فصلنامه دامپزشکی ایران
سال بیستم شماره 83 (تابستان 1403)

One of the methods of protection and slow release of vaccine antigens is the use of biodegradable natural polymers such as chitin in the bacterial biofilm substrate. The bacterium is complex and protected by the polymer substrate in the conditions of the digestive system, and finally, healthier vaccine antigens with better immunogenicity are provided to the intestine mucosal immune system of the fish. In this method, biofilm vaccine of Aeromonas hydrophila bacteria was done by culturing the bacteria in TSB medium (0.225% W/V) along with chitin (0.3% W/V) for 4 days in a shaker incubator. Afterward bacteria were washed with sterile PBS and were inactivated by heat. A number of common carp of 32 ± 11 g were divided into 5 equal treatments and each treatment was repeated three times as follows: the first to fourth treatment with biofilm vaccine, bacterin, media chitin and chitin + TSB respectively, by oral method during 10 days were immunized; the fifth treatment without vaccine administration was considered as the control group. Sampling of fish serum was done on days 0, 20, 40 and 60. Antibacterial antibody level in the samples was evaluated by ELISA test and by monoclonal antibody. The results showed that the antibody level of Aeromonas hydrophila in oral biofilm treatment on day 20 was significant compared to the other treatments, but it was not significant on days of 40 and 60.

Repeated exposure to iron oxide nanoparticles can lead to structural disorders in the testicles and have negative effects on the individual's reproductive function. On the other hand, spirulina algae can reduce these effects by having antioxidant properties and abundant vitamin source. The purpose of this study was to investigate the effects of spirulina algae on tissue changes in the testes of rats induced using iron oxide nanoparticles. After acclimatization, 36 Wistar male rats were randomly divided into the following 6 groups. The first group: control, the second group: 15 mg/kg body weight of iron oxide nanoparticles, the third group: 300 mg/kg body weight of spirulina algae, the fourth group: 15 mg/kg body weight of iron oxide nanoparticles and 300 mg/kg algae body weight. spirulina at the same time, fifth group: 15 mg/kg of body weight of iron oxide nanoparticles for 14 days, then 300 mg/kg of spirulina algae, sixth group: 300 mg/kg of body weight of spirulina algae for 14 days, then 15 mg/kg of body weight of iron oxide nanoparticles. Sampling was done after 60 days and testicular bud sections were stained by hematoxylin-eosin (H & E) after preparation for histological and histometric studies. The study of tissue sections of the testis in rats exposed to iron oxide nanoparticles showed a decrease in the diameter of the spermatogenic tubules and the thickness of the epithelium of the spermatogenic tubules, a decrease in the number of Sertoli cells and the number of Leydig cells in comparison with the control group, while exposure to spirulina algae increased the diameter of the spermatogenic tubules and the thickness of the epithelium of the spermatogenic tubules. Algae treatment in the groups exposed to iron oxide nanoparticles improved the tissue changes in the testis and prevented the decrease in the diameter of the spermatogenic tubes, the thickness of the germinal epithelium, and the number of Leydig and Sertoli cells. The results showed that the oral administration of spirulina in male rats improves the structural disorders in the testis and tissue changes caused by induction by iron oxide nanoparticles.

The current study was conducted to evaluate the effect of pigeon plasma egg yolk (PPEY) and chicken plasma egg yolk (CPEY) compared to combination of PPEY+CPEY in tris-citric acid-fructose diluent to preserve the ram semen quality during liquid-cold storage. Semen samples were collected using the artificial vagina from four Qezel rams twice a week. Then, in case if the samples met the criteria, they were pooled and used for the experiment. Tris-citric acid-fructose based extenders were prepared using PPEY (28%, v/v), CPEY (28%), and the combinations of PPEY (7, 14 and 21%) + CPEY (21, 14 and 7 %), and used for the experiment. Following dilution of samples with extenders, total and forward progressive motility of spermatozoa (evaluated by computer assisted sperm analysis), viability and plasma membrane integrity was assessed at 0, 24, 48 and 72 of after cooling. Furthermore, amounts of malondialdehyde (MDA), as an oxidative indicator, were measured at mentioned time points. Results revealed that forward progressive motility (at 24, 48 and 72 h) and viability (at 72 h) were higher in PPEY (28%) group compared to all combination groups. Moreover, membrane integrity (at 24, 48 and 72 h) was greater in PPEY (28%) relative to combination groups containing 14 and 21% CPEY. Amounts of MDA did not differ among treated groups. In conclusion, combinations of PPEY+CPEY was not effective as PPEY (28%) and CPEY (28%) alone to preservation of ram semen during liquid-cold storage.

