Salmonella contamination was determined in samples after Scalding 56. 6%, after plucking 66. 6%, before evisceration 60%, after evisceration 70%, before the chiller 63. 3% and after the chiller 73. 3%. The results showed significant effect washing carcasses in reduce contamination to Salmonella and the significant effect of the chiller (immersion) stage on increasing the carcass contamination with Salmonella.

To investigate the effects of dietary zinc and betaine (Bet) substitution to methionine on performance, egg quality and blood parameters in laying hens under heat stress, an experiment with 288 Hy-line W-36 leghorn (at 50 to 62 weeks old) in complete randomized design (CRD) with a 3×2×2 factorial arrangement of treatments with twelve treatments, 3 replicates and 8 hens in each replicate was performed. There were 6 dietary treatments: three doses of Bet (0, 13 and 26%) substitute to methionine combined with three levels of zinc (50 and 100 mg/kg) supplemented to the basal diets that were combined with two environmental conditions (thermoneutral (23° C constant) and heat stress ( 23. 9 to 35° C cycling) conditions). The results indicated that performance parameters of laying hens and egg quality were all significantly reduced by heat stress (HS) (P<0. 05). Substitution of methionine with Bet led to significantly increase of egg production (P<0. 05); whereas it had no effect on egg weight and egg quality. Supplementation of the basal diets with zinc significantly increased laying performance of hens and egg quality (P<0. 05). Moreover, the results of this experiment indicated that, there were significant effect of Betaine on blood cholesterol and triglyceride, (p<0. 05), but no significant effect on glucose in broiler chickens. Moreover, results demonstrated that Zinc supplementation decreased cholesterol, triglyceride and glucose. The present study showed that dietary supplemental Bet and zinc at the level of 13% and 100 mg/kg resulted in an improving egg quality and performance of laying hens under heat stress.

The chromaffin cells play a role in maintenance of blood pressure, carbohydrate metabolism and blood flow to vital organs during stress. Distribution of these cells is different in domestic mammals. Dispersion of these cells were studied in 40 one humped camels from 6 to 10 years old, after sectioning and staining with Hematoxylin-Eosin, Chromaffin and Mason’ s trichrome. The gland is enclose within a capsule of dense collagenous connective tissue and many trabeculae separated from it and penetrated into the cortex. Medulla separated from cortex by a layer of dense collagenous connective tissue. Zona glomerolosa, zona fasiculata and zona reticularis are present in cortex and they constitute about 15%, 55% and 27% by volume of cortex respectively. The zona reticularis is greater in male camels compared with female camels, but there is no significant difference between them. Blood vessels that are surround with dense collagenous connective tissue passed through the cortex and all three layers of the cortex are present around them. Adrenal nodules that enclosed with dense connective tissue are present between cortex and medulla and contained all three layers of the cortex. Medulla extends to the capsule in some areas and cortex is absent in these subcapsular areas. Sometimes the adjacent cortex of these areas only contains zona reticularis. Arrangement of the chromaffin cells has a random pattern in females, whereas they are in two separate layers in males. Epinephrine producing cells are located in outer layer and norepinephrine producing cells in the inner layer of medulla.

Introduction: For fertilization process, sperm cell-surface protein called "IZUMO” requiredas a ligand for IZUMO1R/ JUNO and CD9 receptor on egg. The IZUMO1: IZUMO1R/JUNO interaction is a crucial adhesion step between sperm and oocytein plasma membrane binding and fusion, but also is not sufficient for cell fusion and others receptors are play roles in this interaction. Here, we found G protein-coupled receptor 15 (GPR15) as another partner of sperm-egg interaction which is located on egg. Methods: Initially, system biology methods applied for known and predicted protein-protein network interactions of IZUMO1. Also, primary structure of IZUMO1 and its IZUMO1R: JUNO receptors retrieved from reference databases, and their 3-dimensional structure (3D) were modeled by both Threading and homology modeling. Then, modeled structure were energetically minimized and validated by Rammapageplots and eventually prepared for molecular docking (hydrogen and atomic charges) by Chimera UCSF 1. 10. Also, the docking studies were performed by HEX version 8. Finally, binding energy, pose of interactions, RMSD, hydrogen and electrostatics bonds were analyzed. Results: System biology analysis showed that beside IZUMO1R/JUNO and CD9, GPR15 maybe is another functional partner for IZUMO1 by score of 0. 705 (text mining approach). Also, Rammachandran plot of modeled structures represent high quality of modeling procedure and so modeled structure used for docking. Molecular docking analysis showed GPR15 could interact with IZUMO by SER176, THR91, ARG236 and SER240, ASN239and ASN151hydrogens. The strongest hydrogen bond is between thoronine 91 (THR91) and serine 240 (SER240). On the other hand, amino acids Threonine 91 and ASN239 are the most important amino acids for binding to hydrogen bonding two proteins to each other and react simultaneously with two amino acids. Also, 21 amino acids from IZUMO1 and GRP15 play a role in the hydrophobic bond between two proteins. Discussion: The results of this bioinformatics study can help to find out more about fertility and Sperm fertilization. Further studies on the role of GPR15 in the binding of sperm and oocyte should also be made.

