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

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

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