Developmental surface ultrastructure of Macroorchis spinulosus in albino rats

Developmental surface ultrastructure of Macroorchis spinulosus was studied by scanning electron microscopy. One-day-old juvenile fluke was leaf-shaped and bent ventrally. Body surface was covered densely with peg-like spines and with cobblestone-like cytoplasmic processes. Ciliated sensory papillae were concentrated around oral sucker. Several unciliated sensory papillae occurred equidistantly on oral sucker and acetabulum. The ciliated papillae appeared in two longitudinal lines symmetric bilaterally on dorsal surface. On adult flukes, tegumantal spines became wider in middle of the body surface. The cytoplasmic processes differentiated into more fine velvety form. It is likely that the differentiated fine cytoplasmic processes are an increased absorptive surface to adult M. spinulosus. It is suggested that single pointed tegumental spines on anterior half of the body may be supportive for this fluke to migration.     

Morphology of inhibited larvae of the bovine lungworm Dictyocaulus viviparus.

Some morphological features of inhibited fourth stage (L4) and fourth moult larvae (4M) of the bovine lungworm Dictyocaulus viviparus are described. Inhibition was induced by maintaining the third stage larvae (L3) for 6 weeks at 4 degrees C. Inhibited fourth stage (L4) and fourth moult larvae (4M) were collected by perfusion of the lungs of experimentally infected calves after necropsy at 15 and 68 days post infection (d.p.i.), respectively. Inhibited 4M, isolated at 68 d.p.i., were about ten times larger than inhibited L4 isolated at 15 d.p.i.     

Mucosal mast cell responses in the small intestine of rats infected with Echinostoma hortense

Mucosal mast cell (MMC) responses and worm recovery rates in rats infected with Echinostoma hortense were investigated from day 3 to day 56 post-infection (p.i.). Experimental infected group showed apparently higher number of MMC in each part of the small intestine than that of the control group. The number of MMC in the duodenum increased gradually after the infection and reached a peak on day 35 p.i. Thereafter, the number of MMC continued to decrease at a slow pace. The kinetics of MMC responses in the upper and lower jejunum were similar to that of the duodenum, but the number of MMC in the jejunum was lower. The worm recovery rate decreased with respect to time of which it was markedly reduced on day 49 and 56 p.i. The duration in which a high number of MMC appeared was similar to that in which a low rate in worm recovery was recorded. These results indicate that intestinal mastocytosis may play an important role in the expulsion of E. hortense.     

Mucosal mast cells in Sprague-Dawley rats infected with Hymenolepis diminuta tapeworms

Six-week-old Sprague-Dawley female rats each infected with 40 Hymenolepis diminuta cysts showed increased mastocytosis from day 30 post-infection (p.i.) to day 47 p.i. Rats treated on day 40 p.i. with anthelmintic and autopsied 22 days later showed reduced mucosal mast cell (MMC) counts. Other infected rats, treated with anthelmintic on day 40, challenged with a 10 cysticercoid infection on day 47 and subsequently autopsied between day 8 and 19 post-challenge, maintained a high MMC count. Age of rats in this experiment was not a factor in mastocytosis.     

The effect of ethanol upon early development in mice and rats. XIV. The effect of acute intoxication with beer and wine upon preimplantation development in rats

The preimplantation effect of acute intoxication during the preimplantation period of pregnancy (i.p. or i.v. injection on day 4) with beer and wine has been investigated on female albino rats (Wistar strain). The following criteria were applied for checking preimplantation development: mean number of embryos/animal; topographical distribution of the embryos; developmental stage attained; occurrence of pathological embryonic forms. The control on day 5 of pregnancy revealed no significant effect upon the developmental criteria used. The data obtained are compared with our own previous results.     

Experimental infection of Apis mellifera honeybees with Nosema ceranae (Microsporidia)

In this report, an experimental infection of Apis mellifera by Nosema ceranae, a newly reported microsporidian in this host is described. Nosema free honeybees were inoculated with 125,000 N. ceranae spores, isolated from heavily infected bees. The parasite species was identified by amplification and sequencing the SSUrRNA gene of the administered spores. Three replicate cages of 20 honeybees each were prepared, along with one control cage (n=20) supplied with sugar syrup only. The infection rate was 100% at the dosage administered. The presence of Nosema inside ventricular cells was confirmed in the samples using ultrathin sectioning and transmission electron microscopy. By day 3 p.i. a few cells (4.4%+/-1.2) were observed to be parasitized, whereas by 6 days p.i. more than half of the counted cells (66.4%+/-6) showed different parasite stages, this value increasing on day 7 p.i. (81.5%+/-14.8). Only one control bee died on day 7 p.i. In the infected groups, mortality was not observed until day 6 p.i. (66.7%+/-5.6). Total mortality on day 7 p.i. was 94.1% in the three infected replicates and by day 8 p.i. no infected bee was alive. After the infection, the parasites invaded both the tip of folds and the basal cells of the epithelium and the autoinfective capacity of the spores seemed to spread the infection rapidly between epithelial cells. On day 3 p.i., mature spores could be seen inside host cell tissue implying that the developmental cycle had been completed. The large number of parasitized cells, even the regenerative ones, the presence of autoinfective spores and the high mortality rate demonstrate that N. ceranae is highly pathogenic to Apis mellifera. Possible relation with bee depopulation syndrome is discussed by authors.     

