The use of zeolites as slow release anthelmintic carriers

This work examines the ability of commercial zeolite Y to act as a slow release agent for a number of anthelmintic drugs. Administration to rats, dosed with Nippostrongylus brasiliensis, of pyrantel and/or fenbendazole and pigs, dosed with Ascaris and Oesophagostomum, of dichlorvos (DDVP) loaded onto zeolite Y was more successful in killing adult worms than administration of the pure drug alone. The zeolite Y was used as supplied for initial studies and then later dealuminated for further studies. The drug loadings were monitored by thermal analysis and the loaded zeolites were used in several field trials. The results indicate that zeolite Y is a suitable vehicle for the slow release of some anthelmintics. The slow release of drug from the zeolite matrix improved its efficacy.     

Angiostrongylus cantonensis: Rejection of pulmonary arterial transfers of lung stage worms

Experimental transfer of the lung stage worms of Angiostrongylus cantonensis was performed between permissive hosts (rats) and between permissive (rat) and nonpermissive hosts (guinea pigs and rabbits). These worms from rats were rejected when implanted into nonpermissive hosts. Unexpectedly, similar worms did not survive well even in permissive hosts; the majority of recipient rats did not have first-stage larvae (L₁) in their stools and, even when positive for L₁, the number of the larvae shed was few. These findings contrast with the successful pulmonary arterial transfer of younger, intracranial-stage worms. It was shown that differences in rat strain between donor and recipient had no significant effect on the subsequent worm survival in recipient hosts. The alteration of maintaining conditions of the intrapulmonary worms, prior to transfer, in terms of temperature, media, and maintaining period, also showed no profound effect on the subsequent worm survival. The kinetics of precipitating and reaginic antibody levels in rats implanted with the intrapulmonary worms were analogous to those in rats with intracranial-stage worms. The findings indicate that some qualitative differences may exist between the worms obtained from two different sites.     

Strongyloides ratti: reversibility of immune damage to adult worms

Transplantation experiments were conducted to assess the reversibility or irreversibility of the damage sustained by Strongyloides ratti during infections in the rat host. Worms of different ages from primary and secondary infections were recovered from their original hosts and transplanted surgically into naive rats. The size and fecundity of normal (Days 6–11 postinfection) worms were maintained after transfer. Damaged worms from primary infection (Days 22–26) showed complete recovery of size and fecundity within 10 days of transfer; damaged worms from a secondary infection (Days 6–7) also showed functional recovery but to a lesser extent. The ultrastructural changes observed mainly in the intestine of damaged worms from primary infections, prior to their transfer, were, however, only partially ameliorated following transplantation into new naive hosts; there was no complete return to structural normality. On the other hand, second infection worms did show almost complete ultrastructural recovery. The course of a transplanted infection established with either damaged or normal worms was similar to infections established percutaneously. Increase in the size of transplanted infections from 100 to 250 worms per recipient did not alter the dynamics of the host/parasite relationship. There was no evidence of adaptation in S. ratti and damaged worms, when transplanted into naive rats, were as successful as normal worms in protecting the host against a subcutaneous larval infection. The implications of this work on the present understanding of the phenomenon of autoinfection in experimental rodent strongyloidiasis are discussed.     

Survival of Onchocerca volvulus in nodules implanted in immunodeficient rodents.

Onchocerca volvulus is an obligate human parasite, and its study has been difficult due to an inability to maintain it outside the human host. We report the successful transplantation of onchocercomata containing living adult O. volvulus worms into immunodeficient C.B.-17.scid/scid (scid) mice or athymic rnu/rnu (nude) rats. Living, motile worms containing viable microfilariae were present in onchocercomata recovered from scid mice or nude rats for up to 20 wk, establishing a novel animal model for future investigation of O. volvulus.     

Host-parasite relationship of Angiostrongylus cantonensis. 1. Intracranial transplantation into various hosts.

