Anthelmintic profile of methyl 5(6)-(4-methylpiperidin-1-yl) carbonylbenzimidazole-2-carbamate in experimental helminthiases

Biological evaluation of methyl 5(6)-(4-methylpiperidin-1-yl) carbonylbenzimidazole-2-carbamate against Ancylostoma ceylanicum, Nippostrongylus brasiliensis, Syphacia obvelata, Hymenolepis nana, H. diminuta and Cysticercus fasciolaris in experimental animals is reported. The compound (mg/kg) causes 100% elimination of A. ceylanicum (25 x 1), N. brasiliensis (100 x 1), S. obvelata (50 x 1), H. nana (250 x 3) and C. fasciolaris (50 x 10). It was also effective against the developing larvae (L3, L4 and L5) of A. ceylanicum at a single oral dose of 100 mg/kg. Another study indicated that the compound elicits 100% response within 32 hr of drug administration. The drug is well tolerated and LD50 is greater than 4500 mg/kg.     

A review of the endoparasites of wild House Mice Mus domesticus

A tabular checklist of the parasites of wild House Mice Mus domesticus is presented, with data on prevalence rates and geographic location. Parasites most often recorded are: Syphacia obvelata, Taenia taeniformis, Aspiculuris tetraptera, Hymenolepis diminuta and Trichuris maris. The near-global distribution of House Mice and their ability to survive in a wide range of conditions render House Mouse parasites ideal for the study of environmental influences on parasites and transmission of parasites to new hosts.     

Enhancement of FK-506 activities by ganglioside (GM3) in vivo

Influence of FK-506 and ganglioside (OM3) on acquired immunity to Hymenolepis nana reinfection was examined in BALB/c mice. Treatment of mice with FK-506 abolished the acquired immunity to challenge infection with eggs of H. nana when the agent was injected inraperitoneally at daily doses of 9.0 to 11.0 mg/kg (but not 8.5 mg/kg) on days -1, 0, 1, 2 and 3 relative to the challenge. Intravenous injection of GM3 at a daily dose of 10.0 mg/kg could not produce an immunosuppressive effect on the acquired immunity. Combination treatment with 8.5 mg/kg/day FK-506 and 10.0 mg/kg/day of GM3 inhibited the acquired immunity. These results suggest that GM3 will be a good candidate for a clinical supplementary immunosuppressive agent.     

Efficacy of selamectin against mites (Myobia musculi, Mycoptes musculinus and Radfordia ensifera) and nematodes (Aspiculuris tetraptera and Syphacia obvelata) in mice

The effects of selamectin were studied in mice naturally infected with the mites Myobia musculi, Myoceptes musculinus and Radfordia ensifera and with the oxyurid nematodes Aspiculuris tetraptera and Syphacia obvelata. The mice were divided into three treated and three control groups (n=9). Selamectin in the range 10-12.4 mg/kg was applied topically to the skin in a single spot at the base of the neck in front of the scapulae. The mice of treated and control groups were necropsied on the 4th, 7th and 21st day after the treatment. While selamectin was 100% effective in removing M. musculi, M. musculinus and R. ensifera by the seventh day, its effect against S. obvelata and A. tetraptera was 36.7% and 49.2%, respectively on the 21st day.     

Anthelmintic effects of bithionol, paromomycin sulphate, flubendazole and mebendazole on mature and immature Hymenolepis nana in mice

The anthelmintic effects of anti-tapeworm drugs, bithionol, paromomycin sulphate, flubendazole and mebendazole on immature and mature Hymenolepis nana in mice were compared. Immature worms were not affected by paromomycin sulphate or flubendazole administered for 12 consecutive days (days one to 12 after infection) at 100 mg/kg/day but 48% and 100% of H. nana were eliminated from mice by bithionol and mebendazole respectively, at the same dosage regimen. Bithionol, paromomycin sulphate, flubendazole and mebendazole given at 100 mg/kg/day for five consecutive days (days 12 to 16 after infection) eliminated 32%, 29%, 36% and 100% of mature worms respectively. 10 and 20 mg of mebendazole/kg/day for five consecutive days (days 12 to 16 after infection) had little effect on mature worms whereas 50 and 100 mg/kg/day for the same period eliminated 99% and 100% of mature worms, respectively. ED50 of mebendazole in the elimination of mature H. nana was 14 or 15 mg/kg/day for five days from the reduction in dry weight or in number of worms recovered respectively. The effects of mebendazole given 2 to 4 days, 8 to 10 days or 13 to 15 days after infection at 100 mg/kg/day were compared. Very low, if any, activity of the drug given 2 to 4 days after infection was seen, whereas the drug given 8 to 10 days or 13 to 15 days after infection eliminated 84% and 86% of H. nana respectively.     

