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PubMed1986

Evaluation of enzyme-linked immunosorbent assay in serological diagnosis of human neurocysticercosis using paired samples of serum and cerebrospinal fluid.

The applicability of micro-ELISA was evaluatd in human neuro-cysticercosis using paired samples of serum and CSF. A total of 355 cases who were mostly neurologic patients was subjected. Cystic fluid of C. cellulosae was used as antigen in protein concentration of 2.5 micro-g/ml. Serum was diluted to 1:100 and CSF was undiluted in the assay for the specific IgG antibody level. The differential criterion of the positive reaction was the abs. of 0.18 in both samples. The results were summarized as follows: The overall sensitivity of the micro-ELISA in 71 confirmed neurocysticercosis was 90.1%; the sensitivity by serum was 77.5% and that by CSF was 83.1%. CSF was a more sensitive and valuable material. Most of the false negative cases of neuro-cysticercosis showed far lower level of abs. rather than marginal. The overall specificity of the micro-ELISA in 52 confirmed other neurologic diseases was 88.5%; the specificities by serum and by CSF were 94.2% respectively. Cases of other neurologic diseases did not show false positive reactions in both samples. When serum was assayed, taeniasis(2/18), sparganosis(2/20), paragonimiasis(1/56), clonorchiasis(1/15) and fascioliasis(1/1) cases showed cross reactions. When CSF was assayed, 2 of 10 neuro-sparganosis showed cross reactions while none of 9 neuro-paragonimiasis showed it. Out of 71 confirmed neuro-cysticercosis cases, 6 and 11 showed cross reactions by serum and CSF to crude extract antigen of sparganum; but no case did show it to crude extract antigen of Paragonimus westermani. Ventricular CSF showed low or negative levels of IgG antibody than lumbar CSF unless the lesion was at the lateral ventricle itself. Out of 4 racemose cysticercosis cases, 3 showed positive reaction in serum while all of 3 examined CSF were positive. The above results indicated that the serological test for detecting the specific IgG antibody by micro-ELISA using paired samples of serum and CSF was very helpful for clinical differentiation of neuro-cysticercosis from neurologic diseases of other causes.

Journal Article↗
PubMed1993

Antigenic protein fractions of Metagonimus yokogawai reacting with patient sera.

Antibody test is sometimes necessary for the diagnosis of acute human metagonimiasis because eggs may not be detected in stool. The antibody test (ELISA) was evaluated for its significance by reacting human sera from clinically diagnosed metagonimiasis, fascioliasis, clonorchiasis and paragonimiasis with 4 crude extracts of Metagonimus yokogawai (metacercariae), adults of Fasciola hepatica, Clonorchis sinensis and Paragonimus westermani. By ELISA, 10 of 11 metagonimiasis sera showed the highest absorbance (abs.) to the homologous antigen. Cross reactions to M. yokogawai antigen occurred most frequently in clonorchiasis sera. The antigenic protein fractions in M. yokogawai metacercarial extract were observed by SDS-PAGE/immunoblot using patients and control sera together with experimental cat sera. Out of 14 protein bands in the extract, 11 bands were reacting. Cross reacting bands to other trematodiasis sera were frequently observed. Of the reacting bands, 66 and 22 kDa proteins were recognized as specific for metagonimiasis.

Animals↗
PubMed2006

Common conditions leading to cattle carcass and offal condemnations at 3 abattoirs in the Western Province of Zambia and their zoonotic implications to consumers.

From a total of 32 717 cattle slaughtered, 183 whole carcass condemnations were attributable to 9 diseases and conditions, namely, tuberculosis (TB), cysticercosis, emaciation, generalised lymphadenitis, jaundice, abscesses, moribund, sarcosporidiosis and odour. Bovine TB was the most important cause of condemnations (152/183, 83.1%). Bovine cysticercosis and sarcosporidiosis accounted for 5/183 (2.7%) and 8/183 (4.4%), respectively, while each of the remaining conditions contributed less. Among the many conditions responsible for offal/organ condemnations were fascioliasis, contagious bovine pleuropneumonia, hydatidosis and TB. In terms of number and weight, Fasciola gigantica infections made livers and lungs the most condemned offals (20.1% and 0.7%, respectively). Hydatidosis was the cause of 0.9% lung and 0.1% liver losses. Cysticercus bovis contributed to only 0.05% of all inspected tongues, hearts, and heads. TB was very rare in heads (0.01%). The financial impact of whole carcasses and offals condemned during the study period was enormous and deprived livestock farmers of the much needed revenue and consumers of protein sources. Much or all of the condemned material that could have been useful was wasted by not being retrieved for conversion to processed meat, bone meal or pet food. Failure to detect lesions of potential zoonotic diseases at slaughter poses a health risk to consumers especially when meat is eaten undercooked.

