Archives of Razi Institute
Volume:76 Issue: 2, Jun 2021

The first Attenuated rubella vaccine was developed by Parkman and Meyer in 1966. Ten years later in the 1975s, the rubella vaccine was developed in Razi Vaccine and serum research institute) RVSRI). In 1977, the rubella TAKAHASHI vaccine successfully passed the clinical trial and was initially used voluntarily only in the private sector. Since 1987, the administration of rubella as MMR (Measles/AIK-C; Rubella/TAKAHASHI; Mumps/HOSHINO) strain vaccine has been included in the immunization program in Iran. This review article focused on the development and production of the rubella TAKAHASHI/HDC vaccine in RVSRI. The herd immunity and rubella cases were investigated in the pre- and post-vaccine era. The effectiveness and proper coverage of the rubella vaccine led to the elimination of rubella from Iran in 2019. The current study aimed to assess local rubella vaccine manufacturing and its consequences on rubella elimination from Iran, using various search engines. A complete search was carried out in medical databases, including PubMed, Scopus, Web of Science, Scientific Information Database, IranMedex, Magiran, and Google Scholar. Within 1972-1975, Rubella TAKAHASHI/HDC vaccine was developed by RVSRI and successfully passed clinical trial in 1977. Over the four last decades (1980-2020), more than 40 million infants, young, and adults were vaccinated by million doses of local Rubella, measles-rubella (MR) or measles-mumps-rubella (MMR) vaccine in Iran. In 1972, the pre-vaccine era, the overall sensitivity to rubella infection was 69% in one-year-old Iranian children and 23% in childbearing women. The use of a safe, inexpensive, and effective vaccine increased herd immunity to 95% (85%-99%) in our country. During the last two decades, we have witnessed a 91% decline in the confirmed rubella cases, from 1124 in 2000 to 33 cases in 2018. The current article presented the process of vaccine development, tracked it through more than four decades, and discussed disease status before and after the rubella vaccine era, as well as the history of its elimination from Iran. The effectiveness of the local Razi Rubella vaccine resulted in a significant increase in seroprevalence in Iran. Expanded vaccination against rubella, usually with measles, has led to the elimination of Rubella from Iran as confirmed by World Health Organization in 2019.

Authentication of animal cell lines in cell banks is one of the most important programs regulated during cell culture and storage. This operation provides a thorough and beneficial document which can be advantageous for the functional use of animal cell lines. Therefore, various procedures are used to prevent misidentified cells, cross-contamination to other cell lines, and mislabeling errors leading to incorrect assessment. These contaminants can result in major financial disadvantages. One of the practical methods in this field is a molecular procedure which can demonstrate more accurate results. In the present study, the BHK-21 (C5) was characterized, and it was tried to determine the identity of BHK-21 (C5) as a continuous cell line by Polymerase chain reaction (PCR) molecular procedure in Iran. The cytochrome c oxidase I (CO1) gene was selected as a prevalent DNA fragment for the authentication of the BHK-21 (C5) cell line, along with six cell lines, including Chinese hamster ovary,  Lamb kidney, Razi Bovine Kidney, Medical Research Council cell strain 5, Monkey Green Kidney, and Goat Lymphocyte. After amplification, PCR products were analyzed by agarose gel electrophoresis to ensure their accuracy. The results of characterization were indicated, cell viability was estimated to be about 92%, and a uniform cell culture was obtained. The doubling time and µ ratio equivalent were obtained at 20.5 h and 0.03, respectively. Sterility tests revealed that the cell seed was free of bacterial, mycoplasma, and mycobacterial infections. The results of molecular identification revealed that the identification of this cell line was approved and can be used in studies, diagnosis, production, and quality control of biological products.

Brucellosis is recognized as a zoonotic disease with high morbidity in the absence of treatment. The primary diagnosis of brucellosis can be effective in the achievement of satisfying treatment results and prevention of chronic infections. The present study aimed to compare the efficiency of conventional microbiological and serological approaches with nested Polymerase chain reaction (nested PCR) for rapid diagnosis of human brucellosis. A total of 120 subjects with symptoms of brucellosis were included in the study. The sensitivity and specificity of nested PCR for the detection of Brucella bacteria were compared with serological and blood culture methods. Out of 120 patients enrolled, brucellosis was detected in 73 (60.83%) cases based on serological tests with a blood culture confirmation in 8.33% of participants. Based on the obtained results, 55% of cases were positive in serum agglutination test (SAT≥1:160), and Coombs (C-SAT≥1:160) tests. Furthermore, seven negative SAT cases were positive in C-SAT as evidence of chronic brucellosis. The results of the 2-mercaptoethanol (2-ME) ≥ 1:80 test were negative in six SAT-positive cases. Based on nested PCR results, 68.18% and 56.06% SAT positive samples were also detected by blood nested PCR and serum nested PCR, respectively. The sensitivity of blood nested PCR was significantly more than serum nested PCR, SAT≥1:160, and blood culture (P<0.001). Moreover, the specificity of blood and serum nested PCR was obtained at 100%, compared to blood culture and SAT≥ 1:160. In the present study, the nested PCR was able to identify chronic brucellosis in SAT negative patients. As evidenced by the obtained results, the nested PCR showed higher efficiency for rapid diagnosis of human brucellosis, as compared to the blood culture method. Furthermore, the findings pointed to the high performance of nested PCR for rapid diagnosis of both chronic and acute brucellosis.

