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K D Murrell

Publications and source records attributed to K D Murrell.

At least 91 records · Page 5Linked to original sources

Evaluation of an antigenic fraction of Taenia hydatigena metacestode cyst fluid for immunodiagnosis of bovine cysticercosis.

An antigenic fraction (ThFAS) isolated from Taenia hydatigena metacestode cyst fluid was used in an ELISA to detect antibodies to T saginata in experimentally and naturally infected cattle. In 10 calves given 1,000 to 100,000 T saginata eggs (20% to 60% viability), IgG and IgM antibodies were detected in all the calves by post-inoculation week 3. Immunoglobulin G antibody values remained increased until calves were slaughtered at post-inoculation weeks 13 to 26. Six naturally infected calves (determined by postmortem examination) were considered positive, using the ELISA. Shared antigens were demonstrated between ThFAS and T saginata and T crassiceps; there were no shared antigens between ThFAS and Haemonchus contortus or Fasciola hepatica. Specific lectin binding to ThFAS indicated the presence of glycoconjugates. Immunoblot analysis indicated that a low molecular weight polypeptide (10,000 Mr) bears the immunodiagnostic antigen.

Animals↗

Epidemiologic findings on a swine farm with enzootic toxoplasmosis.

A farm in Illinois had swine with enzootic Toxoplasma gondii infections. Ninety-five of 99 pigs had antibody against T gondii by the modified agglutination test; modified agglutination test titers were less than 1:10 (4 pigs), 1:10 (15 pigs), 1:20 (12 pigs), 1:40 (10 pigs), 1:80 (20 pigs), 1:160 (11 pigs), 1:320 (12 pigs), 1:640 (9 pigs), and greater than or equal to 1:1,280 (6 pigs). To trace the route of infection, Toxoplasma-free pigs were introduced into the farm and were evaluated serologically at various intervals. Analysis of data derived from these tracer pigs indicated that cannibalism was a major source of T gondii infection in the pigs.

Agglutination Tests↗

Trichinosis in a herd of swine: cannibalism as a major mode of transmission.

In a herd of approximately 1,000 hogs, evaluation of muscle specimens collected at various intervals during a 12-year period (1973 to 1985) indicated continuous transmission of Trichinella spiralis. The farm's rat population and the incidence of trichinosis in the rats was high during 1974, but diminished markedly by 1978. In January 1984, a longitudinal investigation, using tracer pigs, was performed to determine whether rodents and/or other wild animals were involved in transmission of T spiralis on this farm. Tracer pigs exposed to rodents and wild animals did not become infected with T spiralis. The rodent population on the farm was small and none of the rodents trapped and examined were found to be infected. Hog cannibalism also was evaluated as a mode of T spiralis transmission. Results of the investigation indicated that hog cannibalism was the mode of transmission for trichinosis in the herd.

Animals↗

Distribution of Toxoplasma gondii tissue cysts in commercial cuts of pork.

Distribution of Toxoplasma gondii cysts in tissues used for human consumption was studied in 4 naturally infected pigs (acquired infections) and in 2 experimentally infected pigs. On the day of euthanasia, pigs with acquired toxoplasmosis had serum antibody titers of 160 to 1,280 against T gondii (modified agglutination test). Experimentally infected pigs were inoculated with 10,000 oocysts of the GT-1 strain of T gondii and were euthanatized 267 or 357 days after inoculation. On the day of euthanasia, the experimentally infected pigs had serum antibody titers of 16 and 32 against T gondii. Fifty- to 100-g specimens from each of 6 cuts of meat commercially used to prepare Boston butt, arm picnic, ham, tenderloin, spareribs, and bacon and from the tongue, heart, brain, liver, kidneys, and diaphragm were collected from each pig and were digested in acid pepsin solution. Washed sediment from the digest of each specimen was inoculated SC into mice (6 mice/organ). Toxoplasma gondii was isolated from tissues of the 4 pigs with acquired toxoplasmosis; T gondii was isolated from the arm picnic, Boston butt, and tongue of 3 pigs, from the ham, tenderloin, spareribs, diaphragm and heart of 2 pigs, and from the bacon and brain of 1 pig. Toxoplasma gondii was not isolated from the kidneys of the 4 pigs with acquired toxoplasmosis. Toxoplasma gondii was isolated from the arm picnic, Boston butt, ham, spareribs, bacon, tongue, diaphragm, heart, and brain of the 2 experimentally infected pigs, and from the kidneys of one of the experimentally infected pigs.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Trichinella spiralis: antifecundity and antinewborn larvae immunity in swine.

