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Biomedical subjects

M E Scott

Publications and source records attributed to M E Scott.

At least 37 records · Page 2Linked to original sources

Energy restriction and zinc deficiency impair the functions of murine T cells and antigen-presenting cells during gastrointestinal nematode infection.

This study examined whether the impaired immune responses in zinc deficient- and/or energy-restricted mice exposed to a challenge infection of Heligmosomoides polygyrus might be associated with reduced numbers of spleen cells, altered proportions of spleen cell subpopulations and/or altered function of the T cells or antigen-presenting cells (APC). Female BALB/c mice were given free access to either a zinc-sufficient (60 mg zinc/kg diet, Zn+) or a zinc-deficient diet (0.75 mg zinc/kg diet, Zn-) or were pair-fed (PF) the zinc-sufficient diet. Significant differences in parasite burdens were observed. Worm numbers were lowest in Zn+ mice, intermediate in the PF mice and highest in the Zn- mice, showing that both zinc deficiency and energy restriction reduced protective immunity against the gastrointestinal nematode H. polygyrus. Although the absolute numbers of spleen cells were reduced in both Zn- and energy-restricted (PF) mice, neither deficiency altered the phenotypic distribution of the subpopulations of positive marker cells in the spleen. In vitro functional assays using a 1:1 ratio of APC:T cells showed that T-cell proliferation in response to parasite antigen (Ag) was impaired by a dietary effect of zinc deficiency on T cells and of energy restriction and zinc deficiency on APC function. Consequences of the nutritional deficiencies on cytokine production in response to parasite antigen were more complex: zinc deficiency reduced T-cell function [interleukin-4 and interleukin-5 (IL-4 and IL-5) production], and both nutritional deficits depressed APC functions [IL-4, IL-5, and interferon-gamma (IFN-gamma) production] and T-cell function (IFN-gamma production). Thus, this study showed that zinc deficiency and energy restriction played identifiably distinct roles in regulating host immune responses against the gastrointestinal nematode H. polygyrus.

Animals↗

Single-column purification of free recombinant proteins using a self-cleavable affinity tag derived from a protein splicing element.

A novel protein purification system has been developed which enables purification of free recombinant proteins in a single chromatographic step. The system utilizes a modified protein splicing element (intein) from Saccharomyces cerevisiae (Sce VMA intein) in conjunction with a chitin-binding domain (CBD) from Bacillus circulans as an affinity tag. The concept is based on the observation that the modified Sce VMA intein can be induced to undergo a self-cleavage reaction at its N-terminal peptide linkage by 1,4-dithiothreitol (DTT), beta-mercaptoethanol (beta-ME) or cysteine at low temperatures and over a broad pH range. A target protein is cloned in-frame with the N-terminus of the intein-CBD fusion, and the stable fusion protein is purified by adsorption onto a chitin column. The immobilized fusion protein is then induced to undergo self-cleavage under mild conditions, resulting in the release of the target protein while the intein-CBD fusion remains bound to the column. No exogenous proteolytic cleavage is needed. Furthermore, using this procedure, the purified free target protein can be specifically labeled at its C-terminus.

Affinity Labels↗

The efficacy of ivermectin against laboratory strains of Heligmosomoides polygyrus (Nematoda).

A susceptible strain of Heligmosomoides polygyrus was selected for 15 generations with increasing doses of ivermectin (0-6 mg/kg). A passage strain was developed, parallel with the ivermectin-selected strain, to control for changes due to rapid passage from mouse to mouse. The LD50s of the 8th and 15th generations of the ivermectin-selected strain were 1.5 times that of the susceptible strain. The LD50 of the passage strain at generations 8 and 15 remained similar to that of the susceptible strain. Ivermectin efficacy was lower against the LA stage than against the adult stage in the susceptible strain, the Ivermectin-selected strain and the passage strain at generation 8.

Animals↗

Life-history variation among lines isolated from a laboratory population of Heligmosomoides polygyrus bakeri.