Protozoan parasites of the genus Theileria are tick-borne parasites that have been found in many species of mammals. More than a dozen species of Theileria have been found in cattle, water buffalo, sheep, and goats. Theileria orientalis is a non-pathogenic blood protozoan parasite that was detected and identified during a regular investigation of piroplasmida infection in indigenous cattle in the spring of 2019 in Northern Provinces of Iran. In total, 92 blood samples were collected from different areas of Guilan and Mazandaran Provinces, Iran during the spring. The Giemsa stained blood smears did not show any parasitic infection; however, T. orientalis was identified by 18S rRNA gene polymerase chain reaction (PCR) and DNA sequencing. The specific sequenced DNA for T. orientalis was registered in GenBank under the accession number MN453385. The partial 18S rRNA gene sequence of the obtained DNA showed 100% nucleotide identity with reference sequences for the T. orientalis that have been registered from Europe, Africa, and Asia. Additionally, molecular phylogenetic studies have shown that T. orientalis Iran GC98-01 isolate belongs to nonpathogenic T. orientalis type 3 (buffeli). In this study, the indigenous Bos indicus cattle were detected as asymptomatic carriers of Theileria spp. infection. Here, we identified and genotyped T. orientalis for the first time as T. orientalis type 3 (buffeli) in Iran using molecular phylogenetic analysis and registered the 18S rRNA gene sequence of the T. orientalis GC98-01 isolate in GenBank.  Moreover, rare T. annulata infection was detected in cattle using semi-nested PCR in Mazandaran (Miankaleh peninsula). The T. orientalis can be differentiated from other Theileria and Babesia haemoprotozoan parasites by specific molecular assays.

Fascioliasis is an emerging and important food and water-borne disease in human communities which has become one of the most important health challenges in countries, like Iran. It causes weight loss, a decrease in feed conversion ratio as well as milk and meat production, and also reduces fertility in animals the prevalence of fasciolosis is increasing in some regions of the world due to various factors. Different methods have been used for the detection of Fasciola hepatica in animals. This study is the first to detect F. hepatica in Lori sheep using polymerase chain reaction (PCR) and conventional diagnostic methods in Western Iran. During three months, 195 fecal samples were collected from sheep in Lorestan province, Iran, using the stratified random sampling method. The conventional diagnostic methods, including wet mount microscopic examination and concentration assays, as well as the PCR technique targeting the intergenic spacer gene of F. hepatica, were used for the detection of the parasite in sheep. In total, 4 (2.1%) out of 195 examined stool samples were positive for F. hepatica based on the conventional assays. The PCR test was positive for F. hepatica in7 (3.6%) samples of 195 studied specimens. Statistical analyses of the data revealed that there is a significant difference between the results of diagnostic methods for F. hepatica detection (P=0.0421). Finally, the results showed that PCR has more diagnostic sensitivity, compared to conventional diagnostic methods, including the concentration techniques and microscopic examination. Hence, it can be advised to use PCR for the detection of F. hepatica in sheep.