Immunity to Litomosoides carinii in Mastomys natalensis. II. Effects of chemotherapeutically abbreviated and postpatent primary infections on challenges with various stages of the parasite

Naive Mastomys natalensis, Litomosoides carinii-infected M. natalensis at a postpatent stage of the infection and L. carinii-infected M. natalensis treated chemotherapeutically with furazolidone (FUR), FUR and diethylcarbamazine (FUR/DEC) or amoscanate (AMOS) were challenged by either injection or implantation of 40 third stage larvae (L3, s.c.), 40 fourth stage larvae (L4, 16 days old, i.p.), 20 male and 20 female preadult worms (36 days old, i.p.), 12 adult female worms (i.p.) or 6 X 10(6) microfilariae/kg (i.v.). Microfilaraemia in animals challenged at a postpatent stage (independent of the kind of challenge), was either totally suppressed or at least greatly reduced. Necropsy of L3-challenged animals showed that neither the length of the worms nor their content of morphologically intact, intrauterine stages was affected. Infected, treated animals challenged with developing stages (L3, L4 and preadult worms) showed reduced levels of microfilaraemia (by up to 75%). Dissection of AMOS-treated, L3-challenged animals showed that both the developmental rate and the fertility of the worms were affected. Microfilaraemia was also reduced after implantation of adult worms into treated animals. This was independent of the interval between treatment and challenge (44-150 days) except in animals challenged 10 days after AMOS-treatment, which showed no difference from naive controls. However, infected, treated M. natalensis, cotton rats and gerbils did not develop immunity against intravenously injected blood microfilariae.     

Therapeutic potential of the methanolic extract of Lepidium sativum seeds on mice infected with Trypanosoma evansi

The present study aimed to investigate the therapeutic potential of the methanolic extract of Lepidium sativum seeds in mice experimentally infected with Trypanosoma evansi. A total of thirty-two male Swiss albino mice were randomly divided into four groups: the first group was the normal control, while the second, third and fourth groups were infected intraperitoneally with 1 × 10⁴ trypanosomes. The third and fourth groups were treated with 100 μl of Lepidium sativum seed extract (LSSE) at a dose of 200 mg/kg body weight intraperitoneally (infected + LSSEI) and orally (infected + LSSEO) respectively, once a day, for a period of four days.Parasitaemia was found to be significantly raised in the untreated infected group, reaching 2 × 10⁷ at day 4 post-infection, but was significantly reduced by 65.5% and 88% in the mice treated orally and intraperitoneally with LSSE, respectively. The erythrocyte count, HCT, haemoglobin content, leucocyte count and the percentage of lymphocytes was significantly reduced in the untreated infected group, while the treatment with LSSE returned these parameters to their pre-infection values. In addition, our study proved that LSSE provided protection against liver tissue damage and decreased the levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT). The present study also established that intraperitoneal injection of LSSE is more effective than oral administration in the treatment of trypanosome infection in mice. In conclusion, the infection caused haematological, biochemical and histological changes that were ameliorated following treatment with LSSE.     

Hexavalent chromium effects on motor activity and some metabolic aspects of Wistar albino rats

The effects of hexavalent chromium on the motor activity and some metabolic aspects of albino rats were studied. 0.07 g Cr(VI)/l administered in drinking water produced no effects on the motor activity of rats, at least over 28 days of treatment. 0.7 g Cr(VI)/l in drinking water and 2 mg Cr(VI)/kg body wt injected i.p. significantly decreased the motor activity of rats after 7 days and 1 day, respectively. The treated rats showed a metabolic disadvantage in relation to the controls. The effects caused by Cr(VI) on the motor activity in rats could be an indication of the neurotoxicity of this metal.     