Various stages of Angiostrongylus cantonensis recovered from the brain of experimentally infected mice were transplanted intracranially into rats. Third and fourth-stage worms recovered 2--7 days postinfection were able to develop normally after transplantation into recipient rats. The fifth-stage worm obtained 14--15 days postinfection would enter the brain tissue of rats but died shortly afterwards. However, the same stage of worms recovered from rats, after a similar transplantation, were found to develop normally in the recipient animal. Young fifth-stage worms, from the subarachnoid space of rats, which were ready for the pulmonary migration were also transplanted into rabbits but the worms failed to reach the lungs. In the control rat-to-rat transplantation, the worms successfully completed the pulmonary migration. The morphogenesis and initial growth rate of A. cantonensis were similar in both mice and rats but in the former host the worms started to grow at a markedly slower rate after the last moult and gradually degenerated.     

负载金属离子的5A 沸石的体外抗菌实验

目的:探讨负载金属离子的5A沸石的体外抗菌作用。方法:选用金黄色葡萄球菌、铜绿假单孢菌、白色念珠菌,利用倍比稀释法对5A沸石组、磺胺嘧啶银组及载不同浓度Ag+、Zn2+5A沸石共15组进行了体外抗菌试验研究,确定最佳抗菌效果离子负载方案。结果:三种细菌的MIC,双金属离子负载在抗菌上具有协同载Ag+5A沸石分别达到了125μg/ml~500μg/ml、31.25μg/ml~500μg/ml、250μg/ml~500μg/ml;附载Zn2+5A沸石分别达到了12.5mg/ml~25mg/ml、6.25mg/ml~50mg/ml、25mg/ml;负载双离子5A沸石分别为250μg/ml~500μg/ml、62.5μg/ml~500μg/ml、500μg/ml。结论:5A沸石负载金属离子后均具有抗菌作用,且抗菌作用与附载金属离子的量正相关;负载相同质量Ag+的5A沸石较附载Zn2+者具有更强的抗菌作用;双金属离子负载在抗菌上具有协同作用;2%Ag++8%Zn2+与2%Ag++10%Zn2+及4%载银组与阳性对照组无显著性差异。     

Nippostrongylus brasiliensis: reversibility of reduced-energy status associated with the course of expulsion from the small intestine in rats

Gastrointestinal nematodes require energy for active establishment in the gut against intestinal flow and peristaltic motion. In this study we employed CellTiter-Glo Luminescent Cell Viability Assay to measure the ATP value of individual adult Nippostrongylus brasiliensis during the course of immune-mediated expulsion from the small intestine in rats. The ATP values of adult worms taken from the lumen of the distal small intestine were lower than worms collected from the lumen of the proximal small intestine. Moreover, values from worms in the lumen of the proximal small intestine were lower than those from worms in the mucosa, the preferred site of adult N. brasiliensis. The reduction of ATP values in worms from each region was observed not only at expulsion phase, but also at established phases of the infection suggesting that energy metabolism of the parasites is independent of host immune response. When adult worms with low ATP values on day 12 post-infection were implanted surgically into the small intestine of na?ve rats, the worms re-established in recipients and completely restored the ATP values. Short in vitro culture of adult worms under low oxygen tension resulted in low ATP value in the worms. These results suggested that adult worms were dislodged from their preferred site by intact energy metabolism activity.     