The development of antifecundity and antinewborn larvae immunity in swine infected with Trichinella spiralis was investigated. In primary infections, adult female worm fecundity dropped sharply after 3 weeks, although adults could be recovered from the small intestine for at least 7 weeks after infection. In challenge infections of pigs infected previously, adult female worm fecundity was depressed up to 51% and the adults were expelled within 3 weeks. Since immune pigs are almost completely resistant to the secondary establishment of muscle larvae, this suggested the existence of immune effector mechanisms also acting on the newborn larvae. This was supported by observations, using an indirect fluorescent antibody assay, that pig antibody bound to the surface of the newborn larvae. Passive transfer of immune pig serum resulted in a large reduction in muscle larvae burden in both infected pig and rat recipients. Adult female worm fecundity in such immune serum recipients was reduced only by 20% and worm survival in the intestine was unaffected. These results indicate that immunity to the newborn larvae, in addition to antifecundity effects, are responsible for the high levels of acquired resistance to T. spiralis in swine.

Age Factors↗

Trichinella spiralis in the black bear (Ursus americanus) of Pennsylvania: distribution, prevalence and intensity of infection.

Samples of tongue or diaphragm from 2,056 black bears harvested in Pennsylvania during the 1981-1983 hunting seasons were examined for larvae of Trichinella spiralis by peptic digestion. Sixteen males and 21 females were infected. The overall prevalence of infection was 1.8%. Infected animals were distributed widely throughout the range of the bear in Pennsylvania. In samples from infected bears, the geometric mean density of muscle larvae was 7.8 per g of tissue (LPG). There were neither sex- nor age-related differences in prevalence or intensity of infection. Virtually all bears harvested in Pennsylvania are consumed as food, which often is shared widely among hunters, their friends and relatives. Furthermore, high densities of larvae occurred in some bears (i.e., 300, 348, 465, 512, 555, and 912 LPG). Thus, a basis for potential, single-source outbreaks of severe human trichinosis exists.

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Conservation of diagnostic antigen epitopes among biologically diverse isolates of Trichinella spiralis.

Crude and immunoaffinity-purified excretory-secretory antigens derived from a domestic pig isolate of Trichinella spiralis were used in an enzyme-linked immunosorbent assay to test serum from mice infected with 25 different pig and wild animal isolates of T. spiralis sspp. All of the sera were found positive by ELISA using either of the antigen preparations, indicating all isolates shared certain antigen epitopes. Excretory-secretory antigens were prepared from 3 distinct isolates of T. spiralis sspp.--Trichinella spiralis spiralis (pig isolate), Trichinella spiralis nativa (polar bear isolate), and Trichinella spiralis pseudospiralis--and compared by electrophoresis and monoclonal antibody binding. While protein profiles varied among the isolates, a monoclonal antibody recognizing a major immunodiagnostic antigen epitope bound all 3 antigen preparations. However, this antigen epitope occurred on different molecular weight excretory-secretory proteins from the different isolates.

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Field evaluation of the enzyme-linked immunosorbent assay for swine trichinosis: efficacy of the excretory-secretory antigen.