This study aimed to investigate the fungal species isolation and confirmation by the Multiplex PCR method in aquatic fish. Evaluation of the inhibitory effect of nano-essential oils of Carum copticum on isolated fungal species was also conducted in this study. The PCR results showed that 3 out of 5 samples were diagnosed with Fusarium solani, and two of them were positive for Saprolegnia. Moreover, in 0.1% of the females' nanoparticles, one peak appeared that showed a particle with an average diameter of 360 nm, and two nanoparticles showed a peak with a mean diameter of 242 nm. The results of minimum inhibitory concentrations (MIC) and minimum fungicidal concentrations (MFC) showed that 0.01% nano essential oil had 0.08 and 0.07 mg/ml MIC values against Fusarium solani and Saprolegnia, respectively. Gram/ml was on the growth of Fusarium solani species. The essential oils of female plants had an MIC of 0.07 in 0.1% essential oil and 0.03 mg/ml in 0.01% essential oil in Saprolegnia. Furthermore, in the case of 0.1% nano essential oil, the results showed the MIC values of 0.04 and 0.03 mg/ml against Fusarium solani and Saprolegnia, respectively. The MFC values of 0.1% nano essential oil were 0.1 and 0.07 mg/ml against Fusarium solani and Saprolegnia,respectively. It was not found on Fusarium and Saprolegnia. Overall, the results of this study using PCR for direct detection showed that 70% and 50% of the samples were Fusarium solani and Saprolegnia positive, respectively; therefore, the PCR was an efficient method for the detection of fungi. According to the results of nano-essential oil (0.1%) of females, this nano-essence had a strong inhibitory effect on Fusarium solani and Saprolegnia.

The present study investigated the phylogenetic relationship based on cytochrome b gene sequences among pathogenic Theileria species (spp.) in Iran, including Theileria annulata and Theileria lestoquardi, along with other data available in GenBank. A total of 136 (cattle) and 80 (sheep) blood samples suspected of piroplasm infection were obtained from six different provinces of Iran. Both microscopic and molecular methods using species-specific primers were used for screening T. annulata and T. lestoquardi positive samples. Finally, the partial cytochrome b gene of 30 T. annulata and 5 T .lestoquardi were amplified, sequenced, and deposited in GenBank. The results indicated that there were 12 different genotypes among T. annulata isolates, while only one genotype was observed among T. lestoquardi isolates. T. lestoquardi infection in cattle was detected in one sample, and no T. annulata and T. lestoquardi coinfection were detected in sheep and cattle. In the phylogenetic tree, different Theileria spp. were placed in separate clades, and the reliability of depicted tree and monophyly of T. annulata and T. lestoquardi ingroups were supported by the bootstrap value of 94% which significantly indicated that these two species evolved from a common ancestor. The tree also showed that these two pathogenic spp. shared a more recent common ancestor, compared to another species of Theileria parasites. To the best of our knowledge, this study is the first phylogenic analysis of pathogenic Theileria spp. in Iran based on the cytochrome b gene sequences. In addition, the first T. lestoquardi cytochrome b gene was sequenced and deposited in GenBank.

The keratinolytic activities of dermatophyte species are accompanied by the secretion of enzymes, such as serine proteases, which are coded by the Subtilisin (SUB) genes. This study aimed to determine the presence of the SUB genes in the clinical and nonclinical samples of Trichophyton verrucosum and Microsporum gypseum. Isolation was carried out by direct and laboratory examination. Following that, for the determination of the presence of the SUB gene, polymerase chain reaction with specific primers was conducted. The frequencies of the SUB gene were observed in almost 66% of the isolates. Statistical analysis showed a significant relationship between the presence of the SUB gene and the samples collected from human, animals, and soil (p ˂0.005). The current investigation has been the first study of the presence/absence of the SUB gene in the clinical and nonclinical isolates of T. verrucosum and M. gypseum in Iran which may be a new step to perform further studies.

About half of the world's population is infected by Helicobacter pylori, which is related to various diseases. The increase in the resistance of H. pylori to antibiotics is alarming and requires new medication candidates. In this study, 83 acidic soil samples (pH 3.9-6.8) were collected from tea and rice farms, located in the semitropical strip in the north of Iran (Lahijan and Fooman cities, Gilan Province). After various pretreatments, including dry heating (120 oC, 10 min), exposure to electromagnetic waves (800 Hz, 3 min), and centrifuging (2950 g, 15 min), 33 acidophilic or acid-tolerant actinobacteria were isolated and their potentials as a source of active metabolites against H. pylori were investigated. According to phenotypic and molecular identification tests, the actinobacterial isolates were classified into Streptomyces and Kitasatospora genera. Among 10 strains that had anti-H. pylori activity, the highest potentials were seen in the strains UTMC 3061 and UTMC 3318. The minimum inhibitory concentrations (MIC) of the related metabolites were 125 and 62.5 µg/ml, respectively. In the checkerboard test, the metabolites of these actinobacteria showed synergism with clarithromycin and reduced its MIC from 1 to 0.5 µg/ml. However, no synergism was seen between the metabolites and amoxicillin or metronidazole. The gas chromatography-mass spectrometry (GC-MS) analysis of the metabolites showed some antimicrobial agents, including carbamic acid, maltol, 2.4-di-tert-butylphenol, methyl dimendone, prolylleucyl, and oleamide. The strains UTMC 3061 and UTMC 3318 showed 99.41 and 100% similarity in 16S rRNA gene sequence to Streptomyces spinoverrucosus and Streptomyces cirratus, respectively. Their metabolites showed good antibiotic activity and limited toxicity and can be considered as promising sources of natural products against H. pylori.