The Hymenolepis diminuta-golden hamster (Mesocricetus auratus) model for the evaluation of gastrointestinal anticestode activity

A novel laboratory anticestode assay was developed using Hymenolepis diminuta in the hamster. The commercial anticestode compounds, praziquantel, bunamidine, and niclosamide were active against patent infections of Hymenolepis diminuta in golden hamsters (Mesocricetus auratus) when given orally at 3.125, 100, and 200 mg/kg, respectively. The gastrointestinal nematode anthelmintics, cambendazole and mebendazole, were active at 50 mg/kg. Rafoxanide (fasciolicide) was active at 25 mg/kg, the lowest level tested. The coccidiostat, nicarbazin, was active at experimental levels (800 mg/kg and up). The anthelmintic-ectoparasiticide (endectocide), ivermectin, was inactive against the tapeworm at 0.5 mg/kg, as expected.     

Efficacy of methyl 5(6)-(alpha-hydroxyphenylmethyl) benzimidazole-2-carbamate, a metabolite of mebendazole, against developing forms of experimental helminth parasites.

Methyl 5(6)-(alpha-hydroxyphenylmethyl) benzimidazole-2- carbamate, a metabolite of mebendazole, was evaluated against metamorphic forms of Ancylostoma ceylanicum in hamsters, Nippostrongylus brasiliensis in rats and cysticercoids of Hymenolepis nana in grain beetles. The test compound offered better action than mebendazole except against H. nana cysticercoids where the activity of the compound and mebendazole was comparable, but was inferior to the standard cestodicidal drug, praziquantel. The results suggest that the action was better by ip route compared to per os route of drug administration.     

Effect of chronic ivermectin treatment on GABA receptor function in ethanol withdrawal-seizure prone and resistant mice

Ivermectin, a potent, effective anthelmintic, is easy to administer, has a broad spectrum of action and a wide safety margin. However, no testing has been done in hosts genetically selected for seizure susceptibility which may be more sensitive to the effects of ivermectin than other animals. This was done in the present experiments with seizure prone and seizure resistant mice infested with Syphacia obvelata (pinworm). These subjects were treated daily with oral ivermectin in their drinking water every other week for six weeks, for a total of 21 days. The treatment cleared the mice of the pinworm infestation, but did not alter the seizure susceptibility or binding parameters of [3H]flunitrazepam in either of the selected lines.     

Mediation of isoproterenol-induced thirst in rats by beta2-adrenergic receptors.

Isoproterenol-induced thirst in rats has been attributed to the activation of beta-adrenergic receptors. Since these receptors can be further differentiated pharmacologically into beta1 and beta2 types, experiments were performed using several beta-adrenergic agonists and antagonists to determine the receptor type initiating the isoproterenol-induced thirst. The beta1- and beta2-adrenergic antagonist, d,l-propranolol (1 mg/kg, ip), blocked the increase in water intake usually accompanying acute subcutaneous administration of isoproterenol (25 microgram/kg) to female rats. Since l-propranolol is known to stabilize membranes and to possess anesthetic-like properties, d-propranolol was also used. This isomer has little beta-adrenergic-blocking activity but possesses anesthetic-like activity. Administration of d-propranolol (1 mg/kg, ip) failed to affect the drinking response to acute administration of isoproterenol (25 microgram/kg). Practolol (125 mg/kg), a beta1-adrenergic antagonist with little anesthetic properties, also had no effect on water intake of isoproterenol-treated rats. Butoxamine, a selective beta2-adrenergic antagonist, attenuated the drinking response to isoproterenol. Salbutamol (150 microgram/kg), a beta2-adrenergic agonist, mimicked the effect of isoproterenol on water intake. These results are consistent with the suggestion that beta2-adrenergic receptors mediate the isoproterenol-induced thirst in rats.     

