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[
Parasitology,
1979]
Infective larvae of homogonic Strongyloides ratti grown in faecal culture with 32P or 75Se acquired a significant amount of radioactivity which was firmly attached to them. Heating removed most of the 32P but left 75Se in place. Subcutaneous injection of virgin and nursing mother rats with living and heat-killed radioactive larvae resulted in a pattern of labelling in the small intestine of injected animals and, in the case of 75Se, those of suckling pups, which can only be explained if labelled worms follow the natural migratory routes. The use of this tool in migratory studies is discussed, with precautions to allow for flaws in the technique.
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[
Parasitology,
1978]
Eight days after mother rats were injected with 4000 infective larvae of Strongyloides ratti at different stages of lactation the numbers of adult worms in their intestines were uniformly low (less than 3% of the dose) compared with unmated controls (mean = 25%). Those in their litters varied from 12% on day 5 to a maximum of 47% on day 17 post partum. These data, which do not correlate with lactational performance, imply that parasite movements in lactating rats are controlled by qualitative, not quantitative, consequences of humoral events. The numbers of worms in litters are concluded to be the result of the interaction of dynamic determinants of larval routes in the mother and changes in the suitability of the neonatal gut as an environment for worm development. The timing of events leading to milk-borne infection is defined. Injected larvae were closely synchronized in their movements, which were completed in 36 h. Larvae experimentally diverted into the mother's tissues during her first lactation were not available for the infection of a second litter.
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[
Parasitology,
1976]
The numbers of mature worms which developed in young rats after their mothers were injected with 4000 L3 late in lactation were 1% (Nippostrongylus brasiliensis) and 24% (Strongyloides ratti) of the dose administered. The low value for N. brasiliensis validates the conclusion that milk is a real and important vehicle for infection in S. ratti since possible errors from skin invasion of the young would have been common to both species. The level of mature S. ratti infection in lactating mothers in this experiment was negligible, 97-99% of the adult worms appearing in the offspring. These results may indicate that the milk route is possible with N. brasiliensis even though it is quantitatively insignificant.
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[
Parasitology,
1982]
A prediction of the hypothesis of Wilson (1977, 1980 a, b) to account for larval migration of homogonic Strongyloides ratti in the host is that the pattern of invasion of the mammary gland of a lactating rat will be quantitatively similar on both sides and independent of the point of entry into the body. Twenty-one suckled mother rats in 6 experiments in which live 75Se-labelled 3rd-stage homogonic larvae were injected under the skin of the upper flank had an overall distribution of label 30 h post-injection, as a percentage of the initial dose, in the quadrants, I (rear, injection side, II (rear, opposite injection side), III (front, injection side) and IV (front, opposite injection side) of the mammary gland as follows: 27.4%, 1.27%, 1.98% and 1.24%. Quantitative changes in mammary label between 30 and 48 h post-injection using live larvae, differences between mothers and virgins, and results after injection of heat-killed labelled larvae, confirm that the pattern is representative of the behaviour of normal (unlabelled) worms when injected. The theory is therefore disproved. The findings are put forward as the first quantitative evidence for major lymphatic involvement in migration of a skin-penetrating roundworm. They need confirmation in similar experiments in which worms are allowed to penetrate the skin naturally. The role of isotope-labelled larvae versus traditional methods of estimating parasite content of host tissue is discussed.
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[
J Helminthol,
1986]
Subcutaneous injection of the larvae is the almost universally adopted means of initiating experimental infections of skin-invading roundworms but, so far, the possibility that this procedure introduces artefacts of one kind or another has not been critically studied. Experiments described in this paper were used to compare the effect of (a) injection and (b) skin application, of a small, precisely counted ('exact') dose of larvae. Results with two strains of S. ratti showed that the same proportion of the dose developed to adults in the intestines of rats irrespective of the method. With the same exact dose technique it has been shown that milk-borne infection of the pups of lactating rats is not an artefact produced by injection. Large doses (mean 4000) of larvae of the homogonic strain of S. ratti carrying a radioactive label of 75Se were tracked in their migration to the mammary gland following injection or skin application at two different sites on the right-hand side of nursing mother rats. The broad conclusion of earlier work in this laboratory using injection, that larvae move by a local route and not a systemic one, was supported by the results. The detailed distribution of the label and of unlabelled worms of the heterogonic strain in families was, however, different for the two methods, indicating that subtle variations in pathway can be brought about by the use of injection. If migration involves the lymphatic system, then the interpretation of immunological experiments in terms of lymphatic anatomy must take account of such procedural effects. The extent to which these results contribute to theories of migration in Strongyloides ratti is discussed.
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[
Parasitology,
1978]
There were no differences in mean intestinal worm burdens 8 days after subcutaneous injection of 4000 infective larvae of Strongyloides ratti into rats in dioestrus, pro-oestrus, oestrus and metoestrus. Thus, changes in the hormonal environment of the migrating larvae dependent on the oestrous cycle did not alter the worms' destination or affect their potential for development. In particular, the results are prima facie evidence that prolactin is not, on its own, responsible for the re-orientation of larvae in the tissues of nursing mothers. Other sources of variability in experimental S. ratti infections are analysed and the 'exact dose' technique offered as a corrective for some procedural errors.