In the last couple of years, a number of new and rapid tests for the diagnosis of Tuberculosis (TB) have been developed based on the low molecular weight antigens from Mycobacterium tuberculosis (Mtb) culture supernatant. This study aimed to isolate and purify low molecular weight antigens secreted by Mtb strain C for diagnostic purpose. The secretory proteins from culture filtrate of Mtb were extracted using ammonium sulphate precipitations and sephadex-G50 gel chromatography. The obtained antigen fractions were analyzed for their protein concentrations and approximate molecular weight using Lowry method and SDS-PAGE (12.5%), respectively. DOT-ELISA and Western blot assay was performed to confirm the presence of purified low molecular weight proteins isolated from Mtb using sera from pulmonary tuberculosis patients (polyclonal antibodies). During chromatography, low molecular weight proteins were separated, that was approximately 0.7 mg/ml of the total proteins (1.662 mg/ml). The purified protein fractions in molecular weight range of 14 kDa-41kDa appeared during SDS-PAGE analysis. The chromatographic band fraction in the weight range of 30-41 kDa was identified in the TB patients’ sera using Western blotting. The low molecular weight proteins in the culture filtrate of Mtb strain C were purified using ammonium sulphate and chromatography. These fractions were confirmed using Western blotting. The obtained results might support the hypothesis that the Mtb culture filtrate antigens could be used as a rapid and sensitive assay for the detection of patients with pulmonary TB.

It is necessary to understand the frequency of virulence factor-encoding genes in the assessment of the carriage proportion. Moreover, it is required in the characterization of major unique antigens that are useful in the development of effective immunological-based preventive measures. The current study aimed to evaluate the frequency of three encoding-virulence genes associated with Enterotoxigenic (ET) and Shigatoxigenic Escherichia coli (E. coli/EC) pathotypes (k99, stx1, and stx2) in North of Sistan and Baluchistan Province, Iran. The frequency of k99, stx1, and stx2 was determined via polymerase chain reaction among E. coli isolates collected from the feces of the clinically healthy suckling (n=50) and diarrheic calves (n=50). The k99 gene was absent in all isolates, and the frequencies of the E. coli containing stx1 and stx2 or both stx1 and stx2 were estimated at 8%, 14%, and 4%, respectively, in the clinically healthy suckling calves (P>0.05), compared to 24%, 16%, and 6% in diarrheic animals (P<0.05). Among the three studied genes, there was a statistically significant difference between clinically healthy suckling and diarrheic calves in terms of the frequency of E. coli isolatescontaining stx1. On the other hand, the results of this study indicated that k99 was not a major fimbrial antigen-encoding gene in the ETECpopulation in the region. It is assumed that in any health measure intended to control the pathogen, other genes involved with encoding fimbriae should also be considered. The noticeable high frequency of E. coli isolates bearing stx1 and/or stx2 virulence elementsboth in clinically healthy and diarrheic suckling calves in this study isa concern for public health. Accordingly, it is recommended that further epidemiological studies be conducted on the role of the stx1 gene in the diarrhea of suckling calves in Sistan and Baluchistan Province, Iran.

Hyalomma spp. is responsible for the transmission of protozoan, bacterial, rickettsial, and viral diseases and causes huge economic loss to the livestock industry.Recently, there is a wide number of promising attempts to evaluate and use herbal preparations for ticks control.This study aimed to evaluate the acaricidal activity of aqueous and ethanol extracts of Colchicum autumnale (C. autumnale) rhizome and leaf against the Hyalomma spp. in vitro. The acaricidal activities of the Colchicum leaf aqueous (CLA), Colchicum leaf ethanolic (CLE), Colchicum rhizome aqueous (CRA), and Colchicum rhizome ethanolic (CRE) extracts were evaluated at concentrations of 50, 100, and 150 mg/ml and controls (distilled water and Cypermethrin) following 0.25, 0.5, 0.75, and 1 h of exposure. It is worth mentioning that the spraying method was used in these experiments. Data were analyzed through GraphPad Prism 5 software. In addition, the chemical composition of aqueous leaf extract was analyzed using gas chromatography-mass spectrometry (GC–MS). The carbamodithioic acid (30.04%) was the major chemical constituent identified. Based on the results, CLA, CLE, CRA, and CRE extracts had an acaricidal effect; however, this effect was more potent in CLE. The CLE extract showed a 100% mortality rate at 50, 100, and 150 mg/ml concentrations and 1 h of exposure. The effectiveness of CRA on the Hyalomma spp. was very low. The median lethal concentration (LC50) values were obtained at 100 mg/ml. The results indicated that C. autumnaleleaves contained potent acaricidal ingredients and might provide new acaricidal compounds for the effective control of Hyalomma spp. However, further studies are required to evaluate the efficacy of C. autumnale in vivo.