The extent of variation in several life-history traits within a laboratory population of the parasitic nematode, Heligmosomoides polygyrus bakeri, was studied in 10 relatively inbred parasite lines isolated from a stock population and characterized in BALB/c mice after 4, 8 and 11 generations of isolation. As expected, within-line variation for most traits at generation 11 compared to generation 4 significantly decreased (P < 0.01). At each generation of characterization, variation was observed among lines for parasite establishment, rate of development in the host, rate of early egg production, per capita fecundity, short-term and long-term survival and profiles of egg production, rate of decline in egg production, life-long reproductive effort and in vitro egg hatchability. Measures of all traits, except establishment, were highly repeatable. The rate of development was higher at generation 8 compared to the stock (P < 0.0001), and regression analysis revealed that early egg production of lines increased over 11 generations of isolation (P = 0.003). These results, together with the observed decrease (P < 0.01) in total variation of most of the traits over all lines during the process of isolation, suggested an evolutionary response of the traits, probably to the rapid passage of lines every month. The rate of development subsequently decreased between generations 8 and 11 in all lines (P < 0.0001), suggesting that the random genetic drift procedure used to isolate the lines eventually exerted detrimental inbreeding effects on this trait. The evolutionary responses of life-history traits to rapid passage and inbreeding suggest a genetic basis for variation in these life-history traits.

Animals↗

Effects of three nematode anthelmintic treatment regimes on flock performance of sheep and goats under extensive management in semi-arid Kenya.

A study was undertaken in a semi-arid area of Kenya between August 1991 and June 1993 to evaluate the effects of anthelmintic treatment using ivermectin before or during the rains, on performance of mixed sheep and goat flocks, in comparison with an untreated flock. Performance parameters measured included age and weight of dams at first parturition, parturition intervals, body weights of dams and offspring, and birth weights, growth rates, and mortality rates of offspring. Among these parameters, birth weights and growth rates of offspring were found to be significantly improved by the treatment administered before the rains compared with the other two treatments. Mortality was lower in lambs and kids with high birth weights. Treatment, either before or during the rains, significantly reduced the faecal egg output and improved body weight, packed cell volume and flock fertility. Liveweight was confirmed to be a better measure of sexual maturity than age. It was further shown that lambs and kids, born of dams at their first lambing or kidding, experienced higher mortality rates than lambs and kids born of dams in their second and subsequent parturitions. Overall, treatment with ivermectin before the onset of rains was equal to or better, in terms of the performance parameters measured, than treatment during the rains, whilst treatment compared with no treatment increased performance in almost all of the parameters measured.

Albendazole↗

Zinc deficiency and energy restriction modify immune responses in mice during both primary and challenge infection with Heligmosomoides polygyrus (Nematoda).

This study characterized the consequences of zinc-sufficient (Zn+, 60 mg zinc/kg diet, ad libitum), zinc-deficient (Zn-075 mg zinc/kg diet, ad libitum) and energy-restricted (ER, 60 mg zinc/kg diet which was restricted to match food intake of Zn- mice) diets on the in vivo and in vitro immune response of BALB/c mice during both primary and challenge infection with Heligmosomoides polygyrus. In Zn+ mice, both primary and challenge infection with H. polygyrus induced not only a strong Th2 response (IgE, IgG1, eosinophilia, IL-4, IL-5, IL-10), but also elements of a TH1 response (IgG3, IFN-gamma). Zinc deficiency significantly depressed Th2-dependent antibody production during both primary and challenge infection, and reduced mitogen and antigen-induced T cell proliferation during the challenge infection. Th2 cytokine production was reduced by zinc deficiency (IL-4), energy restriction (IL-5) and by zinc deficiency possibly in combination with energy restriction (IL-10) during the primary infection whereas TH1 cytokine production (IFN-gamma) was depressed during the challenge infection by zinc deficiency, possibly together with energy restriction. Both zinc deficiency and energy restriction reduced eosinophilia with the more profound effect being exerted by zinc deficiency. Thus, both zinc deficiency and its concurrent energy restriction modify immune responses in the mice during primary and challenge infection with H. polygyrus.

Animal Nutritional Physiological Phenomena↗

High level interleukin-12 production, but diminished interferon-gamma production, by cord blood mononuclear cells.

