Isolation of common carp ovarian follicular cells and evaluation of their endocrine activity in primary cell culture

To study viability and activity of isolated common carp, Cyprinus carpio, ovarian follicular cells (granulosa and theca cells), 17-α-Hydroxy progesterone (17α-OHP) and 17β-Estradiol (E2) levels were estimated in the culture media of cultivated carp ovarian follicular cells, using radioimmunoassay (RIA). Oocytes were isolated from the ovaries of female carp. Interstitial tissue was manually removed in order to obtain single oocytes surrounded only by the follicular envelope. Such a preparation was trypsinized at room temperature. Follicular cell suspension contained both cells and cell clumps. It was the mixture of theca (T) and granulosa (G) cells. The cells were grown as monolayer in 24-well microplates in M199 medium supplemented with FBS. Culture media were analyzed for estrogen and progesterone content by appropriate radioimmunoassay. Trypsinization of ovarian follicles resulted in the formation of the cell suspension which contained a mixture of G and T cells. The follicular cells attached to glass and grew well during culture period. E2 was the main steroid hormone secreted by cultivated cells. Estrogen secretion increased by 415.52 ± 25 pg/ml at the first 3 days up to 530.25 ± 55.8 pg/ml on day 5 and it didn't change significantly until the end of the experiment. 17α-OHP secretion, however, gradually increased from 23.84 ± 8.2 pg/ml at the beginning of culture up to 35.76 ± 5.4 pg/ml at the end of cultivating. As the result of the present study the fish follicular cells grown in tissue culture were steroidogenically active cells as expressed by the secretion of E2 and P4 and the E2 was a dominant hormone secreted by isolated follicular cells, which it correlated closely with vitellogenes stage

PCR-based molecular characterization of Toxocara spp. using feces of stray cats: a study from Southwest Iran

Abstract Feces of stray cat are potential sources of gastrointestinal parasites and play a crucial role in spreading and transmitting parasite eggs, larvae, and oocysts through contamination of soil, food, or water. In this study, we investigated the prevalence of Toxocara spp. infection in stray cats in Ahvaz city, southwest Iran. Eggs of Toxocara spp. in feces of stray cats were detected by the sucrose flotation method, and identification was conducted by polymerase chain reaction (PCR) and DNA sequencing. Of the 140 fecal samples that were randomly collected from public environments during the months of January to May 2012, 45% were found to harbour Toxocara spp. eggs. The highest prevalence of Toxocara spp. eggs was found in the central area of Ahvaz city (28.6%). T. canis eggs were found in 4 (6.34%) of the 63 positive samples. Stray cats are found in parks, playgrounds, and other public places and may be a potential contamination risk. Identification of Toxocara spp. using molecular methods is sufficiently sensitive to detect low levels of parasites and identify the different Toxocara spp. in feces. The relatively high prevalence of Toxocara spp. infection may continue to increase due to lack of effective environmental hygiene control in Iran. Consequently, there is a need to plan adequate programs to detect, identify, and control this infection as well as stray cats in the region

Stray Cats Gastrointestinal Parasites and its Association With Public Health in Ahvaz City, South Western of Iran

Background: Cats are the hosts for some zoonotic parasites such as Toxoplasma gondii and Toxocara spp. which are important in medicine and veterinary. Studies on the prevalence of intestinal parasites of cats have received little attention in south west of Iran. Objectives: The current study aimed to investigate the prevalence of parasites in stray cats in Ahvaz. Materials and Methods: Random sampling was carried out from January to May 2012. One hundred and forty fecal samples from stray cats were examined using sucrose flotation method. Results: Gastrointestinal parasites were found in 121 of the 140 (86.4%) examined samples. The parasites detected in stray cats were Toxocara spp. (45%, 63/140), Isospora spp. (21.4%, 30/140), nematode larvae (21.4%, 30/140), Taenia spp. (18.6%, 26/140), Sarcocystis spp. (17.1%, 24/140), Eimeria spp. (15%, 21/140), Blastocystis spp. (14.3%, 20/140), Giardia spp, (10.7%, 15/140), Physaloptera spp. (7.1%, 10/140), and amoeba cyst (5.7%, 8/140) respectively. The prevalence of infection by Joyexiella spp. and hook worms (4.3%, 6/140), for example, Dipylidium caninum (2.9%, 4/140) was similar; and the prevalence of infection by T. gondii and Dicrocoelium dendriticum was similar (1.4%, 2/140). Conclusions: Since the prevalence of zoonotic gastrointestinal parasites such as Toxocara spp. in stray cats is high, there is a need to plan adequate programs to control these zoonotic parasites

Introducing the best cell culture method for primary hepatocyte from orangespotted grouper, Epinephelus coioides