Fish represents one of the major sources of animal proteins, and different species of fish are susceptible to infections with parasites which cause severe tissue damage and cell destruction of the infected organ. Therefore, in 2019, this parasitological study was conducted to assess the helminth parasites infecting the African sharptooth catfish Clarias gariepinus that were collected from Lake Manzala, Egypt. Only nematode parasite was reported as a prevalent infection from the fish stomach with an infection rate of 7.5%. Depending on the seasonal prevalence, the extent of the infection was analyzed. It was indicated that parasite infection was only reported as 15% in the winter season. Morphological and morphometric analyses of the present parasite species revealed that it possesses all the characteristics of the Camallanus genus, whereas it is closely related to Camallanus polypteri described previously. It is characterized by the presence of a buccal capsule with longitudinal internal ridges, some of which are very short and ranged from 8-14 in males and 8-9 in females. The esophagus consisted of muscular and glandular portions, the middle position of the excretory pore to the muscular esophagus, the anterior location of deirids to the nerve ring, posterior end of males with two unequal spicules and caudal papillae; nonetheless, it is smooth and straight in females. In addition, some morphology and measurement differences for the different body parts were identified with other Camallanus species. Therefore, the present study can provide a full morphologically re-description of Camallanus polypteri with a new geographical location in the Egyptian freshwater.

The present study investigated the fine structure of amphids and phasmids, cuticle, muscles, and digestive tracts of Toxocara canis using optical and electron microscopy, hematoxylin-eosin (H&E) staining, and other specific stains. A number of 38 adult T.canis worms were obtained from the animal shelter of Urmia, and their small intestines were fixated in acidified formal alcohol and 10% formalin solutions. The anterior and posterior parts of male and female T.canis worms were prepared and cut at a thickness of 4-5 μm according to the conventional method in the histological laboratory. The samples were then stained using H&E and specific periodic acid-Schiff, Masson's trichrome, and Orcein staining. The structure of amphid (anterior), phasmid (posterior), cuticle, muscles, and digestive tracts of male and female worms were studied under light microscopy. Basal, intermediate, cortex, and cuticle surface coating of the parasite were visible. Alae were also observed as the thickenings in the cuticle. The muscle layer structure consists of non-branched cylindrical cells. The intestinal tract is composed of cuticular cogs, the esophagus is of filamentous-muscular structure, and the intestine is made of columnar epithelial tissue with microvilli and glycocalyx. The amphid structure consisted of cuticular protrusions with penetrations of the cephalic framework into their inner layers. Phasmid structure also includes protrusions in the cuticle and invagination of sensory neurons. It was concluded that for the most part, the histological structure of the cuticle can be studied by optical microscopy. The muscle structure in this parasite was very similar to the skeletal muscle in mammals. Furthermore, the epithelial structure of the intestine in this parasite was largely similar to the intestinal epithelium in mammals. Finally, regarding the amphid and phasmid structures, it was observed that they were protrusions covered by cuticles where neural, filamentous, and muscular structures were the core of these protrusions.

The present study aimed to assess the effect of tissue hypoxia induced by sodium cyanide (NaCN) on male mice fertility and the protective role of ethyl pyruvate (EP). A number of 30 adult mice were assigned to three groups: 1) a control group, 2) a treatment group treated with 2 mg/kg of NaCN, and 3) a treatment group treated with 2 mg/kg of NaCN, along with 40 mg/kg EP (NaCN+EP). After 35 days, animals were anesthetized and serum, sperm, and tissue samples were taken. The results demonstrated a significant decrease in sperm quality, reproduction potency, and anti-oxidant potential, as well as an increase in lipid peroxidation in the NaCN group (p <0.05). Moreover, the use of EP effectively restrained the disastrous effects of tissue hypoxia. It can be concluded that EP can moderate the complications resulting from tissue-hypoxia that is related to testes parameters.