Cell-mediated immunity (CMI) in neonates is relatively deficient when compared with adults. Defects in cytokine production and/or regulation may contribute to heightened susceptibility to infection by intracellular pathogens. The heterodimeric cytokine IL-12 is a key regulator of CMI and inducer of interferon-gamma (IFN-gamma) production. We report here that umbilical cord blood-derived mononuclear cells (MNC) are capable of producing IL-12 (p40 subunit, measured by RIA, and IL-12 p70 heterodimer, by ELISA) at levels comparable to or greater than adult peripheral blood MNC, after stimulation with heat-killed Staphylococcus aureus in 18-h cultures. As in adult MNC, S. aureus induced IL-12 p40 mRNA accumulation in cord blood MNC. IFN-gamma was also produced in the S. aureus-stimulated cultures, in an IL-12-dependent manner, but cord blood MNC produced 5-fold lower levels of IFN-gamma compared with adult MNC (p < 0.05). Preincubation with IL-10 inhibited IL-12 p40 production by cord blood and adult peripheral blood MNC in a dose-dependent fashion, whereas neutralization of endogenous IL-10 enhanced IL-12 and IFN-gamma levels. The results demonstrate that the relative CMI deficiency in neonates is not due to an intrinsic defect in the capacity of neonatal MNC to produce IL-12. The underlying factors responsible for diminished IFN-gamma production are not known, but may lie in the balance of stimulatory and inhibitory signals delivered to the IFN-gamma secreting cells along with IL-12, or may relate more to the absence of memory T cells among cord blood MNC.

Fetal Blood↗

Cloning and sequence comparison of AvaI and BsoBI restriction-modification systems.

AvaI and BsoBI restriction endonucleases are isoschizomers which recognize the symmetric sequence 5'CYCGRG3' and cleave between the first C and second Y to generate a four-base 5' extension. The AvaI restriction endonuclease gene (avaIR) and methylase gene (avaIM) were cloned into Escherichia coli by the methylase selection method. The BsoBI restriction endonuclease gene (bsoBIR) and part of the BsoBI methylase gene (bsoBIM) were cloned by the "endo-blue" method (SOS induction assay), and the remainder of bsoBIM was cloned by inverse PCR. The nucleotide sequences of the two restriction-modification (RM) systems were determined. Comparisons of the predicted amino acid sequences indicated that AvaI and BsoBI endonucleases share 55% identity, whereas the two methylases share 41% identity. Although the two systems show similarity in protein sequence, their gene organization differs. The avaIM gene precedes avaIR in the AvaI RM system, while the bsoBI R gene is located upstream of bsoBI M in the BsoBI RM system. Both AvaI and BsoBI methylases contain motifs conserved among the N4 cytosine methylases.

Amino Acid Sequence↗

Multiple infection with Plasmodium and helminths in communities of low and relatively high socio-economic status.

A study was conducted in the city of Lubumbashi, Zaire: (1) to survey parasitic infections and clinical conditions in the local children and their mothers; (2) to identify combinations of parasites and clinical conditions that commonly occurred together in individuals; and (3) to determine whether single- and/or multiple-species infections were risk determinants of the observed clinical conditions. Overall, 1100 children and mothers from three subdivisions, two of low socio-economic status (LSES) and one of relatively high socio-economic status (HSES), provided stool and blood samples and were clinically examined. Plasmodium prevalence was higher in the two LSES subdivisions than in the HSES subdivision. Prevalence and intensity of Ascaris lumbricoides infection were low in the HSES subdivision and one of the two LSES subdivisions. In contrast, prevalence and intensity of Trichuris trichiura and of hookworms were similar in all subdivisions. Plasmodium and A. lumbricoides were the most frequently found single-species infections. The combination of A. lumbricoides and Plasmodium was the most frequent double-species infection and that of A. lumbricoides, Plasmodium and T. trichiura was the most frequent triple-species infection. Significant positive associations between parasite species were detected in the HSES subdivision, and in one of the two LSES subdivisions. Because the relationships were not consistently detected, it is hypothesized that the associations are determined by environmental conditions rather than synergy between the parasites in the host. The most commonly observed clinical conditions were abdominal pain, diarrhoea, fever, and low packed-cell volume (PCV). The occurrence of each was significantly lower in the HSES subdivision than in at least one of the two LSES subdivisions. Abdominal pain and low PCV were most common in individuals presenting with only a single clinical condition, and the combination of this symptom and sign was the most commonly observed pair of conditions. Abdominal pain, low PCV and diarrhoea was the most common combination in individuals with three clinical conditions. Logistic regression revealed that hookworm infection, T. trichiura infection, young age and residence in an LSES subdivision were determinants of diarrhoea. Trichuris trichiura infection, young age and living in an LSES subdivision were risk factors for abdominal pain. Plasmodium infection and young age were risk factors for fever. LSES was the only predictor of low PCV. Infection with A. lumbricoides did not enter any of the models. No significant interactions were detected among parasites, indicating that there was no synergism or antagonism among parasites in the induced disease.