Liver is one of the most important organs in vertebrates that have important roles in detoxifying. This organ was used as a target organ in many physiological and toxicological aspects. The main purpose of the present study was developing appropriate methodology for the primary cultivation of hepatic cells from orange-spotted Grouper, Epinephelus coioides, a subtropical fish species of the family Serranidae. In present study, hepatocytes were isolated from five grouper individuals. Initially, the fish wiped with 70% ethanol. Liver were removed and cut into small pieces with scissors and hepatocytes were disconnected using different enzymatic digestion with collagenase (Type 1 and 4) and trypsin and additional nutrient materials in culturing mediums. Then, cells were cultured for 2 weeks in Lebowitz L-15 under 3 methods: 1. using enzymatic digestion by trypsin, 2. using enzymatic digestion by collagenase (type 1 and 4) and 3. Using nutrients and additives was cultured. Finally, effects of different incubation temperature (20, 25, 28, 30 and 32 degree of Celsius) and Bovine serum content (0, 10 and 20% and 20%+ITS) on cell growth were estimated. According to the results, digestion by collagenase type 4, resulted in more cell colonization and growth in comparison with other methods. At the same method, cells showed fibroblastic morphology. In conclusion, the best culture method for primary hepatocyte from orange-spotted Grouper, Epinephelus coioides, was using ITS+20%FBS under 30 degree of Celsius incubation temperature

The Study of Apoptotic Effect of p-Coumaric Acid on Breast Cancer Cells MCF-7

Introduction: Polyphenolic compounds have anti proliferative and induced apoptotic features on cancer cells. p-Coumaric acid can be abundantly found in fruits, vegetables, plant production and honey. .&nbsp; Breast cancer is the most frequently diagnosed cancer among women in the world. This study aimed to investigate the effect and mechanism of p- coumaric acid on apoptosis of MCF-7 breast cancer cells. Methods: In order to study appoptic effect of p- coumaric acid, MCF-7 breast cancer cells were treated with different concentrations of p- coumaric acid (10, 37, 70, 150 and 300 mM) for 24 h. Cell viability was determined using MTT assay. Apoptosis markers including phosphatidylserine exposure at the outer leaflet of the plasma membrane were measured using flow cytometery for Annexin V affinity. Results: Cell viability of MCF-7 cells was decreased with increasing of p- coumaric acid concentration. Maximal effect of p- coumaric acid was observed in cells that treated with 300 mM for 24h (p< 0.05). Viability assay showed that the IC50 of p- coumaric acid in MCF-7 cells was about 40 mM. p- coumaric acid at dose of 300 mM significantly increased the late apoptotic cells with Annexin V+ and propium iodide (PI+) features after 24 h treatment. Conclusion: The results of this study showed that p- coumaric acid had effective appoptic activity against MCF-7 cells. The results can be helpful in understanding the anticancer mechanism of p- coumaric acid and using it was suggested as an alternative or complementary drug in cancer chemotherapy

The allocation of crop production resources in the southeast of Iran: the application of the water-energy-food nexus approach

Introduction: Due to the expansion of population, economic progress, urbanization, increasing food demands, and diversification of food systems, resources are being excessively exploited and degraded. This is compounded by the challenges posed by climate change and limited resources, as well as inadequate management practices. The concept of water-energy-food (WEF) nexus management recognizes the interdependencies among various resources, such as water, food, and energy, in order to promote sustainable resource management. By establishing a harmonious balance among different objectives, this approach aims to safeguard the well-being of both human societies and the environment, ensuring the fulfillment of needs and the preservation of benefits for both parties.Methods: In this study, the water-energy-food (WEF) nexus approach is applied to the Sistan plain, located in the southeastern region of Iran, to effectively redistribute production resources within the agricultural sector. The methodology employed is multi-objective programming, which incorporates various goals. These objectives encompass maximizing farmer revenue and energy derived from food production (measured in calories), while simultaneously minimizing greenhouse gas (GHG) emissions, irrigation water consumption, and overall energy consumption throughout the 2018–2019 crop year.Results: The findings of this study demonstrate that implementing the water-energy-food (WEF) nexus approach in the Sistan plain yields positive outcomes. Despite a reduction in the cultivation area, there is a notable shift towards growing more nutritious crops. This shift not only contributes to food security but also increases crop calorie production from 457.16 million to 565.19 million. Consequently, there is a decrease in irrigation water consumption from 261.62 million to 260.48 million cubic meters, energy consumption from 1400.13 million to 1396.81 million MJ per hectare, and greenhouse gas (GHG) emissions from 0.014 million to 0.0139 million tons per hectare.Discussion: Analyzing the physical and economic productivity reveals that GHG emissions had the highest productivity in terms of both physical and economic measures in Zahak County. As the WEF nexus approach aims to preserve and prevent environmental degradation, it is recommended to implement development and bio-balance policies utilizing this approach to ensure environmental conservation

Comparison of Fecal Egg Counts and ELISA for the diagnosis of Dicrocoelium Dendriticum infection

From economical point of view, Dicrocoelium Dendriticum (D. dendriticum) causes a lot of damages to the livestock industry annually. So, the rapid diagnosis of infection is very important. The diagnosis is based on egg per count of feces (EPG) test because detection according to clinical symptoms is difficult. Since EPG is not accurate and sensitive, the serological methods become important for the diagnosis of this parasite as they are more accurate in comparison to EPG test and they are able to diagnose infection in a short time. In this study, somatic and Excretory-secretory antigens (EsAg) were isolated. The ELISA test was set up according to positive and negative sera and the results which were obtained compared to those obtained by the EPG test. The prevalence of infection in 550 samples by ELISA and EPG methods were 56% and 7% respectively, which shows the significant difference between these methods in examining the rate of infection. Based on the results, the specificity and sensitivity in ELISA test were 95% and 94%, respectively. The results showed that the ELISA is a more reliable test in comparison to EPG test for the rapid diagnosis of D. dendriticum infection