负载金属离子的5A沸石的体外抗菌实验

目的：探讨负载金属离子的5A沸石的体外抗菌作用。方法：选用金黄色葡萄球菌、铜绿假单孢菌、白色念珠菌,利用倍比稀释法对5A沸石组、磺胺嘧啶银组及载不同浓度Ag＋、Zn2＋5A沸石共15组进行了体外抗菌试验研究,确定最佳抗菌效果离子负载方案。结果：三种细菌的MIC,双金属离子负载在抗菌上具有协同载Ag＋5A沸石分别达到了125μg/ml~500μg/ml、31.25μg/ml~500μg/ml、250μg/ml~500μg/ml;附载Zn2＋5A沸石分别达到了12.5mg/ml~25mg/ml、6.25mg/ml~50mg/ml、25mg/ml;负载双离子5A沸石分别为250μg/ml~500μg/ml、62.5μg/ml~500μg/ml、500μg/ml。结论：5A沸石负载金属离子后均具有抗菌作用,且抗菌作用与附载金属离子的量正相关;负载相同质量Ag＋的5A沸石较附载Zn2＋者具有更强的抗菌作用;双金属离子负载在抗菌上具有协同作用;2%Ag＋＋8%Zn2＋与2%Ag＋＋10%Zn2＋及4%载银组与阳性对照组无显著性差异。     

Schistosoma mansoni: relationship between parasite age and time of spontaneous elimination from the rat

Laboratory rats infected with Schistosoma mansoni invariably reject the majority of parasites between the fourth and sixth week after infection (self-cure). This investigation was designed to determine whether the timing of rejection is dictated by the rat or by the parasite. Schistosomes were perfused from rats infected 2, 3, or 4 weeks previously and were transferred into the mesenteric veins of normal rats. Recipient animals were perfused at weekly intervals after transfer, and the timing of worm elimination was determined in recipients. It was found that 2-week-old worms were rejected 2 weeks after transfer, 3-week-old worms 1 week after transfer, and 4-week-old worms immediately after transfer. Schistosomes perfused from mice or hamsters and transferred into rats showed the same pattern of worm elimination. It is concluded that at the fourth week of normal schistosome development there is a critical event which makes virtually impossible any further survival of the parasite in laboratory rats.     

The reversibility of intestinal immune expulsion effects on adult Nippostrongylus brasiliensis

An attempt has been made to study the extent and nature of the damage occurring in adult Nippostrongylus brasiliensis undergoing immune expulsion from the rat. It was found that worms are not killed nor irreparably damaged when being rejected. On transfer into naive second recipient rats the rate of re-establishment of worms previously incubated in immune rat recipients for 4-17 hr was high (68-69%) and comparable to that shown by worms from normal recipient rats (48-56%). Similarly, worms taken on days 10, 11, and 12 of a primary infection, already passed to the distal half of the small intestine due to immune expulsion effects, on transfer into naive recipient rats re-established themselves well (rates varying from 62 to 80%) compared to those harvested from their normal habitat in the proximal half of the small intestine (rates varying from 44 to 87%). Worm damage is associated with decreased motility and impaired locomotion capacity. The phenomenon of mucosal trapping occurs during expulsion, but merely to the extent of some 30% of the worm population. It is suggested that in principle, worms subjected to immune expulsion are in a state of acute, transient metabolic crisis. The present results support the enteroallergic indirect mechanism for worm rejection.     

Nippostrongylus brasiliensis infections in mice: the immunological basis of worm expulsion.

Nippostrongylus brasiliensis infections in mice were terminated more rapidly than in rats and immunologically induced damage occurred earlier. Like rats, mice expelled damaged worms more rapidly than normal worms. Recipients of cells from the spleen or mesenteric node of immune mice expelled their worms by day 8 of the infection. Recipients of cells alone or antiserum alone did not expel their worms by day 5 but mice given both cells and antiserum expelled their worms by this stage of the infection. Damaged worms were expelled more rapidly than normal worms from mice given immune cells, This work indicated that antibodies and cells collaborate to expel N. brasiliensis from mice as has been shown to occur in rats.     

Bactericidal activity of Ag-zeolite mediated by reactive oxygen species under aerated conditions

The bactericidal activity induced by the introduction of silver ions into zeolite was studied. Escherichia coli was used as the test microorganism. Silver ions were loaded into zeolite by the ion-exchange method. Silver-loaded zeolite was demonstrated the strong bactericidal activity. Dissolved oxygen was an essential factor for the occurrence of the bactericidal activity because the activity was observed only under aerated condition. Superoxide anions, hydrogen peroxide, hydroxyl radicals and singlet oxygen were formed. Scavengers of these each reactive oxygen species (ROS) inhibited the bactericidal activity. This means that all ROS contributed to the activity.     