The increase in mannose receptor recycling favors arginase induction and Trypanosoma cruzi survival in macrophages

The macrophage mannose receptor (MR) is a pattern recognition receptor of the innate immune system that binds to microbial structures bearing mannose, fucose and N-acetylglucosamine on their surface. Trypanosoma cruzi antigen cruzipain (Cz) is found in the different developmental forms of the parasite. This glycoprotein has a highly mannosylated C-terminal domain that participates in the host-antigen contact. Our group previously demonstrated that Cz-macrophage (Mo) interaction could modulate the immune response against T. cruzi through the induction of a preferential metabolic pathway. In this work, we have studied in Mo the role of MR in arginase induction and in T. cruzi survival using different MR ligands. We have showed that pre-incubation of T. cruzi infected cells with mannose-Bovine Serum Albumin (Man-BSA, MR specific ligand) biased nitric oxide (NO)/urea balance towards urea production and increased intracellular amastigotes growth. The study of intracellular signals showed that pre-incubation with Man-BSA in T. cruzi J774 infected cells induced down-regulation of JNK and p44/p42 phosphorylation and increased of p38 MAPK phosphorylation. These results are coincident with previous data showing that Cz also modifies the MAPK phosphorylation profile induced by the parasite. In addition, we have showed by confocal microscopy that Cz and Man-BSA enhance MR recycling. Furthermore, we studied MR behavior during T. cruzi infection in vivo. MR was up-regulated in F4/80+ cells from T. cruzi infected mice at 13 and 15 days post infection. Besides, we investigated the effect of MR blocking antibody in T. cruzi infected peritoneal Mo. Arginase activity and parasite growth were decreased in infected cells pre-incubated with anti-MR antibody as compared with infected cells treated with control antibody. Therefore, we postulate that during T. cruzi infection, Cz may contact with MR, increasing MR recycling which leads to arginase activity up-regulation and intracellular parasite growth.     

Trypanosoma lewisi: comparative activity of a feral isolate in two strains of rats assessed by measurement of cell population, reproductive development and respiratory activity.

Lee C. M., Harris L. M. and Aboko-Cole G. F. 1978. Trypanosoma lewisi: comparative activity of a feral isolate in two strains of rats assessed by measurement of cell population, reproductive development and respiratory activity. International Journal for Parasitclogy8: 187–192. Comparative respiratory behaviour, population growth and host antibody formation were examined for a feral isolate of Trypanosoma lewisi grown in albino and black rats. Oxygen uptake was measured for endogenous, glucose or normal homologous rat serum substrates. Trypanosomes from albino rats respired 72% faster endogenously, 70% faster with glucose, and 60% faster with rat serum than parasites from black rats. Greater oxygen consumption of T. lewisi from albino rat hosts correlated well with the fact that the trypanosome developed greater parasitemias in albino than in black rats. Parasitemias were patent longer in albino rats and peak populations were found 2 days later in these hosts than in black rats. On the average, higher coefficients of variability in cell length continued five days longer in albino rats than in black animals. Delay in reproductive development was indicated by continued low variability in trypanosome cell sizes in albino and black rats.     

Life Cycle and Mating Behavior of Belonolaimus longicaudatus in Gnotobiotic Culture

The life cycle of Belonolaimus longicaudatus was observed in vitro on excised roots of Zea mays. Roots were cultured on Gamborg''s B5 medium in petri dishes with 1.5% agar adjusted to pH 5.8 and incubated at 28 °C in darkness. Second-stage juveniles (J2) fed on the roots and started the second molt (M2) to the third-stage juveniles 2 days after inoculation (DAI). The third molt (M3) to the fourth-stage juveniles occurred 7 DAI, followed by the fourth molt (M4) to males 13 DAI or to females 14 DAI. Nematode gender differences were observed by the end of the fourth molt. The first male appeared 15 DAI and the first female 17 DAI, after which mating occurred. Males were attracted to females, and mating was observed. Mating was required for reproduction. Fertilized females began to lay eggs 19 DAI and continued egg laying without the further presence of males during a 90-day observation. All of the eggs hatched. Unfertilized females rarely laid eggs, and none of the eggs were able to hatch. Feeding took place between each molt and before egg deposition occurred. The first-stage juveniles molted in the eggs 4 days after deposition, and J2 hatched from eggs 5 days after egg deposition. The life cycle from J2 to J2 was completed in 24 days.     