This study conducted two experiments to evaluate the effects of two cheap adsorbents, including bentofeed (a commercial name of bentonite) and Persian melon peel biochar (PMPB) on the decolorization of water contaminated by methylene blue (MB) and ruminal fermentation pattern. The decolorization efficiency of bentofeed and PMPB at three levels of 0, 4, and 8 mg per 10 ml of 0, 3, 6, and 9 mg/L MB solutions mg/L after 3 and 24 h of incubation was evaluated by its absorbance at 660 nm. At all dye concentrations, PMPB, and bentofeed showed high potential in removing MB from water with an efficiency of 60%-99.5%. In both incubation times, the addition of 8 mg bentofeed had the highest effect on the removal efficiency when the dye concentration was 6 or 9 mg/L. However, the removal efficiency was declined with increasing MB concentration (p <0.05). Experiment two evaluated the effects of various levels (same as experiment one) of MB, bentofeed, and PMPB on in vitro gas production (GP) and volatile fatty acids (VFA) in two individual 4×3 factorial experiments. The potential GP (b), rate constant of gas production (c), metabolizable energy, organic matter digestibility, and total VFA were significantly decreased with increasing MB in the medium (p <0.05), while all parameters were increased when bentofeed or PMPB was added to the medium containing MB (p <0.05). The amounts of acetic, propionic, and butyric acids were not affected by PMPB; however, they changed when bentofeed was added to the medium (p <0.05). The NH3-N concentration was decreased significantly following the increase of MB; moreover, it was increased when PMPB and bentofeed were added to the medium. MB, as a water contaminant agent, had negative effects on ruminal fermentation parameters. Both adsorbents (i.e., PMPB and specially bentofeed) were efficiently able to remove MB from the water. The negative effects of MB on fermentation parameters were also alleviated as a result of using bentofeed or PMPB. It seems that bentofeed has the higher adsorption property of MB, compared to that of the PMPB.

Central dopaminergic (DAergic) and adrenergic systems have a prominent role in appetite regulation; however, their interaction(s) have not been studied in neonatal layer chickens.Therefore, the current study aimed to determine the interaction of central DAergic and noradrenergic systems in food intake regulation in neonatal layer chickens. In the first experiment, chickens received the intracerebroventricular (ICV) injection of a control solution, prazosin (i.e., α1 adrenergic receptor antagonist; 10 nmol), dopamine (DA; 40 nmol), and prazosin plus DA. The second to fifth experiments were similar to the first experiment except that the birds were injected with yohimbine (i.e., α2 receptor antagonist; 13 nmol), metoprolol (i.e., β1 adrenergic receptor antagonist; 24 nmol), ICI 118,551 (i.e., β2 adrenergic receptor antagonist; 5 nmol), and SR59230R (i.e., β3 adrenergic receptor antagonist; 20 nmol) instead of prazosin. In the sixth experiment, the chickens received ICV injection with the control solution and noradrenaline (NA; 75, 150, and 300 nmol). In the seventh experiment, the birds were injected with the control solution, SCH23390 (i.e., D1 DAergic receptor antagonist; 5 nmol), NA (300 nmol), and SCH23390 plus NA In the eighth experiment, the control solution, AMI-193 (i.e., D2 DAergic receptor antagonist; 5 nmol), NA (300 nmol), and AMI-193 plus NA were injected. Then, cumulative food intake was recorded at 30, 60, and 120 min after the injection. According to the obtained results, the ICV injection of DA (40 nmol) significantly decreased food intake in comparison to that reported for the control group (p <0.05). The co-injection of yohimbine plus DA significantly amplified DA-induced hypophagia in the neonatal chickens (p <0.05). In addition, the co-administration of ICI 118,551 plus DA significantly inhibited the hypophagic effect of DA in the neonatal chickens (p <0.05). Furthermore, NA (75, 150, and 300 nmol) significantly reduced food intake in a dose-dependent manner (p <0.05). The co-injection of SCH23390 plus NA decreased the hypophagic effect of NA in the neonatal chickens, compared to that reported for the control group (p <0.05). The co-injection of AMI-193 plus NA diminished NA-induced hypophagia, compared to that reported for the control group (p <0.05). The aforementioned results suggested that there is an interconnection between central DAergic and noradrenergic systems through α2/β2 adrenergic and D1/D2 DAergic receptors in food intake regulation in neonatal chicks.