Comparative morphology and ecology of two species of Litomosoides (Nematoda: Filarioidea) of rodents in Florida, with a key to the species of Litomosoides Chandler, 1931

From April 1970 to August 1971, 235 cotton rats (Sigmodon hispidus) and 281 rice rats (Oryzomys palustris) were trapped in salt and fresh water marshes in north-central Florida and examined for natural infections of filarial worms, Litomosoides spp. Cotton rats from both types of marshes were infected (28–44 per cent prevalence), whereas only rice rats from the salt marsh were infected (56 per cent prevalence). Older animals were more commonly infected than younger ones. In cotton rats the worms were located in the pleural cavity, whereas in rice rats the worms were located primarily in the abdominal cavity. Filarial worms from rice rats were transmitted experimentally to laboratory-reared rice rats and cotton rats, but worms from cotton rats were transmitted only to cotton rats. Morphological studies on adult forms and microfilariae indicated that the worms in rice rats were distinct from those in cotton rats and are therefore described as Litomosoides scotti sp. n. The cotton rat filariids were referable to Litomosoides carinii (Travassos, 1919) Vaz, 1934. L. scotti differs from L. carinii in the ratio of the spicules, in the shape of the distal end of the right spicule and in having shorter microfilariae.     

Acquired immunity in rats against Angiostrongylus cantonensis infection

Acquired immunity against Angiostrongylus cantonensis was induced by immunizing rats with somatic antigens from fifth-stage larvae and adult worms and live third-stage larvae. Rats immunized twice had significantly fewer worms than rats immunized three times. Fewer worms were recovered from rats immunized with 200 live third-stage larvae than from any other groups. Rats immunized with somatic antigens had higher enzyme-linked immunosorbent assay (ELISA) antibody levels than rats immunized with live larvae. Rats immunized with live third-stage larvae of Angiostrongylus cantonensis were more strongly protected against challenge infections (62-92%) than rats immunized with antigens extracted from fifth-stage larvae (0-30%) and adult worms (11-24%).     

Schistosoma mansoni: immunization of cynomolgus monkeys by injection of irradiated schistosomula.

Although the immunization of primates with irradiated schistosome cercariae has been demonstrated, no success has been reported by injection with the irradiated schistosomule stage. The present investigation was designed to test whether cynomolgus monkeys could be protectively immunized with ⁶⁰Co-irradiated Schistosoma mansoni schistosomula. Monkeys injected once with 10⁴ irradiated schistosomula (50 krad at 4 krad/min) had 52% fewer challenge worms than the control group at necropsy. Four immunizations did not induce a higher level of resistance. At 50 days post-challenge, the immunized monkeys excreted 80% fewer eggs than did the control animals. An attempt to enhance irradiated schistosomule-induced protection with tetramisole · HCl was unsuccessful.     

Influence of host strain and helminth isolate on the first phase of the relationship between rats and Moniliformis moniliformis (Acanthocephala)

Eleven trials, involving 440 rats bred from 3 laboratory strains and worms from 4 isolates of Moniliformis moniliformis, were carried out with each rat receiving an oral dose of 15 cystacanths. The results showed that the infectivity of the cystacanths was not affected by their age (range 55-194 days) or by their density per cockroach during development (16.1-88.6 cystacanths per cockroach). The numbers of worms per rat recovered at 35 days postinfection (p.i.) were shown to be related to rat strain, with highly inbred strains (PVG and F344) being more supportive of numbers of worms than an outbred Wistar strain. There was no evidence to suggest that the sex of the rats had any influence on the numbers of worms recovered at 35 days p.i. Evidence was obtained to suggest that smaller (younger) rats are likely to support more worms on average than larger (older) rats. There was no evidence of any relationship between worm weight and numbers of worms present per rat on day 35 p.i. Generally, rat strain had little effect on the dry weight (growth) of male M. moniliformis, in contrast to observations made for female worms. The greatest range of worm weights was observed from the recent isolate of the worm (1982) as compared with the well established isolate (1956) and the rats that supported most worms differed from those that harbored the largest worms. Rat sex was not observed to be associated with worm weight. The frequency distributions of numbers of M. moniliformis per rat were not described readily by the negative binomial distribution.     