The goal of this study was to detect and evaluate the presence of primordial germ cells in pheasant ovarian tissue at various embryonic ages. These are the only growing cells in the embryo capable of transmitting genetic information to the following generation via gametogenesis. Birds are quite useful for research in this area because of their growth features. On the other hand, the pheasant is one of the birds with very beneficial features, but this rare species is at risk of extinction. In this study, 28 fertilized pheasant eggs were used, and the gonads of female embryos were differentiated based on their asymmetry and retrieved between embryonic days 9 and 24. Tissue sections were prepared as serial sections and the identification of primordial germ cells was examined using hematoxylin-eosin and PAS staining, as well as semithin slice preparation and toluidine blue staining. The number of primordial germ cells, cell volume, and nucleus volume of primordial germ cells were then estimated using stereological techniques. Early germ cells were recognized in paraffin and resin sections from embryonic days 9 to 14 because of their unique properties, which comprised a big nucleus and little cytoplasm. Furthermore, the response of these cells to PAS staining was negative at all ages. According to the stereological results, the number of primordial germ cells was decreased at more advanced ages of development, so that it was not seen at 18 day embryo. Whereas, the volume of the primordial germ cells and nucleus of these cells have been increased with the increasing of the age. The findings of this study revealed that primordial germ cells in the ovarian tissue of pheasant embryos at all examined ages responded negatively to the PAS staining, whereas routine hematoxylin and eosin staining and the use of semithin sections are suitable for microscopic detection of these cells. Morphologically, this cell in pheasant was similar to other birds, whereas in terms of histochemistry, the reaction of these cells was negative to PAS staining which differs from some species of birds.

Honey bees play an important role in the survival and pollination of many plants. One of the most important of pests that attack honey bees is Varroa destructor mite. This mite is an ectoparasite of honeybees and feeds mainly on larvae and pupae. In this research, Apis mellifera meda workers were infected at the pupal stage to different mite densities and then the biological and morphological characteristics of the emerged adult bees were investigated. For this purpose, female mites were collected from infected hives and placed in each prepupal cell according to the desired mite density (0, 1, 2 or 3 mites). After adult bees emerged, nutritional traits were checked every 3 days for 12 days. Then these adult bees were dissected to measure the area of ​​the hypopharyngeal acinus. The biological and morphological results showed that the increase mite density had no effect on the duration of the pupation period of bees, but it significantly decreased the weight, body length, area of the forewings, hind wings and hind tibia of bees. Adult bees that had three mites during their pupation had the highest mortality rate and for this reason, it was not possible to examine their nutritional characteristics. The nutritional characteristics results showed that increasing the mite density caused a decrease in the consumption of syrup and water by the adult bees, but the consumption of pollen did not change. The food consumption index of bees without mite and bees with two mites during the pupation period was higher than bees with one mite, but the growth rate and the area of the acini of the treatments did not show any difference. The results of this study are expected to provide a basis for biological control of this mite by providing further information on how it affects honeybees.

Mineral oils are the most common adjuvants that are used in the production of poultry killed vaccine based on water-in-oil emulsion (W/O) to create high and long term antibody responses to vaccine antigens. The vaccine structural specifications, including particle size and formulation on stability and immunogenicity, have needed more clarification for vaccine producers so that they can produce their products based on highly efficient methods. This study assessed the effect of particle size on the stability and immunogenicity of a W/O vaccine. Different W/O emulsions were formulated by three different homogenization times by means of inactivated Avian Influenza Virus subtype H9N2 (AI H9N2) and Newcastle Disease Virus (V4) antigens and MontanideTM ISA 70 adjuvant. The three killed AI+ND vaccines were formulated and 2 commercial vaccines were evaluated for magnitude and duration of specific antibody response in SPF and broiler chickens. The vaccine dose for each bird was considered 0/2 ml. On days 7, 14 and 21 post vaccination, antibody titers of serum were evaluated by Hemagglutination Inhibition (HI) test. The serum antibody titers (ND) of group B and C on days 7 and 21 were higher than the other groups (P<0.05), but the difference between the other groups was not significant on the same day. The serum antibody titers (ND) of broiler chickens in group B and C on day 7, 21 and also group E on day 21 were significantly higher than other groups. The serum antibody titers (AI) of SPF chickens on day 21, in group C, had significant difference with other groups. The antibody titers (AI) on days 7 and 14 showed no significant difference in all groups except group D. The immunogenicity of emulsions was higher in more homogenization time.  In all, these results indicate that increases in homogenization time create a smaller size of droplets and are effective for the rapid onset of antibody production. The small size of droplets not only creates a fast and good response, but also promotes immunogenicity both in broilers and SPF chickens.