Genistein (GEN), a soybean isoflavone, is structurally and functionally similar to endogenous estrogen; therefore, it has the potential to enhance estradiol properties. This study aimed to evaluate the effects of GEN on the reproductive performance and bone status of laying hens. In total, 80 Hy-line W-36 (40 weeks old, the late stage of egg production cycle) with an initial body weight of 1,230±15.8 g (Mean±S.E.M), similar egg production, and egg weight were randomly assigned into two groups with 10 replicates and 4 birds in each replicate (40 laying hens per group). Laying hen diets had 0 (control) and 20 mg/kg GEN (white powder, Sichuan Guanghan co. Ltd., purity of 98.5%) for 6 weeks (41 to 46). At the end of the experiment, 20 hens (one hen from each replicate) were slaughtered, and the samples of bone and shell gland (approximately 50 mg) were surgically taken immediately after slaughter for Real-time PCR. The results indicated that dietary GEN increased egg production, feed intake, and egg mass; however, it decreased egg weight (p <0.05). Furthermore, the feed conversion ratio was greater in birds received GEN, compared to those in the control group (p <0.05). GEN enhanced egg quality indices included eggshell strength, thickness, and percentage (p <0.05). Mechanical properties of the tibia, such as weight, length, and breaking strength were also increased by GEN (p <0.05). Moreover, dietary GEN increased the calcium content of the tibia (p <0.05). The mRNA expression of Calbindin-D28k (CaBP-D28k) and transient receptor potential vanilloid channel type 6 (TRPV6) upregulated in eggshell glands of hens treated with GEN paralleled to the controls (p <0.05). In conclusion, the findings of the present study showed that GEN had the potential to improve the bone physical characteristics, mineralization, and the productive performance of Hy-line W-36 laying hens in their post-peak period.

The current study aimed to determine the effect of metformin (MET) on histopathologic evaluation and antioxidant enzyme activity in experimental varicocele-induced rats. A total of 60 rats were randomly divided into six experimental groups. Group 1 (control) received no medication and underwent no surgery. In group 2 (sham), the rats received no medication and the abdominal cavity was opened; however, there was no varicocele induction. In group 3 (varicocele), the abdominal cavity was opened and the rats underwent varicocele induction and received no medication. In group 4, the abdominal cavity was opened and the animals received 25 mg/kg of MET for 42 days and were varicocele-induced. Groups 5 and 6 were similar to group 4 except that the animals received 50 and 100 mg/kg of MET, respectively. At the end of the 21st and 42nd days, the rats were euthanized and the left testis was removed for histological analysis and measurement of superoxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GPx), and total antioxidant status levels. According to the results, a dose-dependent difference was observed in testis damage grade in the MET treated groups, compared to that reported for the varicocele group (p <0.05). No difference was observed between 25 and 50 mg/kg of MET (P>0.05). Tissue MDA levels significantly increased in varicocele rats (p <0.05); however, MET (25, 50, and 100 mg/kg) in a dose-dependent manner decreased varicocele-induced MDA (p <0.05). Experimental varicocele significantly decreased SOD activity, compared to that reported for the control group (p <0.05). The administration of MET (25, 50, and 100 mg/kg) significantly increased tissue SOD activity in varicocele rats (p <0.05). The MET (25, 50, and 100 mg/kg) in a dose-dependent manner increased GPx activity in varicocele rats (p <0.05). There was no difference in MDA, SOD, and GPx levels between 25 and 50 mg/kg MET groups (P>0.05). The aforementioned findings suggested that MET treatment had beneficial effects on varicocele.

Spiders are one of the most important orders of Arachnida comprising more than 48,000 species in the world. Except for families Uloboridae and Holarchaeidae, all others are classified as the venomous spider. However, only about 200 species are medically relevant and cause public health problems or even death. In Iran, there are 51 families and 763 species of spiders, of which the families, Theridiidae and Sicariidae are dangerous for the human being, and the first one is more prevalent. The Latrodectus is considered one of the most poisonous spiders in the world and Iran. This genus has five species in Iran, among which Latrodectus tredecimguttatus (black widow spider or “Dolmak”) is considered one of the most poisonous spiders in Iran. Therefore, this study aimed to investigate this species in the Northwest of Iran (West Azerbaijan, East Azerbaijan, and Ardabil provinces, Iran). Spatial distribution maps were prepared using GIS 9.4. In the current study, five adult female spiders were collected from Germi and Ardabil cities (Ardabil Province), Ahar County (East Azerbaijan province), and Urmia city (West Azerbaijan province) of Iran.  These species were first observed in Ardabil province, Iran. Therefore, the presence of Latrodectus species under the rocks in wheat farms in this corner of Iran may be a threat to farms and visitors. People in these areas should wear gloves and avoid any activity that disturbs the spiders and make them aggressive.