Completion of Schistosoma mansoni life cycle in thyroidectomized rats and effects of thyroid hormone replacement therapy

The survival and maturation of Schistosoma mansoni worms was analyzed in normal, thyroidectomized (Thyrox), and hormone-restored Thyrox rats. Restoration therapy was conducted with both T3 and T4; weight gain of treated rats was monitored to assess hormone-replacement efficacy. Worm yields, lengths, and sex-ratios were compared. Egg yields and the capacity of miracidia to hatch from eggs were also analyzed. The results of these studies support the conclusion that the sequence of steps leading to completion of the S. mansoni life cycle are operational in Thyrox rats. Hormone-treated Thyrox rats are restored to the nonpermissive status, although the worms isolated from these rats still differ in certain respects when compared to worms isolated from normal rats.     

Endemic echinostome infections of candidate hosts

Wild Lymnaea tomentosa snails, recovered from Lake Wanaka, New Zealand, were established in the laboratory. Wild snails, naturally infected with echinostomes, provided metacercariae for infection of laboratory maintained snails. Metacercarial cysts from wild and laboratory snails were then used to attempt infection of definitive host candidates. Laboratory snails provided convenient packaging of known numbers of cysts. Metacercariae excysted in the small intestines of ducklings to mature in 6 days. Worms were expelled as they became gravid. Attempts to establish infections in experimental hosts other than ducklings were not successful. No worms were recovered from mice, white rats, guinea pigs, hamsters or immunosuppressed white rats.     

A sand-zeolite culture system for simulating plant acquisition of potassium from soils

A sand-zeolite culture system simulating plant acquisition of K from soils has been developed. The system, a mixture of synthetic zeolite IE-96 and coarse-sand, provides K concentrations comparable to soils depending on the K concentration ratios of solutions loaded onto the cation-exchange sites of the zeolite and the ionic strengths of the nutrient solutions supplied during the period of plant growth. Tomato (Lycopersicon esculentum Mill.) strain 203, PI 124163, was used in this study. An increase in the amount of the loaded zeolite per culture pot did not shift solution phase K activity ratios (AR^k) of the culture system but did result in a linear increase of plant dry matter accumulation, indicating that K bioavailability is diffusion-limited in the sand-zeolite culture system as in soils.     

Factors influencing the fecundity of Moniliformis moniliformis (Acanthocephala): constant diet and varied dose

Aspects of the reproductive performance of Moniliformis moniliformis were investigated in rats allowed to feed ad libitum on a purified diet containing 1% (w/w) fructose as an energy source for the worms. The rats were infected with either 10, 20, 40 or 80 cystacanths each with the intention of investigating density-dependent effects on worm fecundity. The establishment of the worms in the gut was independent of dose, but survival, growth and reproductive performance generally were shown to be related to the infective dose given to the rats. The effects could not be related to the absolute numbers of worms present in the small intestine at post-mortem examination. In general, some unidentified regulatory process appeared to operate to create severe density-dependence in survival so that surviving parasites were not present in numbers expected to generate competition. Attainment of sexual maturity, growth and the production of mature eggs by worms from rats given doses of 80 cystacanths each were delayed compared with worms from rats given the other doses, but eventually the performance of the high-dose worms caught up. Worms attached more anteriorly in the small intestine grew bigger and produced more mature eggs. Possible mechanisms responsible for the observed effects are discussed.