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[
Parasitology,
1982]
Unsuckled mother rats given a 1 h suckling stimulus 3 h after subcutaneous injection of an exact dose of homogonic Strongyloides ratti allow fewer worms to develop in their intestines by day 9 than nulliparous rats (Wilson & Simpson, 1981). This effect is studied in more detail in terms of the length of time between weaning and stimulus (W leads to S) and injection and stimulus (I leads to S). It was observable with a W leads to S of 30 h but this and a period of 5 h were less effective than 24 h. With W leads to S constant at 24 h, significantly more worms developed in mothers when I leads to S was 24 h compared to 3 h and 10 h (P less than 0.005). The data, combined with those from nulliparous controls, are presented as a measure of the change with time of numbers of larvae in that compartment of the system which gives access to the stimulated mammary gland. It is argued that the particular compartment is the local lymph node draining the injection site and that the kinetics deduced are applicable to migration in the rat in general.
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[
Parasitology,
1976]
Subcutaneous injection of nursing mothers from day 16 to day 20 post partum with infective larvae of Strongyloides ratti or Nippostrongylus brasiliensis does not result in the development of worms in the litters if 1 h is allowed between injection and resumed nursing and suckling is terminated 24 h later. Thus the low numbers of N. brasiliensis (1% of the dose) which develop in litters after 24 h, 4 day or 5 days suckling when mothers are injected and returned to their young immediately, represent skin invasion and not milk-borne infection. Taking precautions consistent with the foregoing, S. ratti equivalent to 28%, 45%, 45% and 48% of the dose were shown to be transmitted in the milk to suckling rats of 4 mothers injected with 4000 L 3 on day 18 post partum and 72 h before weaning. One mother of the same batch failed to transmit worms (1% of the dose in the litter) and the take in all 5 mothers was insignificant (max. = 3% of the dose versus an average of 21% in controls). Large numbers of S. ratti were subsequently found in the intestines of mothers whose litters were weaned immediately after (21%) or 6 h after (16%) injection, whereas very few (less than 1%) developed in mothers deprived of their offspring 24 h after injection. Dynamic, rather than static, determinants of larval routes inside the host present the only logical basis for an explanation of these facts.
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[
Parasitology,
1990]
Rigorous proofs applicable to the routes of migration of Strongyloides ratti and Nippostrongylus brasiliensis skin-penetrating juveniles inside the rat are extended. By applying the inequality principle (Tindall & Wilson, 1988) it was confirmed with a probability of error of 1 in 10(10) that N. brasiliensis larvae applied to the skin passed through the lungs on their way to the intestine. Taking the analysis further, migrating larvae of S. ratti or N. brasiliensis were extracted from the nose or lungs, respectively, of donor rats and transferred to recipients by stomach tube to assay their ability to colonize the intestine. Results showed that (a) changes undergone by each parasite in its proven, specific transit site were essential before larvae could establish in the intestines of recipients, (b) these changes could be monitored by morphological criteria, and [corrected] (c) these changes were not completed until larvae had been in the nose or lung for a significant period. It follows from (c) that anywhere in the body of the host, termed a 'nursery', that supports a substantial amount of this mandatory development must be detectable by the conventional procedure of sampling at autopsy. Conversely, absence of parasites judged by sampling at autopsy is positive proof that a site is not a nursery when sampling is timed in relation to reliable estimates of overall kinetics (Tindall & Wilson, 1990), and with control information on the efficiency of sampling. Comparative data from sampling at autopsy using the same extraction techniques for both species met these criteria: they demonstrated that no part of the head of the rat was a nursery for N. brasiliensis, and that the lung did not serve in this capacity for S. ratti.(ABSTRACT TRUNCATED AT 250 WORDS)
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[
Parasitology,
1988]
The first rigorous proof applicable to the migration pathway of an infective juvenile macroparasite inside its host is presented. Third-stage larvae of a homogonic strain of Strongyloides ratti applied in exact doses of less than 20 to the skin of the flank of young rats were recovered 16-40 h later in the naso-frontal part of the head. The peak proportion of the dose (po) recovered between 20 and 25 h in this site had a mean value of 0.316 +/- 0.021 in 48 animals. In 40 other rats infected simultaneously the mean proportion of the dose (pf) that reached the small intestine was at least 0.837 +/- 0.013. Proof resides in verification of the inequality po + pf greater than 1. With appropriate statistical tests the excess of the sum of the means of these two proportions over unity is shown to have a probability of occurring by chance of 1 in 3.5 x 10(6). Thus it is effectively certain that the naso-frontal portion of the head is part of at least one pathway taken by this parasite on its way from the skin to the intestine of its host. By suitable protection of the infection site it was confirmed that migration to the head was achieved by an internal route and not as a result of grooming. Larvae were recovered from the cranium in the same rats over the period 15-40 h, but the peak proportion of the dose occurred at 20 h, and po + pf less than 1 in this location. Whether the cranium is also part of the pathway is therefore still undecided. The significance of this novel analysis in the general context of in-host migration of infective stages is discussed and it is concluded, following its application to data sets from other authors, that the only cases in which proof can be demonstrated are the anterior skull of the rat for S. ratti (present data) and the lung of the same host for Nippostrongylus brasiliensis (Twohy, 1956).