Carcinogenesis with the insecticide rotenone

Rotenone is an insecticide which has been used extensively for a long time, and is now widely used in the U.S.A. and other industrialized countries. Rotenone is used mainly as an agricultural insecticide, household garden insecticide and water plant pesticide. Through these uses, rotenone may now be reaching the human male and female in these countries in substantial amounts, carried by fresh or cooked vegetables, consumed fish and drinking water. A review of the existing published reports and unpublished official documents dealing with the capacity of rotenone to induce neoplastic, paraneoplastic and preneoplastic lesions in the rat is presented here. It is strongly suggested that rotenone is carcinogenic to the rat (at doses from 2 to 25 parts per million continuously in food, or from 0.8 to 2.5 mg/kg weight for 1 to 4 months by oral administration), above all, when the rats receive deficient diets, especially those poor in riboflavin. Rotenone carcinogenesis, among other things, seems to exhibit a peculiar dose response pattern, which could be explained by its possible hormonal mechanism of action. Further studies to assess definitively the role of rotenone as a possible environmental carcinogen are proposed as being highly necessary.     

Long-term population dynamics of pinworms (Syphacia obvelata and Aspiculuris tetraptera) in mice

The population dynamics of concurrent infections of Syphacia obvelata and Aspiculuris tetraptera (Nematoda) in laboratory mice were investigated under conditions of constant re-exposure over periods of 56 and 115 days. The results indicate that A. tetraptera burdens equilibrate at a higher level than S. obvelata burdens and that both parasites become aggregated in the mouse population. Parasite burdens were higher following short-term (7 day) exposure of uninfected tracer mice to both parasites when compared with parasite burdens attained over long-term exposure, indicating probable development of immunity. A significant positive correlation was detected between numbers of immature S. obvelata and immature A. tetraptera for both experimental and tracer mice.     

Aspiculuris tetraptera in wild Mus musculus. The prevalence of infection in male and female mice.

A survey was carried out of the levels of infection with Aspiculuris tetraptera and Syphacia obvelata in a wild house mouse population living in the Charles Clore Small Mammals Pavilion at the London Zoo in Regent's Park. The extent of infection with A. tetraptera is analysed according to the sex of the host. It is shown that the prevalence of infection was greater in male than in female mice and frequency distribution studies suggest that this is not only because fewer female mice become infected but also because females resist larvae more effectively than do males.     

Blast cell activity in mice infected with Hymenolepis nana, H. diminuta and Trichinella spiralis: in vivo uptake of 125IUdR in lymphoid tissues and gut

The kinetics of the lymphoblast response in mice during the course of a primary infection with Hymenolepis nana was measured by the in vivo uptake of 125IUdR. The response was most marked in tissues local to the site of infection, involving the nodes draining the small intestine but not other areas, e.g., inguinal lymph nodes. A close correlation between these responses and the course of infection was observed. Uptake of 125IUdR was greatest in the mesenteric lymph node (MLN) but the peak reached in this organ was later than that in Peyer's patches (PP), small intestine (SI) and spleen (S). The increase in lymphoblast activity of the MLN was similar with Trichinella spiralis; no significant blast cell response to infection with H. diminuta was found till day 9 after injection, the results being similar to those obtained when H. nana infections were established using cysticercoids rather than eggs. It has been shown that the increase in lymphoblast activity was closely correlated with the presence of cells which are most effective in adoptive transfer immunity. A dose-dependent effect was detected in blast cell activity of MLN in different infection levels with T. spiralis and H. nana.     