A field evaluation of an enzyme-linked immunosorbent assay (ELISA) for swine trichinosis was done with sera obtained from 5 herds experiencing ongoing transmission of Trichinella spiralis. Epizootiologic studies conducted on these herds offered an opportunity to evaluate the accuracy of an ELISA, using larval T spiralis excretory-secretory antigens. Sera from 162 infected pigs and 143 serum samples from noninfected pigs originating from the same farms were tested. The infection status of the pigs was determined by digestion of diaphragm or tongue muscle samples. Two criteria were established to classify the ELISA optical density (OD) readings: Criterion I stated that an OD greater than or equal to 5 times the mean OD of several normal swine sera pools was positive; criterion II stated that a OD greater than or equal to 4 times the normal sera values was positive. The results obtained did not reveal obvious serologic variations among infected herds located in the 4 states involved. Overall, the test detected 93% (criterion I) and 96% (criterion II) of infected pigs. The majority of false-negative sera was from hogs that had less than 5 larvae/g of muscle; 1 hog had 73.8 larvae/g of diaphragm muscle. The false-positive rates were 8% for criterion I and 9% for criterion II. The actual rate for these false-positive samples may have been overestimated, because generally, only small tissue samples (0.4 to 10 g) were digested; larger sample sizes might have altered the results. The relevance of this qualification is that these pigs originated from herds with prevalence rates greater than 50%. Other factors that may account for occasional false-positive sera or false-negative sera in the swine trichinosis ELISA are discussed.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Inoculation of pigs against Trichinella spiralis, using larval excretory-secretory antigens.

Pigs were inoculated with Trichinella spiralis excretory-secretory products derived from short-term in vitro maintenance of infective muscle larvae. Intraperitoneal administration of excretory-secretory products in Freund's complete adjuvant or aluminum hydroxide induced moderate, but variable, degrees of immunity to challenge exposure in a nondose-dependent manner; IM administration of products in Freund's incomplete adjuvant was less successful. Inoculated pigs harbored fewer adult worms, and the fecundity of female worms (numbers of newborn larvae shed in vitro) recovered after challenge exposure was significantly lower (alpha = 0.05) than the fecundity of females recovered from control pigs. The degree of resistance in inoculated pigs was directly related to serotiter against excretory-secretory antigens, as determined in an enzyme-linked immunosorbent assay.

Aluminum Hydroxide↗

Effect of irradiation on the viability of Toxoplasma gondii cysts in tissues of mice and pigs.

Muscles from tongue, heart, and limbs of 14 pigs inoculated orally with Toxoplasma gondii oocysts were irradiated with 10, 20, 25, and 30 krad of gamma (cesium-137 and cobalt-60) irradiation. Viability of T gondii cysts was assayed by feeding porcine muscles to T gondii-free cats and/or by inoculation of sediment from acid-pepsin digested porcine muscle into mice. Cats fed 500-g samples of muscles irradiated with up to 20 krad shed T gondii oocysts. Cats fed muscles irradiated with 25 or 30 krad did not shed oocysts. Mice were inoculated with 8 isolates of T gondii, and tissue cysts in their brains irradiated with up to 40 krad were infective to mice; however, there was a 10,000-fold reduction in the viability of organisms in tissue cysts irradiated with 40 krad, compared with that in nonirradiated cysts. At 50 krad of gamma irradiation, there were no detectable infective organisms in infected mouse brains.

Animals↗

Strongyloides ransomi: proteolytic enzymes from larvae.

The filariform larvae of Strongyloides ransomi can infect their hosts by penetration through skin. In this report, homogenates of these organisms were prepared and their proteolytic enzymes examined. Homogenates prepared in 0.2 M citrate, pH 4.0, contain two thiol-dependent proteinases with molecular weights of approximately 32,000 and 28,000. These proteinases have an acidic pH optimum and show substrate preferences and inhibitor susceptibilities similar to the vertebrate acidic cysteinyl proteinases. Homogenates prepared in 0.1 M Tris, pH 7.5, contain multiple proteolytic enzymes, active against both Azocoll and synthetic substrates. These enzymes do not require thiols for activity and they have an alkaline pH optimum. The enzymes are inhibited by both chelating agents and heavy metals, but not by serine-proteinase inhibitors. Extracts prepared in 0.1 M Tris-HCl, pH 7.5, contain endogenous proteinase inhibitors.

Animals↗

Trichinella spiralis: acquired immunity in swine.