The main objective of this study was to compare the accuracy of renal volume measured with ultrasonography to real volume in ten native dogs that were candidates for euthanasia due to severe injuries sustained in traffic accidents. Furthermore, correlations of renal dimensions in ultrasonography, such as length, width, and height, were determined with real ones. Initially, the dimensions of the right and left kidneys were determined using ultrasonography. Renal volume was measured using two different methods. The first method consisted of choosing automatic volume measurement in an ultrasonography unit. The volume formula of a prolonged ellipsoid (length × width × height × 0.523) was used in the second method. The real renal dimensions (golden standard) were measured during celiotomy and after necropsy, and the true volume of kidneys was calculated using water displacement. Linear regression analysis was used to determine correlations between renal dimensions and volume in ultrasonography with real ones. Results showed that renal dimensions and volume measured in ultrasonography were lower than actual measurements. There were positive and statistically significant correlations between renal volume in ultrasonography and actual renal volume. However, the correlation coefficient (r) of volume determined by the formula was greater than that of the automatic volume method, and its mean was closer to that of the real volume mean. There were also positive and significant correlations between renal dimensions measured with ultrasonography and those measured in real measurement. No significant difference was found between renal dimensions measured during celiotomy and after necropsy. These findings indicate that ultrasonographic measurements underestimated renal dimensions and volume, and the formula method provided more accurate results than the automatic volume method.

The Hemiscorpius lepturus scorpion is known as the most dangerous scorpion in the south and southwest regions of Iran. This scorpion can cause illness and even death in humans and animals. The purpose of this study was to investigate the metabolic and endocrine effects of different fractions of Hemiscorpius lepturus venom. For this purpose, scorpion venom was extracted by electric shock method and after separating the solution phase and activation of the venom by freeze drying method and with the help of column chromatography by Sephadex G-250 gel, 6 fractions were separated based on optical absorption at 280 nm. Then 72 male Wistar rats were divided into 8 equal groups including control (NS), crude venom (1000 µg/kg), fractions I (120 µg/kg), II (430 µg/kg), III (80µg/kg), IV (180 µg/kg), V (60 µg/kg) and VI (130 µg/kg) divided and according to the above amounts, respectively, with normal saline, crude venom and different fractions through intraperitoneal injection. After 1, 3, 24 and 72 hours, the concentration of glucose and the insulin, glucagon and cortisol were measured by elisa with specific rat kits. The results showed that the injection of crude venom and all fractions caused a significant increase in glucose compared to the control group. Also, a significant increase of cortisol and glucagon was observed after injection in crude venom and fractions II and VI. Also, the average insulin in the groups receiving crude venom and fractions II and VI showed a significant decrease. These findings showed that the injection of different fractions of Hemiscorpius lepturus scorpion venom can affect the metabolic process and cause changes in the metabolic processes by affecting the glucose regulating hormones. Identifying these effects, considering the wide effects of these hormones in metabolism, as well as the changes caused by the venom fractions, can help to identify the toxicity mechanisms as accurately as possible and also to adopt specific treatment methods in cases of scorpion stings. be helpful and useful.

Reactive oxygen species (ROS) and the oxidative damage are the main factors involved in tissue damage caused by arsenic trioxide. In the present study, we investigated the protective effects of royal jelly on the parameters of oxidative stress and histopathological damage to lung tissue due to arsenic trioxide toxicity. Forty male Wistar albino rats received arsenic trioxide and royal jelly orally for 30 days and were randomly divided into eight groups of five; The first group received normal saline at a dose of 1 mg /kg, the second group received royal jelly at a dose of 150 mg /kg, the third group received arsenic trioxide at a dose of 1 mg / kg, the fourth group received arsenic trioxide (1 mg /kg) with Royal jelly at a dose of 150 mg /kg, the fifth group received arsenic trioxide (1 mg /kg) with royal jelly at a dose of 100 mg /kg, the sixth group received arsenic trioxide (1 mg /kg) with royal jelly at a dose of 50 mg /kg, the seventh group received arsenic trioxide (1 mg /kg) with vitamin E at a dose of 100 mg /kg, and the eighth group received arsenic trioxide (1 mg /kg) with vitamin E and royal jelly (both) at a dose of mg/ kg 100. Oxidative stress parameters including malondialdehyde (MDA), glutathione peroxidase (GPx), superoxide dismutase (SOD) and catalase (CAT) were evaluated. Damage to lung tissue was also assessed by hematoxylin-eosin (H&E) tissue staining. The results of the present study revealed that arsenic trioxide increased the level of MDA and decreased the levels of GPx, SOD and CAT. Royal jelly treatment decreased MDA levels and increased GPx, SOD and CAT concentrations in lung tissue. The severity of lung tissue damage was also reduced. In general, Royal Jelly reduced oxidative stress and lung tissue damage.