Population distribution and zoonotic potential of gastrointestinal helminths of wild rats Rattus rattus and R. norvegicus from Jamaica

The population distribution and zoonotic potential of gastrointestinal helminths in a naturally infected population of wild rats (Rattus rattus and Rattus norvegicus) in Jamaica are described. One hundred and thirty (29.7%) of 437 rats captured in the study were infected: 104 (35%) of 297 R. rattus compared with 26 (18.6%) of 140 R. norvegicus. Nine species of gastrointestinal helminths were recovered: Raillietina sp. (0.2%), Trichuris sp. (0.2%), Rictularia sp. (0.7%), Syphacia obvelata (1.1%), Strongyloides ratti (1.4%), Hymenolepis diminuta (3.8%), Protospirura muricola (4.3%), Moniliformis moniliformis (11.2%), and Nippostrongylus brasiliensis (14.2%). In a logistic model, the single risk factor identified for both M. moniliformis and P. muricola was R. rattus, compared with R. norvegicus (OR = 8.369 and 9.714, respectively). In comparison, the risk factor predicted for infection with N. brasiliensis was the northeastern section of Jamaica (OR = 11.000) compared with western Jamaica. Rictularia sp. represents a new geographic distribution record for the Caribbean region. Hymenolepis diminuta, M. moniliformis, Raillietina sp., and Rictularia sp. are potentially zoonotic, but only human infection with H. diminuta has been previously reported in the Caribbean.     

Artificial Infections of Pneumocystis carinii in the SCID Mouse and their Use in the In Vivo Evaluation of Antipneumocystis Drugs

A model for the in vivo evaluation of antipneumocystis drugs has been developed in SCID mice infected intratracheally with cryopreserved mouse-derived Pneumocystis carinii. The development of a highly reproducible fatal P. carinii pneumonia occured within 10 weeks (mean survival time ± SEM = 72.2 ± 1.2 days). Continuous administration of dexamethasone (2 mg/liter in the drinking water) exacerbated the rate of onset of severe P. carinii pneumonia (mean survival time ± SEM = 63 ± 1.3 days) in SCID mice. The number of cysts per g of lung homogenate (homogenate counts) were maximal with an inoculum of 20,000 cysts at 6 weeks post infection. Homogenate counts correlated with infection scores (graded assessments of immunofluorescent cysts on lung impression smears) suggesting that infection scoring accurately and rapidly reflects the severity of P. carinii pneumonia in SCID mice. These studies led to the development of a drug screening protocol in which Pneumocystis-free female SCID mice (20–25 g) were started on dexamethasone 7 days prior to IT inoculation with a single dose of 20,000 cysts. Drugs were evaluated either for: a) prophylaxis (continuously from day 1 post infection) or b) treatment (from day 21 post infection) until day 42 post infection, when all mice were killed and infection scores determined. Co-trimoxazole (at 250 mg sulfamethoxazole + 50 mg trimethoprim/kg/day) given in the drinking water was found to be highly effective in both the prophylaxis and treatment of mouse P. carinii pneumonia. Co-trimoxazole remained very effective in the prophylaxis P. carinii pneumonia in the SCID mouse at 125 mg sulfamethoxazole + 25 mg trimethoprim/kg/day p.o. and showed some enhancement of efficacy over sulfamethoxazole alone at 125 mg/kg/day p.o., suggesting limited synergy between sulfamethoxazole and trimethoprim. The results presented provide confirmation of the usefulness and predictability of the model.     

Chromium bioaccumulation in rice grown in contaminated soil and irrigated mine wastewater--a case study at South Kaliapani chromite mine area, Orissa, India

The level of chromium (Cr) contamination in soils and irrigated mine wastewater at South Kaliapani chromite mine region of Orissa, (India) were investigated. Chromium bioaccumulation in rice plants (Oryza sativa L. cv. Khandagiri) irrigated with Cr+6 contaminated mine wastewater was analyzed along with its attenuation from mine wastewater. The levels of Cr+6 in irrigated mine wastewaters in successive rice grown plots were analyzed on 75 days and 100 days after transplantation of seedlings. Total chromium content in different parts of rice plants and soil samples from different plots was analyzed during harvesting stage (125 days after transplantation). Cr accumulation was significantly high in surface soils (0-20 cm) with a mean value of 11,170 mg kg(-1), but it decreased significantly after the crop harvest. About 70% to 90% reduction of Cr+6 levels was observed in irrigated mine wastewater when passed through successive rice plots. High bio-concentration of Cr in leaves with values ranging from 125-498 mg kg(-1) as compared to stem (25-400 mg kg(-1)) and grain (5-23 mg kg(-1)) was noticed. The reduction of Cr+6 levels is related to plant age, high biomass and area of water passage and was attributed to rhizofiltration technique.     

