Characterization of Membrane-Associated Progesterone Receptor Component-2 (MAPRC2) from Trichinella spiralis and Its Interaction with Progesterone and Mifepristone

Trichinellosis is a foodborne zoonotic disease caused by Trichinella spp., including Trichinella spiralis. In the present study, T. spiralis membrane-associated progesterone receptor component-2 (Ts-MAPRC2) gene was cloned and characterized using protein sequencing analysis. Furthermore, the expression, purification, immunoblot assay, binding ability with progesterone antibody, and immunofluorescence assay were performed. A direct effect of progesterone (P4) and mifepristone (RU486) on the Ts-MAPRC2 gene was determined using in vitro cell culture that showed different expression levels at all developmental stages (muscle larvae (ML), female adult worm (F-AL), male adult worm (M-AL), and newborn larvae (NBL)). Subsequently, the in vitro phenotypic effects of P4, RU486, and rTs-MAPRC2-Ab on F-AL and ML stages were measured. Later, the in vivo phenotypic effect and relative mRNA expression of mifepristone on the F-AL stage were studied. Our results revealed that the Ts-MAPRC2 gene is critical to maintaining pregnancy in the female adult worm (F-AL) of T. spiralis. The 300 ng/mL of P4 and 100 ng/mL of RU486 showed downregulation of the Ts-MAPRC2 gene in F-AL (p ≤ 0.05). This plays an important role in abortion and possibly decreases the worm burden of T. spiralis in the host. Only 30 ng/mL P4 showed significant upregulation in F-AL (p ≤ 0.05). The current study provides new insights regarding the antihormone (P4 and RU486) drug design and vaccine therapy of recombinant (rTs-MAPRC2) protein as well as their combined effects to control T. spiralis infection.

Characterization of Membrane-associated Progesterone Receptor Component-2 (MAPRC2) From Trichinella spiralis and Its Interaction With Progesterone and Mifepristone

Background: Trichinellosis is a food-borne zoonotic disease caused by nematode viz., Trichinella spiralis. Physiologically, the high progesterone (P4) doses cause new borne larvae (NBL) mortality in the parasite, while the low doses maintain pregnancy. In contrast, Mifepristone (RU486) works as an antagonist against the progesterone receptor (PR) and possesses abortifacient activities. Methods: In the present study, T. spiralis membrane-associated progesterone receptor component-2 (Ts-MAPRC2) gene was cloned and characterized by protein sequencing. Furthermore, the expression, purification, immunoblot assay, binding ability with progesterone antibody, and immunofluorescence assay were performed. A direct effect of progesterone (P4) and mifepristone (RU486) on the Ts-MAPRC2 gene was determined using in-vitro cell culture that showed different expression levels at all developmental stages [muscle larvae (ML), female adult worm (F-AL), male adult worm (M-AL) and new borne larvae (NBL)]. Subsequently, the in-vitro phenotypic effect of P4, RU486, and rTs-MAPRC2-Ab on F-AL and ML stages were measured. Later on, the in-vivo phenotypic effect and relative mRNA expression of mifepristone on the F-AL stage were studied. Results: Our results revealed that the Ts-MAPRC2 gene is critical to maintaining pregnancy in the female adult worm (F-AL) of T. spiralis. The P300 ng/mL of P4 and M100 ng/mL of RU486 showed downregulation of the Ts-MAPRC2 gene in F-AL (P ≤ 0.05). This plays an important role in abortion and possibly decreases the worm burden of T. spiralis in the host. Only P30 ng/mL showed significant upregulation in F-AL (P ≤ 0.05). Conclusions: The current study provides new insights regarding the antihormone (P4 & RU486) drug design and vaccine therapy of recombinant (rTs-MAPRC2) protein as well as their combined effects to control T. spiralis infection.

L3 Populations in Laying Hens Infected with 6,000 L2 of Ascaridia galli

The aim of the present study was to determine the survival of L3 populations in intestine ofchickens exposed to experimental Ascaridia galli infection. Nature female adult worm were obtained fromlumen of village chickens in a comercial abattoir in Bogor. The eggs (L1) obtained from uteri female adultworms were incubated in sterile aquadestilata at room temperature for 10-20 days developed embrionatedeggs (L2). Five groups (A-D) of 80 head chickens were infected with, 6000 L2 A. galli respectively. Thechickens of group A were infected six times with dose of each 1,000 L2 with an interval of one hour. Thechickens of group B were infected three times with dose of each 2,000 L2 with an interval of two hours.The chickens of group C were infected six times with dose of each 3,000 L2 with an interval of three hours. The chickens of group D were infected one time with single dose 6,000 L2. A. galli L3 were recovered from intestines of 80 heads chickens seven days after oesophagus inoculation with 6,000 L2.The result showed that total 702,000 L1 and 628,000 L2 collected from 124 A. galli female adult worms.The percentage of L1 developed L2 is 89.46% and L2 developed L3 is 11.27%. Significant survival of L3higher populations in intestine of chickens observed only in the group D. The results indicated thatchickens infected high dose of A. galli caused the decrease of host defence against ascaridiosis. Keywords: Ascaridia galli, embrionated eggs, larvae