Leptospirosis is a zoonotic disease with global importance, and the animals are the source of transmission of this disease through shedding in their urine. Accordingly, it is essential to conduct epidemiological studies of leptospirosis in order to diagnose this disease in dogs and reduce the risk of transmission to humans. This study aimed to perform a seroepidemiological analysis of Leptospiral infection in stray dogs in Alborz, Iran, using the Microscopic Agglutination Test (MAT). In total, 110 blood samples were collected from stray dogs to detect the antibodies against leptospira interrogans serovarsby the MAT.  The prevalence rate of positive MAT tests in stray dogs was estimated at 21.84%. The following protocol confirmed that the most common titers were 1:200 (50%) and 1:400 (25%). In addition, the most prevalent Leptospira serovars were L. Canicola (33.33%), and the lowest belonged to L. Pomona (4.1%). Moreover, no significant difference was observed between the age and gender of the dogs regarding their MAT titer (P>0.05). The results also showed a high prevalence of leptospirosis in stray dogs of Koohsar in Alborz province, Iran. Since Leptospirosis is a zoonosis disease, it should be studied continuously in humans and animals, especially dogs.

Cystic fibrosis (CF) is a genetic disease with a high rate of morbidity and mortality. Children with CF commonly suffer from recurrent and persistent pulmonary tract infections caused by diverse bacterial pathogens. This study aimed to investigate the prevalence, antimicrobial susceptibility, and biofilm formation of bacterial isolates in pediatric patients with CF. The study population of this cross-sectional study included 8,908 children suspected to have CF by clinical manifestations from March 2015 to August 2017 who were referred to the Tehran Pediatric Central Hospital, Iran. The tests carried out for each participant included screening sweat test, sputum culture, antibiotic susceptibility test using Kirby-Bauer disk diffusion method, and biofilm formation in microtiter plates method. Based onclinical examination and screening sweat test, 183 (2.05 %( out of 8,908 children, were positive for CF. The mean age of children was estimated at 2.93 years, and the majority of them were male (n=103, 56.2%). No gender-specific difference was observed in CF disease in this study (P>0.05). In addition, the results of sputum culture showed that 153 (83.6%) microorganisms (bacteria and fungi) were collected from CF patients. Normal flora was isolated in 30 (16.4%) patients and more than one bacterial species were isolated in 7.2% of patients. The obtained results indicated that Pseudomonas aeruginosa was the most prevalent isolated bacteria followed by Staphylococcus aureus, and Klebsiella pneumoniae. Based on the antibiotic susceptibility test results, P. aeruginosa and piperacillin/tazobactam had the highest (11.7%) and the lowest (2.3%) resistance rate against gentamicin, respectively. However, all K. pneumoniae isolates were resistant to Cefotaxime. Among S. aureus isolates, 83.4% and 16.6% were methicillin-susceptible Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus respectively. Concerning biofilm formation, 76%, 67%, and 72.5% of P. aeruginosa, S. aureus, and K. pneumoniae isolates were biofilm producers, respectively. Based on the study results, P. aeruginosa was the dominant pathogen in pediatric patients with CF from Tehran, Iran, and most of the pathogens were biofilm producers. No severe antibiotic resistance was observed in the isolates; however, the anti-microbial resistance profile should be carefully checked in CF patients on a regular basis.

Malignant melanoma is a neoplasm that originates from melanocytes. This tumor is observed in cutaneous and non-cutaneous forms, and it is considered one of the most life-threatening types of cancers. Non-cutaneous melanoma is a complex of unique and malignant complications that are easily separable from cutaneous type. Since the ultraviolet radiation from the sun damages DNA and is an oxidative stress factor in melanoma and there are more melanocytes in the basal layer of skin than other parts of the body, the cutaneous form has more prevalence. Most of the time, non-cutaneous form is the result of cutaneous metastasis but both forms can occur primarily. Furthermore, non-cutaneous form usually happens in mucosal layers, intestines, and eyes; moreover, the main reasons are ectopic melanocytes or their unwanted regressive growing. Malignant melanoma can occur in all domestic animals; however, they seem to be rare in sheep and goats. Herein, we describe a rare case of the primary non-cutaneous form of malignant melanoma in a three-year-old indigenous female goat. During meat inspection procedures in a slaughterhouse in Tabriz, Iran, we encountered numerous round firm black masses on visceral surfaces and serous membranes of the abdominal and thoracic cavities. The liver and lungs were prominently affected. Samples were taken from involved parts, and malignant melanoma was confirmed in the histopathological examination due to pleomorphism and polymorphism and melanin pigments in cells with eosinophilic cytoplasm. According to what was stated in the "manual on meat inspection for developing countries", the carcass was not convenient for human use and condemned by the inspector.