Abattoirs↗
PubMed1979

The enzyme-linked immunosorbent assay (ELISA) for the immunodiagnosis of schistosomiasis.

The enzyme-linked immunosorbent assay (ELISA) done with soluble egg antigens (SEA) of Schistosoma mansoni was utilized for the detection of infections with schistosomes. The method readily detected experimental infections in NIH outbred, New Zealand black, and New Zealand white mice by 6-10 weeks post-exposure to S. mansoni cercariae. In addition, the test was negative when the sera from 11 Puerto Rican normal controls were examined and was positive in 8 of 10 serum samples from humans with schistosomiasis mansoni. However, extensive cross-reactivity was seen when using serum from humans with fascioliasis, trichinosis, cysticercosis, and echinococcosis. Thus the ELISA test done with SEA as antigen lacks immunologic specificity. For the method to be an effective seroepidemiological tool in areas where these parasites are endemic further purification of the antigen and more extensive understanding of its components are needed.

Animals↗
PubMed1991

Evaluation of a monoclonal-antibody based antigen assay for diagnosis of Wuchereria bancrofti infection in Egypt.

Conventional methods for diagnosis of Wuchereria bancrofti infection are insensitive and often impractical because of the need for night blood collections. A sensitive and specific antigen detection assay has been developed for W. bancrofti, which is based on a monoclonal antibody (AD12) that binds to a repeated epitope on a 200 kDa adult worm excretion product present in sera from infected humans. The only formal evaluation of this assay to date was performed with sera from India. In the present study, we have evaluated the performance of the AD12 antigen assay in two laboratories with sera collected in endemic and non-endemic areas in Egypt. Antigen was detected in 57 of 59 (97%) sera from microfilaremic subjects, and in 22 of 139 asymptomatic and amicrofilaremic subjects who reside in a highly endemic area. Antigen titers were significantly correlated with microfilaria counts (r = 0.41, P less than 0.01). Filarial antigen was not detected in most sera from amicrofilaremic subjects with clinical filariasis. Comparative antigen test results obtained from laboratories in Cairo and St. Louis agreed in 170 of 173 sera tested. Filarial antigen was not detected in sera from Egyptians with no history of residence in filaria-endemic areas. Specifically, nonendemic sera from patients with other parasitic infections (schistosomiasis, fascioliasis, ascariasis, etc.) were uniformly negative in the assay. We conclude that the AD12 filarial antigen assay is sensitive and specific for W. bancrofti infection in Egypt.

Animals↗
PubMed1998

[Familial eosinophilia: contribution of parasitology to its diagnosis].

It has been observed, at the turn of the century, that members of some families showed a peripheral blood hypereosinophilia. Despite all possible investigations at that time, it was not possible to link it up with a known and well established disease. It was named "family acquired eosinophilia" by Di Guglielmo. nowadays, it has been established that some congenital defects such as the Omenn syndrome and the Wiskott-Aldrich syndrome are associated with hypereosinophilia. Eosinophilia is well known as a sign of allergy, but contaminations with helminths are an important cause of familial hypereosinophilia. In a case of familial distomatosis by Fasciola hepatica, several members of the same family were infected after eating wild cress. An epidemiological study was carried out on 22 families whose several members in a same family had an hypereosinophilia, a parasitosis was serologically diagnosed using such methods as immunoelectrophoresis, Elisa or Western blot. Among these 22 families, 10 had contracted fascioliasis and 4 who had eaten contaminated horse meat contracted trichinosis. For some other families, eosinophilia was not well documented, it concerned cysticercosis for 3 couples after eating meat of raw pork, one family with intestinal distomatosis to F. buski after consumption of aquatic plants and lastly toxocarosis, zoonosis whose contamination is acquired by geophagy but also by eating raw vegetables (salad), fruit (strawberries) and by drinking contaminated water, affected 5 families. In one family, anisakiasis with hypereosinophilia had been contracted by eating raw fish. In another one, trichostrongyloidiasis was contracted by eating raw vegetables from the family garden. The diagnosis is often oriented, for the members of a same family, who are asymptomatic or pauci-symptomatic, by the blood cell count that shows an hypereosinophilia. Furthermore, a collective outbreak of strongyloidiasis epidemic among 115 individuals by oral route is reported, showing that such a way of transmission is possible.