The set of non-specific antioxidant activities is called Total Antioxidant Capacity (TAC). The aim was to determine the serum TAC level following selenium and Vit.E supplementation in lambs. Thirty two male Macuei lambs under 2 months of age and average weight of 10 kg were selected. Lambs were classified into 8 groups including control, Vit.E, Se nanoparticles (NanoSe), Oral sodium selenite (NaSe), NaSe Injection, NanoSe/Vit.E, Oral NaSe/Vit.E and Se/Vit.E (E/Se) injection and studied for 90 days. Lambs were bled 6 times around 5 ml at days 1, 7, 14, 30, 60 and 90 and sera were used to assess TAC consentration. In all, the lowest TACs in lambs was observed in the NanoSe group (1.17 μmol/l) and the highest was in the injected E/Se group (1.32 μmol/l) on day 60, but the differences among 8 groups during the 90 days were not significant. During the 90 days of investigation the most stable and lowest changes in TAC concentration was observed in the Vit.E group and the most unstable changes were revealed in the NanoSe and injected NanoSe/Vit.E groups. The mean TAC concentrations on days 14 and 60 were significant among groups, which was mainly predominant in the injected sodium selenite and injected selenium/vitamin E groups, respectively. Comparison of TAC concentration among sampling times revealed the difference in NanoSe and NanoSe/vitamin E in which the lowest concentration belonged to NanoSe. The results of the interaction effect of sampling time and groups on TAC concentration showed a significant time difference, especially on days 14 and 60, with no significant difference between groups. TAC concentration in the first day was correlated with day 60 in the control group, day 7 and 14 in NanoSe/Vit.E, group and day 60 in injected E/Se. In conclusion, selenium and Vit.E supplements had no effect on TAC levels, whereas TAC levels were time-dependent. Therefore, the activity of nonspecific antioxidants (TACs) is not affected by specific antioxidants such as selenium and Vit.E and changes over time.

Stanozolol and nandrolone decanoate hormones are mainly used in dogs and horses to strengthen muscle growth, increase of body weight, treatment of anemia and stimulation of appetite; nevertheless, some side effects have been reported, including testicular atrophy, and decrease of spermatozoid count in dogs. The aim of the present study is to evaluate the histopathological changes of testicular and epididymal tissues in dogs treated with stanozolol and nandrolone decanoate. The present survey was performed on sixtheen male dogs. The studied animals were divided into two equal groups of eight. Stanozolol was administered to dogs in group A with dosage of 50 mg per dog intramuscularly for 6 continuous weeks. Group B was similar to group A, but instead of stanozolol, nandrolone decanoate was injected with dosage of 1 mg/kg. To evaluate the possible side effects on the testicles, one of them was surgically removed before the investigation and the second testicle was removed on days 28 (four dogs in each group) and 42 (the remaining four of each group). Before the testes were placed in 10% formalin buffer, sperm quality in the epididymis was evaluated for indicators such as motility, morphology and number in times zero, 28 and 42 days. The results showed that destructive effects were obtained such as hyperplasia of interstitial Leydig cells, vacuolation, the presence of multinucleated cells and different stages of spermatogonia in testicular tissue; also, the destruction of duct covering cells and the detachment of cilia in the epididymal epithelium was observed. These changes were significantly greater in the stanozolol group than nandrolone decanoate. The quantitative changes were more pronounced in the stanozolol group and the qualitative changes in the nanderlone decanoate group. Although both drugs had side effects on testicular and epididymal tissues; the side effects of stanozolol were greater than nandrolone decanoate; therefore, it is recommended minimum dose of nandrolone decanate for therapeutic purposes.

Ornithobacterium rhinotracheal (ORT) is a Gram-negative pathogen that is one of the causes of respiratory diseases in birds. Co-infection of Ornithobacterium rhinotracheal with other pathogens can cause serious health problems in bird species and also lead to financial losses in the poultry industry. Serological methods are among the most important techniques for rapid infection diagnosis, farm monitoring, infection control and prevention. The purpose of this study was to design an ELISA test to assess the prevalence of ORT and track the amount of ORT-specific antibody in serum collected from turkeys.  After immunizing three pieces of turkey and obtaining the immunization serum, antibody purification was performed using ion exchange chromatography and affinity chromatography methods. The purified antibodies were then used in a homemade ELISA reaction as a standard sample to determine the quantity of specific antibodies against ORT. A total of 244 serum samples were collected from turkeys slaughtered in the poultry slaughterhouse of Khuzestan province. ELISA testing revealed the significant presence of antibodies against ORT in 141 (57.78%) of the samples. The average concentration of antibodies in positive samples was 467.72 ± 124.128 micrograms per milliliter of serum, while in negative samples it was 262.97 ± 37.263 micrograms per milliliter. This study revealed the prevalence of ORT infections in Khuzestan province. Therefore, due to the lack of a reliable vaccine against O. rhinotracheal, it is necessary to implement good disease management and biosecurity measures in order to effectively control the disease.