Adolescent↗

Decreased levels of complement receptor 1 (CD35) on B lymphocytes in persons with HIV infection.

Previous studies have shown that complement receptor 1 (CR1) expression on erythrocytes is decreased under several conditions including HIV infection and autoimmune diseases. The goal of this study was to determine whether expression of CR1 on peripheral blood B cells, where this receptor plays a role during immune responses, is altered in persons with HIV infection. The B cells from rheumatoid arthritis (RA) patients were also assessed since this represents a group with known complement and B cell abnormalities. The CD19+ B cells from persons with either HIV infection or RA had significantly reduced levels of CR1 when compared with control donors (75 and 72% CR1+ versus 94% CR1+ for control donors). The reduction of B cell CR1 occurred in both the percentage of B cells positive for CR1 and the levels of CR1 found on positive cells. In contrast, CR1 on monocytes was not reduced. As shown in previous studies, CR2 was also found to be reduced on B cells from the HIV-infected persons and there was extensive overlap between the B cell subsets which lacked expression of CR1 and CR2. The complement receptor-negative B cells found in HIV-infected persons were not immature or activated as defined by their lack of expression of CD10 or B7, respectively. Elevated levels of C4d, a classical complement pathway-activation product, were detected in plasma from both HIV-infected and RA patients. These studies suggest that chronic complement activation occurring in persons with HIV infection or RA can affect the complement receptor phenotype of peripheral blood B cells. Since complement receptors are involved in activation of B cells, the subset that lacks CR1 may represent cells that have encountered immune complexes and may therefore be stimulated. Additionally, the downregulation of complement receptors may have significant effects on the ability of B cells to capture and present opsonized antigens.

Antigen-Antibody Complex↗

Energy restriction and severe zinc deficiency influence growth, survival and reproduction of Heligmosomoides polygyrus (Nematoda) during primary and challenge infections in mice.

The objectives of this study were (1) to determine the impact of severe zinc deficiency on the establishment, growth, survival and reproduction of Heligmosomoides polygyrus in the laboratory mouse, during both primary and challenge infection protocols, and (2) to determine whether the observed effects resulted from zinc deficiency per se, or from the accompanying energy restriction. Three diet groups were used: zinc-sufficient (Zn+:60 mg zinc/kg diet), zinc-deficient (Zn-:0.75 mg zinc/kg diet) and energy restricted (ER:60 mg zinc/kg diet pair fed to Zn- mice). Neither Zn- nor ER influenced the establishment of the parasite during a primary infection. However, both significantly influenced the early development of the parasite. The proportion of adult worms recovered 9 days post-infection (p.i.) was highest in Zn- mice, intermediate in ER mice and lowest in Zn+ mice. Worms were also distributed more distally in the intestine of the Zn- mice and worm survival was highest in Zn- mice, intermediate in ER mice and lowest in Zn+ mice at both 4 and 5 weeks p.i. Although the length of female worms was reduced in Zn- mice, neither per capita fecundity nor egg viability was affected by zinc deficiency. Energy restriction, on the other hand, significantly reduced worm fecundity at 5 weeks post-primary infection, but had no effect on egg viability. Zinc concentration of adult H. polygyrus was similar among dietary groups. The effects of zinc deficiency and energy restriction were also investigated 4 and 5 weeks after a challenge infection. Whereas strong host resistance was evident in Zn+ and ER mice, based on comparison of worm numbers between challenged mice and primary infection controls, no evidence of resistance was detected in Zn- mice. As in the primary infection, female worms were shorter in Zn- mice than in ER and Zn+ mice, and energy restriction but not zinc deficiency significantly affected per capita fecundity. However, in contrast to the primary infection, ER mice had elevated rather than reduced fecundity. This study demonstrates a complex interaction between H. polygyrus and zinc and energy restriction, and highlights the importance of controlling for reduced food intake in nutrition-infection studies.

Animals↗

Ascaris lumbricoides infection and environmental risk factors in an urban African setting.