Adolescent↗
PubMed1998

Use of delipidized antigens of Taenia solium metacestodes in IgG-ELISA for detection of neurocysticercosis.

The development of IgG-ELISA for detecting neurocysticercosis is aimed at the routine laboratory, and requires a particular antigen preparation, an acceptable number of serum samples to be tested (both homologous and heterologous) and patients with a diversity of helminthic infections to rule out cross-reactions. This study characterizes IgG-antibodies from cases of neurocysticercosis by assaying the sera against ether-delipidized antigens (5 microg/ml) prepared from metacestodes of Taenia solium. The test had a sensitivity of 90% and a specificity of 83%. IgG-antibodies from heterologous serum samples elicited a number of false positives (25/147) from six different helminthic infections, ie paragonimiasis, echinococcosis, opisthorchiasis, ascariasis, taeniasis and fascioliasis. In additional tests to detect antibody levels to these stage-related antigens, one of three serum samples from T. solium-infected cases gave negative at OD value of 0.187 while the others yielded 0.472 and 0.576. Conversely, assays of all serum samples from neurocysticercosis cases reacted against antigens from Echinococcus granulosus cystic fluid, Paragonimus heterotremus and Opisthorchis viverrini adult worms. In comparison, the antigens from these three species yielded higher mean OD values when assayed against the corresponding infected serum samples. Furthermore, neurocysticercosis cases yielded OD values that are separate and distinct from those of paragonimiasis cases.

Adult↗
PubMedDate not supplied

[Standardization of ELISA IgM and IgA for immunodiagnosis of human trichinosis].

An ELISA test for trichinosis using as antigen a larvae soluble fraction from Trichinella spiralis was carried out for the detection of IgM and IgA specific antibodies in 45 serum samples from patients confirmed or suspected to have trichinosis by strong clinical and epidemiological evidences. All the patients had positive serology detected by precipitin test, bentonite floculation test, indirect hemagglutination test and ELISA IgG test. The cut-off value was determined using two criteria. Criterion A was determined in each plate, using three positive controls and two negative ones; the average of the negative controls and the weakest positive control, multiplied by a 1.2 factor was, considered the cut-off value. Criterion B was determined using the average plus three standard deviations from 64 apparently healthy persons serum samples. In both cases, three serum dilutions (1:10, 1:100 and 1:500) were used. The sensitivity of ELISA IgM was 100.0, 93.3 and 82.2% using serum dilutions of 1:10, 1:100 and 1:500 respectively (criterion A) and 100.0, 97.8 and 95.6% for the same dilutions (criterion B), whereas the values for ELISA IgA were: 100.0, 91.1 and 86.7% (criterion A) and 100.0, 100.0 and 91.1% (criterion B). In order to find out the specificity of ELISA IgM and ELISA IgA, additional 118 serum samples from individuals with other parasitoses, such as cysticercosis (18) hydatidosis (39), fascioliasis (12), toxocariasis (30), Chagas' disease (12) and individuals with non-specific eosinophilia (7), were also tested. ELISA IgM presented a specificity of 92.3, 93.4 and 97.3% (criterion A) and 96.2, 97.8 and 97.8% (criterion B) whereas the results for ELISA IgA were 97.8, 98.9 and 99.4% (criterion A) and 98.4% for the 1:10 and 1:100 dilutions and 100.0% for the 1:500 dilution (criterion B). The positive predictive values of ELISA IgM were 76.3, 77.8 and 88.1% (criterion A) and 86.5, 91.7 and 91.5% (criterion B) whereas the negative ones were 100.0, 98.3 and 95.7% (criterion A) and 100.0, 99.4 and 98.9% (criterion B). The positive predictive values of ELISA IgA were 91.8, 95.3 and 97.5% (criterion A) and 93.8, 93.8 and 100.0% (criterion B) whereas the negatives ones were: 100.0, 97.8 and 96.8% (criterion A) and 100.0, 100.0 and 97.8% (criterion B). The use of ELISA IgM and ELISA IgA in the immunodiagnosis of trichinosis is discussed.

Adult↗
PubMed1999

[Liver involvement in HIV infection and various parasitic diseases].