The ability of domestic pigs to develop protective immunity to Trichinella spiralis in response to inoculation with different doses of muscle larvae was assessed. Adult worms developing from the inoculations of 112, 500, and 10,000 larvae were expelled from the intestine about 6 weeks after inoculation. Inoculation with 25,000 larvae, however, resulted in more rapid intestinal worms expulsion, indicating that gut expulsion is dose dependent. Secondary expulsion also tended to be dependent upon primary infection level. Pigs initially inoculated with 500 to 10,000 larvae expelled the challenge infection of adult worms after 22 to 25 days; in contrast, infection by inoculation of only 112 larvae failed to induce significant enchanced gut expulsion of the challenge infection intestinal worms. However, all primary infection levels, including inoculation with 112 larvae, induced nearly absolute resistance to the muscle establishment of larvae from challenge adult worms. The fecundity of female worms recovered from immune pigs was reduced 75% in comparison to controls. These results show that, in contrast to some host species, very rapid gut expulsion does not occur in domestic swine. Yet, immune responses at the gut level are important, perhaps responsible for much of the inhibition reflected as reduction in the establishment of muscle larvae.

Animals↗

A potential diagnostic reagent for bovine cysticercosis.

A fraction of larval Taenia hydatigena cyst fluid was shown to have high sensitivity and specificity in the enzyme-linked immunosorbent assay (ELISA) for the detection of bovine antibodies to the heterologous parasite Taenia saginata. This antigenically active lipoprotein fraction was isolated by ultracentrifugal density flotation using either ammonium sulfate (specific gravity = 1.231 g per ml) or NaCl/KBr (specific gravity = 1.225 g per ml), followed by ion-exchange chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) indicated that this fraction was composed of high molecular weight (65,000 to 77,000 Mr) and low molecular weight (9,500 to 16,000 Mr) proteins. Electrophoresis under non-denaturing conditions in either acrylamide (5%) or agarose (1%) resulted in 1 major diffuse band staining for both protein and lipid. The high and low molecular weight proteins observed on SDS-PAGE under reducing conditions could not be resolved by gel filtration chromatography and emerged as a single lipoprotein peak. This T. hydatigena cyst fluid fraction appears promising as a diagnostic reagent in the ELISA for bovine cysticercosis.

Animals↗

Swine trichinosis in New England slaughterhouses.

Diaphragms of 5,315 slaughter hogs from the New England states were examined for Trichinella spiralis infection between June 22 and Dec 22, 1983. Thirty-nine hogs (0.73%) were infected. The infections were light, none exceeding 30 larvae/g of tissue, with an arithmetic mean of 4.7 larvae/g. Infected hogs originated from 5 of 6 New England states. The widespread occurrence of porcine trichinosis in New England indicates that the small or part-time hog farming operations frequently found in this region may be especially vulnerable to the introduction and maintenance of the parasite.

Abattoirs↗

Trichinella spiralis: immunization of rats with an antigen fraction enriched for allergenicity.

Trichinella spiralis whole muscle larval extract was fractionated by gel filtration and anion-exchange chromatography, and the protein fractions were assayed for allergenicity by a footpad-swelling test in mice; IgE antibody levels in rats immunized with the fractions were determined by passive cutaneous anaphylaxis test in rats. By these methods, an allergenic fraction from T. spiralis was isolated. The fraction, F1-b, was shown to be monodisperse by analysis with SDS-PAGE, IEP, and isoelectric focusing, indicating that it is a single protein moiety with a molecular weight of approximately 45,000 and a pI of approximately 5.1. The Schiff-periodate test showed F1-b to be a glycoprotein. Rats immunized with F1-b had significantly fewer intestinal worms than did nonimmunized controls at 24 hr and 7 days after oral challenge with T. spiralis larvae.

Animals↗

Immunization of swine against Trichinella spiralis.

Swine were immunized with partially purified stichosome antigens derived from Trichinella spiralis muscle larvae. In 3 trials, 500 to 600 micrograms of the solubilized particle component (S3) induced moderate levels of resistance to challenge inoculation; the percent reduction in larvae per gram of muscle ranged from 43 to 55. The effect of immunization with S3 on pig intestinal expulsion of adult worms was determined in one experiment and the results indicated that S3 immunization was only weakly effective in enhancing gut expulsion. These results suggest that immunization with S3 induces a protective immune response in the pig that may be directed either against the migrating newborn larvae or against adult worm fecundity.

Animals↗