Identification of appropriate strategies for controlling gastro-intestinal nematodes in communities depends, in part, on an understanding of the conditions that increase risk of exposure to infective stages. The present study was conducted in Lubumbashi, Zaire. The objectives were to identify features of the environment and living conditions that were significant predictors of Ascaris lumbricoides infection, and to determine whether the same predictors were important in populations living in subdivisions of lower (LSES) and higher (HSES) socio-economic status. Forty-two households from each of three subdivisions (two LSES and one HSES) were selected at random. Mothers were interviewed, observations on the environment around the home were recorded, and single stool samples, collected from all children and mothers, were examined for nematode eggs. Maternal education was a significant predictor of A. lumbricoides intensity in both LSES and HSES subdivisions. Factors related to poor sanitation (open defecation and high number of people using the same toilet) were important in the LSES subdivisions but not in the HSES subdivision. In contrast, the ratio of relatives to direct family members per household was a significant predictor of high intensity of infection in the HSES subdivision, but not in the LSES subdivisions, indicating that relatives and live-in visitors contribute to Ascaris transmission in the HSES population.

Adolescent↗

Evaluation of non-invasive blood pressure measurement by the Finapres method at rest and during dynamic exercise in subjects with cardiovascular insufficiency.

The accuracy and precision of the Finapres in recording rest and exercise blood pressure compared with the intra-arterial (aortic and branchial) and random-zero sphygmomanometer methods was assessed in 84 ischaemic patients in three different studies. Firstly, comparison at rest with the aortic intra-arterial pressure in 50 ischaemic patients demonstrated that the Finapres systolic (136.5 +/- 21.1 vs. 129.3 +/- 19.0 mmHg; p < 0.001) and mean (92.4 +/- 13.4 vs. 90.7 +/- 11.4 mmHg; p < 0.001) arterial pressures were higher and diastolic pressures lower (70.4 +/- 11.5 vs. 71.5 +/- 9.8 mmHg; p < 0.001). The reproducibility of the Finapres and invasive method was similar for systolic (4.6% vs. 4.0%), diastolic (2.8% vs. 2.7%) and mean (3.3% vs. 3.0%) blood pressures. Second, in seven subjects studied twice at rest and during 4 min supine bicycle exercise, the exercise increase in blood pressure was greater on the Finapres compared with the brachial intra-arterial pressure (systolic +10.2 +/- 6.3 vs. +3.6 +/- 9.8 mmHg; diastolic +9.6 +/- 11.1 vs. +0.2 +/- 2.1 mmHg; p = 0.02 for each); however, at steady-state the peak/trough differences in pressure between the methods were similar. Thirdly, compared under rest conditions, to random zero sphygmomanometer (RZO), the Finapres systolic pressure was higher (6.8 +/- 3.5 mmHg) and diastolic pressure lower (-6.0 +/- 1.9 mmHg). During upright bicycle exercise, the difference between the Finapres and RZO in systolic blood pressure increased at each level of exercise (+14.3 +/- 4.2, +17.9 +/- 4.0 and +22.2 +/- 4.1 mmHg respectively at each exercise stage: p < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Genetic variability of the beta-tubulin genes in benzimidazole-susceptible and -resistant strains of Haemonchus contortus.

Benzimidazole anthelmintics are the most common chemotherapeutic agents used to remove intestinal helminths from farm animals. The development of drug resistance within helminth populations is wide-spread and can render these drugs essentially useless. The mechanism of benzimidazole resistance appears to be common to many species ranging from fungi to nematodes and involves alterations in the genes encoding beta-tubulin. During the selection process resulting in resistance, there must be quantitative changes in the population gene pool. Knowledge of these changes would indicate the mechanisms underlying the spread of resistance in the population, which in turn could be used to design more effective drug administration strategies. To this end we have identified allelic variation at two beta-tubulin genes in Haemonchus contortus using restriction map analysis of individual adults. Extremely high levels of variation were identified at both loci within a susceptible strain. In two independently derived benzimidazole resistant strains, allele frequencies at both loci were significantly different from the susceptible strain but not from each other. The same alleles at both loci, in both resistant strains, were favored by selection with benzimidazoles, suggesting that both loci are involved in determining benzimidazole resistance. These data confirm that changes in allele frequency, rather than novel genetic rearrangements induced by exposure to the drug, explain the changes associated with benzimidazole resistance. These results also show that any DNA based test for the development of benzimidazole resistance must take into account the frequency of alleles present in the population and not simply test for the presence or absence of specific allelic types.

Alleles↗

Zinc deficiency impairs T cell function in mice with primary infection of Heligmosomoides polygyrus (Nematoda).