In this study, the effects of broccoli and mulberry leaf on growth performance and intestinal villi morphology in broiler chickens were evaluated. A total of 216 one-day old broiler chickens were randomly divided into 6 groups with 3 replicates of 12 birds. Group 1 received basal diet with standard ration of broiler chickens. Group 2 was fed with a diet containing 500 mg/kg of vitamin C. Groups 3 and 4 received diets containing 0. 5 % and 1 % of broccoli powder, and groups 5 and 6 received diets containing 0. 05 % and 0. 1 % of mulberry leaf ethanoic extract respectively. Variables related to growth performance and intestinal villi morphology were evaluated at age 35. Growth performance increased in experimental treatments in comparison to control, but did not show statistically significant difference. The results of intestinal morphology indicated that in the duodenum, all parameters were higher in experimental treatments than control (P<0. 05). In the jejunum, villi height was higher in all treatments exception to mulberry leaf 0. 05% and absorption surface of the groups vitamin C, broccoli 1% and mulberry leaf 0. 1% was more than control (p<0. 05). In the ileum, vitamin C in all parameters and mulberry leaf 0. 1% had more villus width and lamina propria depth than control (P<0. 05). The present study showed that the addition of broccoli powder and mulberry leaf ethanoic extract to broiler chickens diet improved the intestinal villi morphology in terms of height and absorption surface and could increase the food absorption, so relatively improve growth performance.

Epilepsy is one of the chronic diseases of the central nervous system. Despite the many drugs used to treat it, their effectiveness is questionable. So the purpose of this study was to investigate the stimulatory and protective effect of ketamine on experimental-epileptic activity induced by intraperitoneal injection of pentylenetetrazole in adult male rats. In this study, 15 male rats (200-250 gr) were used and randomly divided to control groups (n=5) that received normal saline and two treatment groups that received ketamine 20 mg/kg (n=5) and the second group that received ketamine 50 mg/kg (n=5) intraperitoneally. After anesthesia with ketamine-xylasein combination and animal skull surgery, the intradermal electrodes were placed on the striatum pyramidal layer of CA1 region of the hippocampus and epileptiform activity was induced by intraperitoneal injection of pentylenetetrazole and was evaluated in terms of the number of spikes per unit of time and amplitude of them by eTrace software. The results of this study showed that ketamine intraperitoneal doses of 20 and 50 mg/kg had no significant effect on the epileptic activity induced by pentylenetetrazole. Intraperitoneal injection of ketamine 20 and 50 mg/kg has no significant effect on pentylenetrazole induced seizure activity and has no stimulatory and inhibitory effects in the desired activities in that doses.

This study was conducted to examine the effect of Zipmet on healing of corneal ulcer due to burning in rabbit. In this study, 20 adult healthy male NewZealand rabbits with an average weight of 2. 5-3 kg were used. After general anesthesia, corneal ulcer in the left eye of the all animals was induced. The left eye of each rabbit was stained with 2 drops of tetraquine 0. 05%. After 5 minutes, a circular filter paper of 6 mm in diameter, impregnated with 30 μ L of a Sodium hydroxide 1 was placed in the cornea center for 60 seconds. After returning from anesthesia, the rabbits were randomly divided into 4 groups of 5 numbers. Then, in the first, second and third groups, Zipmet at 50, 100 and 200 mg / kg were gavaged, while the fourth group was gavaged with water for 21 days. After 21 days, corneal samples were taken and evaluated histopathologically. According to the histopathological findings, vascularization, inflammation and matrix edema of cornea significantly marked while the epithelium of cornea increased in metformin 200 mg / kg-treated rabbits compared to control group (P<0. 05). Also, administration of Zipmet significantly improved the natural structure of cornea collagen fibers (P<0. 05). Based on the results, it seems that Zipmet at dose of 200 mg / kg has useful effect on the healing process in rabbit’ s corneal ulcer.

Snakes are one of the most useful animals in the world for the production of medicines, sera and vaccines that are made from the venom. Venomous snakes have many therapeutic applications and are very useful in biochemical research. Snake venom is an oily liquid, white to yellow with a slightly acidic pH and is a complex mixture of polypeptides, toxic proteins, enzymes, pharmaceuticals and non-protein substances. Venom of all snakes is not the same, although it is believed that venom of snakes originated from common biological ancestors. The aim of this study was to determine the activity of protease enzyme in venom of horned snake from different regions. In this study, the venom of the horned snake from three regions of Semnan, Kerman and Khuzestan was investigated. At first, the venom protein concentration of this snake was determined using Lowry and Bradford methods, which was reported to be 950 mg / ml (95%). In the next step, the number of peptides and molecular weight of these proteins were determined by vertical electrophoresis, which were performed in both reducing and non-reducing methods. And the molecular weight range of 15 to 190 kDa was estimated to have relative similarities in Khuzestan, Kerman and Semnan. Native electrophoresis and casein substrate electrophoresis confirmed the proteolytic activity of ruminant venom and quantitative determination of venom proteolytic activity was determined by BAPNA dilution and casein digestion 11. 56 U/ml.