Effects of Aspiculuris tetraptera dn Syphacia obvelata on exploratory behavior of an inbred mouse strain

Sixty C57BL/6 male mice were treated with an anthelmintic for 2 weeks to free them of pinworms. They were then divided into six groups. Two groups were infected with Aspiculuris tetraptera, two groups with Syphacia obvelata, and two groups were left uninfected as controls. At intervals of 2 and 4 weeks after infection, the mice were tested for exploratory activity in a barriered field apparatus. Following the second test, each mouse was necropsied to determine presence of nematodes and to estimate worm burdens. Statistical analysis showed significant depression of exploratory activity in mice harboring S obvelata. No significant depression was found in those harboring A tetraptera.     

Experimental eradication of pinworms (Syphacia obvelata and Aspiculuris tetraptera) from mice colonies using ivermectin.

A spray administration of ivermectin was evaluated for the treatment of pinworm infection in mice. In this study, a spray of 0.1% ivermectin injectable solution over the entire cage once a week, for three consecutive weeks (one cycle treatment), was effective in eradicating both Syphacia obvelata and Aspiculuris tetraptera from mice under experimental conditions. In addition, no acute toxicity was observed in 105 mothers or 687 neonates treated with ivermectin, indicating that ivermectin does not affect murine reproduction. Finally, we attempted to eradicate pinworms from infected mice in our institute using this method. Two cycles of treatment were administered, with a two-week pause between cycles, resulting in complete eradication for at least one year. Treating mouse colonies with spray ivermectin is inexpensive, safe, requires very little labor and is very effective at eradicating pinworms from mice.     

Specific cross-immunity between Hymenolepis nana and H. diminuta: immunization with heterologous and homologous light infections

The consequences of previous and concurrent infection with two related species of cestodes, Hymenolepis nana and H. diminuta, were studied in CD1 mice. A H. diminuta infection strongly affected the establishment and the survival of a secondary H. nana egg or cyst infection administered 30 days later. An infection of 20 H. nana eggs strongly protected against a 5-cyst H. diminuta challenge, whereas an infection of 10 H. nana cysts was ineffective; 20 H. nana eggs also protected against a challenge with 5 cysts of H. diminuta administered 5 days later. No effects were observed in either parasite during a concurrent infection established by administration of cysts. An H. nana egg-infection was unable to affect the establishment of a secondary H. nana cyst-infection given 1 month later; however a significant decrease in growth was found. Similar results were found when a primary H. nana egg-infection was followed 5 days later by the homologous cyst-infection. But an infection with 5 H. nana cysts was unable to protect against a homologous challenge of 5 cysts or 200 eggs. The reciprocal cross immunity between the heterologous parasites and the failure of protection of homologous challenges are discussed in relation to light infections.     

Review of reproductive and developmental toxicity studies with isopropanol

Published studies for reproductive and developmental toxicity conducted with isopropanol have been conducted by the inhalation and oral gavage routes of administration. Interpretation of the data from these studies has resulted in discussions regarding NOAELs and additional benchmark dose modeling publications. Unpublished reproductive and developmental toxicity studies administered in the drinking water were also conducted by BIBRA, and the results of those studies are presented here. In addition, all of the reproductive and developmental toxicity studies conducted with isopropanol are summarized and evaluated for concordance of effects and NOAELs. Endpoints of concern for regulatory agencies were decreases in male mating index and reductions in postnatal pup survival. Original study reports were evaluated and data collated to address these two endpoints, and the data summarized. Data are presented suggesting that there were technical problems in the study that implied a decrease in male mating index, and based on the results from the drinking water studies, the weight of evidence suggests that isopropanol does not affect male mating or fertility at dose levels of up to 1000 mg/kg/day. The weight of evidence suggests that isopropanol can cause decreases in postnatal pup survival following oral gavage administration of 1000-1200 mg/kg/day to the dams. The NOAEL for this endpoint with oral gavage administration was 700 mg/kg/day. Indications of maternal toxicity were also an important predictor for decreased postnatal survival. Decreased postnatal pup survival was also noted in the drinking water studies with isopropanol with a LOAEL of 2278 mg/kg/day and a NOAEL of 1947 mg/kg/day.