This study was designed to determine whether severe zinc deficiency would prolong the course of a primary Heligmosomoides polygyrus infection in mice, and whether this could be related to impaired T cell function. Female BALB/c mice were fed a zinc-sufficient (Zn+; 60 mg/kg), a zinc-deficient (Zn-; 0.75 mg/kg) or an energy restricted (PF; 60 mg zinc/kg) diet. After four weeks, some mice in each dietary group were given a primary infection with 100 larvae; nutritional, parasitological and immunological parameters were assayed over the following five weeks. Liver zinc concentrations were significantly reduced in Zn- mice compared with Zn+ mice. In certain cases, PF mice also had reduced liver zinc concentrations, showing the negative effects of restricted food intake on zinc status. Zinc deficiency prolonged the course of a primary infection, with the effects being most evident five weeks post-infection when Zn+ mice had only 40% as many worms as Zn- mice. Parasite infection induced strong immunological responses in Zn+ mice in contrast to Zn- mice. The reduced production of IL-4 and IFN-gamma, the reduced peripheral eosinophilia and reduced serum levels of IgE and IgG1 in Zn- mice were attributed to the zinc deficiency, whereas the reduced delayed type hypersensitivity response to parasite antigen and reduced production of IL-5 were in certain instances attributed to reduced energy intake rather than zinc deficiency. These results show that zinc deficiency significantly impairs functions normally attributed to both Th1 and Th2 cell populations, and that these alterations are associated with elevated worm numbers in zinc-deficient mice.

Animals↗

Inhibition of human factor VIIIa by anti-A2 subunit antibodies.

Human inhibitory alloantibodies and autoantibodies to Factor VIII (FVIII) are usually directed toward the A2 and/or C2 domains of the FVIII molecule. Anti-C2 antibodies block the binding of FVIII to phospholipid, but the mechanism of action of anti-A2 antibodies is not known. We investigated the properties of a patient autoantibody, RC, and a monoclonal antibody, 413, that bind to the region which contains the epitopes of all anti-A2 alloantibodies or autoantibodies studied to date. mAb 413 and RC were noncompetitive inhibitors of a model intrinsic Factor X activation complex (intrinsic FXase) consisting of Factor IXa, activated FVIII (FVIIIa), and synthetic phospholipid vesicles, since they decreased the Vmax of intrinsic FXase by > 95% at saturating concentrations without altering the Km. This indicates that RC and mAb 413 either block the binding of FVIIIa to FIXa or phospholipid or interfere with the catalytic function of fully assembled intrinsic FXase, but they do not inhibit the binding of the substrate Factor X. mAb 413 did not inhibit the increase in fluorescence anisotropy that results from the binding of Factor VIIIa to fluorescein-5-maleimidyl-D-phenylalanyl-prolyl-arginyl-FIXa (Fl-M-FPR-FIXa) on phospholipid vesicles in the absence of Factor X, indicating it does not inhibit assembly of intrinsic FXase. Addition of Factor X to Fl-M-FPR-FIXa, FVIIIa, and phospholipid vesicles produced a further increase in fluorescence anisotropy and a decrease in fluorescence intensity. This effect was blocked completely by mAb 413. We conclude that anti-A2 antibodies inhibit FVIIIa function by blocking the conversion of intrinsic FXase/FX complex to the transition state, rather than by interfering with formation of the ground state Michaelis complex.

Animals↗

Influence of storage and examination methods on the recovery and size of metacercariae of Cyathocotyle bushiensis and Sphaeridiotrema pseudoglobulus (Digenea).

The recovery of metacercariae of Cyathocotyle bushiensis (Digenea: Cyathocotylidae) and Sphaeridiotrema pseudoglobulus (Digenea: Psilostomidae) from Bithynia tentaculata (Gastropoda: Prosobranchia) subjected to 3 methods of storage (70% alcohol, freezing, and fresh) and 2 methods of examination (crushing or artificial digestion) was evaluated. Prevalence and abundance of fully developed metacercariae of C. bushiensis and metacercariae of S. pseudoglobulus did not significantly vary as a result of storage or examination methods; however, digestion of frozen and fresh snails resulted in a significantly lower prevalence and abundance of not fully developed metacercariae of C. bushiensis. Furthermore, freezing or storing snails in alcohol may result in misclassification of the metacercariae of C. bushiensis. Both methods of storage and examination also affected the size of the metacercariae of both digeneans. Although digested snails were easier to examine, there appeared to be no overall gain in time by using digestion instead of crushing.

Analysis of Variance↗