During the course of systemic infections, the liver is an organ which is frequently involved. The pathology of the human immunodeficiency virus with all its infections, opportunistic tumours and some parasitoses, such as amebiasis, kala-azar, hydatidosis, schistosomiasis and fascioliasis, evolve into hepatic compromise. This development is what we attempt to summarise.

HIV Infections↗
PubMed2000

Angiostrongyliasis: analysis of antigens of Angiostrongylus costaricensis adult worms versus IgG from infected patients with Angiostrongylus cantonensis.

The possibility of cross-reactivity was previously investigated by indirect ELISA with sera from Angiostrongylus cantonensis infections, normal controls and A. costaricensis antigen. 5 microg/ml of crude antigen from both sexes of each species reacted with diluted serum samples (1:800) of each of 20 cases of angiostrongyliasis and normal controls, and further with anti-human IgG conjugate at 1:1,000. The mean absorbance values were evaluated as follows; normal controls showed a value of 0.033 using A. costaricensis antigen lower than (0.085) A. costaricensis antigen. Both mean values of angiostrongyliasis cases were rather close (0.491) using A. costaricensis antigen and the other antigen (0.518). The present study continued with a crude antigen of 13 A. costaricensis females and males. Serum samples were analyzed; 27 sera of angiostrongyliasis, 30 negative controls and 193 cases of other parasitic infections (91 cases of nematodiasis; 45 cases of cestodiasis; 47 cases of trematodiasis and 10 cases of HIV) and 7 cases of other brain infections. This antigen was evaluated for ELISA with a concentration of 5 microg/ml, serum dilution 1:400 and anti-human IgG conjugate at 1:2,000. The test gave sensitivity and specificity at cut-off value 0.261; 92.59% and 73% respectively. The antigen was cross-reactive with 30 cases from 9 out of 10 different kinds of nematodiasis (gnathostomiasis, strongyloidiasis, ascariasis, hookworm infections, trichinosis, toxocariasis, trichuriasis, onchocercosis and Wuchereria bancrofti infections. Five cases from 3 of 6 kinds of cestodiasis (neurocysticercosis, echinococcosis and Hymenolepis nana infections) and 18 cases of 4 out of 5 kinds of trematodiasis (Paragonimus heterotremus infections, opisthorchiasis, schistosomiasis and fascioliasis). One case of other brain infections was observed. The crude antigen of A. costaricensis showed a high percentage sensitivity with serum antibodies of angiostrongyliasis cases. Low specificity of the test was observed by reactions of those serum antibodies with various kinds of antigenic molecules. This study provides baseline data for further immunodiagnosis of human angiostrongyliasis.

Angiostrongylus cantonensis↗
PubMed2002

Aspects of the maintenance of the life cycle of Fasciola hepatica in Lymnaea columella in Minas Gerais, Brazil.

Fascioliasis is a parasitic disease of domestic ruminants that occurs worldwide. The lymnaeid intermediate hosts of Fasciola hepatica include Lymnaea columella, which is widely distributed in Brazil. A colony of L. columella from Belo Horizonte, MG, was reared in our laboratory to be used in studies of the F. hepatica life cycle, the intermediate host-parasite relationship and development of an anti-helminthic vaccine. In the first experiment 1,180 snails were exposed to miracidia of F. hepatica eggs removed from the biliary tracts of cattle from the State of Rio Grande do Sul. In the second and third experiments the snails were exposed to miracidia that had emerged from F. hepatica eggs from Uruguay, maintained in rabbits. The rates of infection in the first, second and third experiments were 0, 42.1 and 0% respectively. Over 15,806 metacercariae were obtained and stored at 4 degrees C. Four rabbits weighing 1.5 kg each were infected with 32-44 metacercariae and two with 200. Three rabbits begin to eliminate eggs of the parasite in the feces from 84 days after infection onwards. The biological cycle of F. hepatica in L. columella and the rabbit was completed within 124 days.

Animals↗
PubMed2002

Fasciola immature stages sought in Lymnaea species and Biomphalaria species in the water bodies of Dakahlia Governorate.

Examination of the five different water bodies in Dakahlia governorate, revealed four species of Lymnaea. These were L. natalensis (68.4%). L. truncatula (16%), L. stagnalis (12.2%) and L. columella (3.4%). Also, two species of Biomphalaria were recovered. These were B. alexandrina (54.7%) and B. glabrata (45.3%). Examination of all these snails showed natural infection with immature stages of Fasciola sp. in 5.5% of L. natalensis (= cailliaudi), 3.1% in L. truncatula and 0.67% in B. alexandrina. The importance of these snails in dissemination and spreading of fascioliasis was discussed.

