Composition Comprising Parasite Eggs And Methods For Isolation And Storage Of Parasite Eggs

Abstract

The invention relates to a composition for storage and development of eggs from helminthic parasites, where the composition further comprises a liquid carrier having a pH value of below 7 at a temperature of from 10 C to ambient temperature. The liquid carrier can be sulphuric acid, H 2 SO 4 with a pH in the range of from 0 to 2, and antibiotics can be added. The invention further relates to a method for treating, ameliorating, prophylactic or curative, an autoimmune or allergic disease in an individual, animal or man, using eggs being separated from the composition. The present invention also relates to methods for isolation, embryonation and preservation of eggs of helminthic parasite, and to a method for producing a pharmaceutical composition comprising a helminthic parasite preparation. The helminthic parasite eggs may be from the pig whipworm; Trichuris suis ova (TSO).

1 . A method for the isolation and storage of eggs from the pig whipworm  Trichuris suis,  said method comprising the steps of:
 a) isolating parasite eggs either using a method 1) comprising the steps of isolating eggs from worms which have been removed from the intestine of pigs and grown in a growth medium suitable for such growth, wherein the worms lay their eggs in said suitable growth medium, or a method 2) comprising the steps of using an infected animal, or infecting an animal, such as a pig, with  Trichuris suis  and isolating eggs from fecal material produced by said pig, and   b) storage of the isolated and optionally cleaned eggs in an acidic medium, such as H 2 SO 4 , such as H 2 SO 4  having a pH of 0-2, eventually with antibiotics, to allow for the development of the eggs (embryonation) and to inactivate any contaminating bacteria, fungi and viruses.   
   
   
       2 . A method for the isolation and storage of eggs from the pig whipworm  Trichuris suis,  said method comprising the steps of:
 a) isolating parasite eggs either using a method 1) comprising the steps of isolating eggs from worms which have been removed from the intestine of pigs and grown in a growth medium suitable for such growth, wherein the worms lay their eggs in said suitable growth medium, or a method 2) comprising the steps of using an infected animal, or infecting an animal, such as a pig, with  Trichuris suis  and isolating eggs from fecal material produced by said pig, and   b) filtering the isolated material to reduce particle contamination and eggs of foreign pig parasites,   c) floating the isolated material to reduce particle contamination,   d) washing the isolated material in an acidic medium, such as sulphuric acid, H 2 SO 4 , such as H 2 SO 4  having a pH of 0-2, to reduce the counts of foreign pathogens (bacteria, fungi and viruses) both by dilution and inactivation,   e) storing the isolated and cleaned eggs in an acidic medium, such as H 2 SO 4 , such as H 2 SO 4  having a pH of 0-2, eventually supplemented with antibiotics, to allow for the development of the eggs (embryonation) and to further inactivate any bacteria, fungi and/or viruses, if present,   f) filtering and floating the embryonated eggs to isolate a fraction of eggs that have the highest degree of embryonation (biotic potential and a pharmaceutical raw material), and   g) storing the isolated and embryonated eggs in an acidic medium, such as H 2 SO 4 , such as H 2 SO 4  having a pH of 0-2, eventually supplemented by antibiotics, thereby allowing for maintenance of biotic potential and the prevention of pathogen growth.   
   
   
       3 . A method for producing a pharmaceutical composition comprising a helminthic parasite preparation, comprising the steps of:
 (1) raising a preparatory animal in a specific human pathogen-free environment;   (2) obtaining a helminthic parasite isolate from said preparatory animal;   (3) extracting eggs from said helminthic parasite isolate;   (4) storing un-embryonated eggs from said first helminthic parasite isolate in a composition further comprising an acidic liquid carrier;   (5) embryonating eggs from said helminthic parasite isolate under suitable conditions in said acidic liquid carrier to generate a pharmaceutical composition; and   (6) storing embryonated eggs from said helminthic parasite isolate under suitable conditions in said acidic liquid carrier.   
   
   
       4 . The method of  claim 3 , wherein the step of isolating a helminthic parasite comprises obtaining a stool from said preparatory animal, and isolating the helminthic parasite from said stool. 
   
   
       5 . The method of  claim 4 , wherein the step of isolating a helminthic parasite comprises removing a tissue from said preparatory animal, and isolating the helminthic parasite or its eggs from said tissue. 
   
   
       6 . The method of  claim 5 , wherein said tissue is an internal organ such as intestines. 
   
   
       7 . The method of  claim 5 , wherein the step of isolating said helminthic parasite further comprises the steps of:
 (1) dissecting the tissue from said preparatory animal to allow for macroscopical isolation of worms to produce a worm culture wherein the worms lay eggs,   (2) filtering the worm culture to produce a filtrate with eggs; and   (3) isolating the eggs from said filtrate, thereby extracting eggs from said helminthic parasite isolate.   
   
   
       8 . The method of  claim 3 , wherein the helminthic parasite preparation comprises a parasite which is a nematode. 
   
   
       9 . The method of  claim 3 , wherein the helminthic parasite preparation comprises a parasite selected from the group consisting of the genera  Ascaris, Enterobius, Trichuris, Ancylostoma, Necator, Strongyloides, Oesophagostomum, Toxascaris, Parascaris.    
   
   
       10 . The method of  claim 3 , wherein the helminthic parasite preparation comprises a parasite which is a platyhelminth. 
   
   
       11 . The method of  claim 3 , wherein the helminthic parasite preparation comprises a parasite selected from the group consisting of trematodes and cestodes. 
   
   
       12 . The method of  claim 3 , wherein the helminthic parasite preparation comprises a parasite selected from the group consisting of genera  Fasciolopsis, Echinostoma, Heterophyes, Clonorchis, Opistorchis, Fasciola, Schistosoma, Diphyllobothrium, Taenia  and  Hymenolepsis.    
   
   
       13 . The method of  claim 3 , wherein the helminthic parasite preparation comprises a parasite selected from the group consisting of filarial parasites and lung flukes. 
   
   
       14 . The method of  claim 3 , wherein the helminthic parasite preparation comprises a parasite selected from the group consisting of genera  Trichuris, Nippostrongylus, Heligmosomoides, Hymenolepsis, Angiostrongylus, Ascaris, Toxocara, Gnathostoma, Ancylostoma, Anisakis  and  Pseudoterranova, Oesophagostomum, Toxascaris,  and  Parascaris.    
   
   
       15 . The method of  claim 3 , wherein the helminthic parasite is  Trichuris suis.