Animals↗
PubMed1997

[PARASITARY INFECTIONS OF THE LIVER]

In Perú three are the parasites which affect frequently the liver: Entamoeba histolytica, Echinococcus granulosus and hepatic Fasciola.Amebic liver abscess can start acutely or insidiously with fever and right upper quadrant abdominal pain, with or without liver or intercostal tenderness. The main complications are thoracic, peritoneal or pericardial extension (left lobe abscess). Diagnosis by images study and serology techniques are effective. Currently, Metronidazole is the treatment of choice.Hydatid liver cyst is usually asymptornatic until it produces a mass effect, allergic reactions or a cyst infection occurs as a result of rupture of the cyst to the billary ducts. Rupture to the peritoneum produces secondary seeding with protoscolices which develop into multiple intraabdominal cysts. Study techniques by images and serology methods are also very helpful for diagnosis. Surgery is the most successful treatment but there is also the possibility of medical treatment with Albendazole or a combination of both.Hepatic fascioliasis can be asymptornatic or produce fever, right upper quadrant abdominal pain and hypereosinophilia during the invasive phase, or billary tract obstruction during the chronic phase, which sometimes is only diagnosed during surgical operation. Hemobilia and biliary cirrhosis have been described as complications. During the invasive phase it is based y serological methods, and during the chronic phase diagnosis is made by detection of parasite's eggs in stools. Presently, the treatment of choice is with Triclabendazole.

Journal Article↗
PubMed2002

Bacterial-parasite interaction between Salmonella and each of Fasciola gigantica and Trichinella spiralis.

The possibility of bacterial-parasite interaction between Salmonella typhimurium and the surface of each of Fasciola gigantica metacercariae and Trichinella spiralis larvae was investigated in vitro. Two studies were carried out. In the first, S. typhimurium were incubated in vitro with the metacercariae of F. gigantica. In the second, S. typhimurium was incubated with larvae of T. spiralis. The interactions of S. typhimurium with each of F. gigantica metacercariae and T. spiralis larvae were studied by scanning electron microscope (SEM). In the first study, numerous bacilli were found adhered to the metacercariae surface. In the second study, no S. typhimurium was observed on the wall of T. spiralis larvae. The results indicated that only F. gigantica metacercariae act as a carrier for S. typhimurium with the possibility of occurrence of mixed infections with both organisms. So, both fascioliasis and salmonellosis must be treated concomitantly.

Animals↗
PubMed2002

[Immunodiagnosis of Strongyloides stercoralis infections in Chile using ELISA test].

BACKGROUND: Strongyloides stercoralis is a world wide distributed small intestinal nematode parasite. In immunocompetent individuals S stercoralis can produce asymptomatic infections or a moderate clinical picture of diarrhea, some cases become chronic. In immunocompromised patients, a disseminated disease may appear, sometimes fatal. In Chile, there is little epidemiological information about S stercoralis infections and appropriate diagnostic techniques are usually not used. AIM: To evaluate the yield of an ELISA test for the diagnosis of strongyloidiasis in Chilean patients. MATERIAL AND METHODS: Ten serum samples from patients with S stercoralis infections confirmed by a positive stool examination, 66 samples from individuals with other infections by tissue helminthes (24 toxocariasis, 15 trichinellosis, 11 hydatidosis, 12 fascioliasis and 4 cysticercosis), 13 samples from subjects with autoimmune diseases and 49 samples from apparently healthy individuals with a normal eosinophil count, were studied. ELISA antigen was prepared using a filariform larval extract obtained from a murine species of Strongyloides, maintained in laboratory animals. RESULTS: Using 0.33 optical density units as a cut off value, 9 of 10 sera of S stercoralis infected individuals, had a positive ELISA test. No cross reactions were observed with sera of patients with other helminthic infections, autoimmune diseases or in healthy individuals. Thus, specificity, positive and negative predictive values were 100%. CONCLUSIONS: The results obtained are similar with those found by other investigators. ELISA test for strongyloidiasis is a useful tool for the diagnosis of clinical cases and for seroepidemiological studies of this nematode infection in Chile.

Adult↗
PubMed2004

Evaluation of Capillaria philippinensis coproantigen in the diagnosis of infection.

In order to overcome the false negative diagnosis of infection with C. philippinesis at time of absence of eggs in stool, coproantigen prepared from stools of infected patients was evaluated serologically. This antigen was able to detect anti-Capillaria antibodies in the sera of infected cases at the same OD level produced with Capillaria crude worm antigen using indirect ELISA technique C. philippenensis coproantigen did not cross-react with sera from patients with schistosomiasis mansoni, fascioliasis or strongyloidiasis at 1:00 serum dilution. Laboratory-prepared hyperimmune sera versus crude worm antigen of C. philippinensis succeeded in capturing Capillaria antigen prepared from the stools of infected patients and did not cross react with coproantigens prepared from stool samples of cases infected with S. mansoni or Fasciola using sandwich ELISA technique.

Animals↗
PubMed2004

Alkaline phosphatase from Echinococcus granulosus metacestodes for immunodiagnosis of human cystic echinococcosis.

The work examined the use of Echinococcus granulosus alkaline phosphatase (EgAP) (extracted from hydatid cyst membranes) as an antigen for immunodiagnosis of human cystic echinococcosis (CE). It was assessed by enzyme-linked immunosorbent assay (ELISA) and Western immunoblotting (IB) for detection of serum anti-EgAP immunoglobulin (Ig)G antibody and was compared with hydatid cyst fluid (HCF). The EgAP and HCF were of sheep liver cysts origin. Sera from 30 patients with surgically confirmed CE (G. I), 30 patients with other parasitic infections (G. II), and 20 healthy controls (G. II) were examined. The mean optical density of each of anti-EgAP IgG and anti-HCF IgG antibodies in G. I was significantly higher (P < 0.01) than in each of G. II and III. The use of EgAP in ELISA showed 100% sensitivity and specificity recording significantly higher sensitivity (P < 0.05) and specificity (P < 0.01) than when using HCF in ELISA which showed 86.7% sensitivity and 84% specificity. SDS-PAGE resolution, under reducing conditions, of EgAP revealed a molecular weight of 56 KDa, while that of HCF revealed a number of antigenic bands ranged from 12 to 130 KDa. IB analysis showed that sera from CE patients recognized the EgAP 56 KDa and also one or more of HCF antigenic bands of molecular weights at 116, 63, 44, 39, 24, 20, 16 and 12 KDa. The use of EgAP in IB showed 100% sensitivity and specificity recording an insignificant difference (P > 0.05) in sensitivity and a significantly (P < 0.05) higher specificity than when using HCF in IB which showed 100% sensitivity and 90% specificity. Cross reactivity with HCF in ELISA and IB was seen with schistosomiasis mansoni, fascioliasis, hymenolepiasis nana and ascariasis. Using EgAP, there was an insignificant difference (P > 0.05) in each of the sensitivity and specificity between ELISA and IB. Using HCF, there was a significantly (P < 0.05) higher sensitivity and an insignificantly (P > 0.05) higher specificity by IB than ELISA. The implications of these results are discussed.

Adult↗
PubMed2004

Toxicological effect and biochemical alterations induced by different fractions of Euphorbia royleana latex in freshwater harmful vector snail Lymnaea acuminata.

Laboratory evaluation was made to assess the molluscicidal activity of different fractions of Euphorbia royleana (Family- Euphorbiaceae) latex obtained through sephadex gel column against freshwater snail Lymnaea (Radix) acuminata Lamarack. This snail is the vector of liver fluke, Fasciola hepatica Linnaeus and Fasciola gigantica Cobbold, which causes endemic fascioliasis in cattle and livestock. The toxic effect of the different fractions was time dependent and fifth fraction obtained through benzene: ethyl acetate (5:5) had maximum molluscicidal activity against Lymnaea acuminata. There was a significant negative correlation between LC values and exposure periods thus increase in exposure time, the LC50 value of V fraction of Euphorbia royleana latex was decreased from 14.28 mg/l (24 hr) to 9.28 mg/l (96 hr) against Lymnaea acuminata. After exposure to sub-lethal concentrations of this fraction there were significant time and dose dependent alterations observed in pyruvate, lactate levels, ALAT, AAT, AChE and cytochrome oxidase enzyme activities in different body tissues of Lymnaea acuminata. It is proposed that the fifth fraction of E. royleana latex can be used as a molluscicide for controlling the harmful snail population from aquatic ecosystem without any harm due to their reversible toxic action.

Animals↗