EFFECTS OF MICROWAVES AND ELF MAGNETIC FIELD ON THE PHAGOCYTIC ACTIVITY OF VARIOUSLY TREATED RAT MACROPHAGES

The purpose of this study was to investigate the effects of 9450-MHz microwaves and extremely low frequency magnetic fields (ELFMF) on the phagocytic activity of rat macrophages in control rats and those treated with vitamins C and E. In the microwave group, 24 albino Wistar rats were exposed to microwaves (2.65 mW/cm², specific absorption rate [SAR]: 1.80 W/kg) for 1 h/day for 21 days. Thirty-two albino Wistar rats were divided into four groups (one control, three experimental) (n = 8). The rats in the first exposure group were only exposed to microwaves for 1 h per day for 21 days. In addition to exposure with microwaves as in the first experimental group, vitamins E and C (150 mg/kg/day) were injected intraperitoneally into the rats in the second and third exposure groups, respectively. In the magnetic field exposure group, 26 albino Wistar rats were divided into two groups: the sham (n = 12) and exposed groups (n = 14). The rats in the experimental group were exposed to ELFMF (50 Hz, 0.75 mT) for 3 h/day for 3 weeks. After completing the exposure period, the rats were sacrificed under ketalar anesthesia. The viability of isolated alveolar macrophages of rats in the microwave and ELF groups was determined and compared to sham groups. The results were analyzed with the Mann–Whitney U test. In the microwave group, the phagocytic activity in the experimental groups was found to be higher than the sham groups. However, with phagocytic activity in rats treated with both microwaves and vitamins, only the vitamin C group was significant (p < 0.05). In the magnetic field group, the phagocytic activity of rats exposed to ELFMF was lower than that of the sham group, but the results were not significant (p > 0.05). Rectal temperatures of microwaveexposed groups were found to be significantly higher compared to the control group (p < 0.05).     

Early behavioral changes and quantitative analysis of neuropathological features in murine prion disease: Stereological analysis in the albino Swiss mice model

Behavioral and neuropathological changes have been widely investigated in murine prion disease but stereological based unbiased estimates of key neuropathological features have not been carried out. After injections of ME7 infected (ME7) or normal brain homogenates (NBH) into dorsal CA1 of albino Swiss mice and C57BL6, we assessed behavioral changes on hippocampal-dependent tasks. We also estimated by optical fractionator at 15 and 18 weeks post-injections (w.p.i.) the total number of neurons, reactive astrocytes, activated microglia and perineuronal nets (PN) in the polymorphic layer of dentate gyrus (PolDG), CA1 and septum in albino Swiss mice. On average, early behavioral changes in albino Swiss mice start four weeks later than in C57BL6. Cluster and discriminant analysis of behavioral data in albino Swiss mice revealed that four of nine subjects start to change their behavior at 12 w.p.i. and reach terminal stage at 22 w.p.i and the remaining subjects start at 22 w.p.i. and reach terminal stage at 26 w.p.i. Biotinylated dextran-amine BDA-tracer experiments in mossy fiber pathway confirmed axonal degeneration and stereological data showed that early astrocytosis, microgliosis and reduction in the perineuronal nets are independent of a change in the number of neuronal cell bodies. Statistical analysis revealed that the septal region had greater levels of neuroinflammation and extracellular matrix damage than CA1. This stereological and multivariate analysis at early stages of disease in an outbred model of prion disease provided new insights connecting behavioral changes and neuroinflammation and seems to be important to understand the mechanisms of prion disease progression.Key words: prion disease, optical fractionator, neuropathology, behavioral changes, albino Swiss mice     

Periodic appearance of a predominant variant antigen type during a chronic Giardia lamblia infection in a mouse model

Giardia lamblia (syn. Giardia duodenalis, Giardia intestinalis) infections are associated with continuous antigenic variation of the parasite which is mediated by the parasite's major surface antigen, named variant surface protein. Offspring mice and corresponding mothers were infected with G. lamblia clone GS/M-83-H7 (expressing variant surface protein H7) and various parameters of this infection were assessed in a long-term follow-up investigation. Our experimentation revealed that variant surface protein H7-type trophozoites were replaced by new variant-type trophozoites during the early stage of infection (around day 8 p.i.), but the original variant-type re-emerged at at least two time-points during the later stages of infection (at days 22 and 42 p.i.). Such periods of variant surface protein H7-type trophozoite re-expansion were accompanied by transient production of intestinal IgA against variant-specific epitopes on a 314-aa N-terminal region of variant surface protein H7. At late stages of infection (between days 42 and 200 p.i.), most mice produced intestinal IgA against both variant surface protein H7 and other antigens of the parasite. At these stages, infection seemed to be resolved in most mice, but occasional reappearance of relatively high (at day 64 p.i.) or at least detectable (at days 80 and 120 p.i.) amounts of intestinal parasites indicated that G. lamblia GS/M-83-H7 infections in mice may enter into a latent chronic phase which is interrupted by sporadic breakthroughs of parasite growth.     

Extraintestinal Migration of Centrorhynchus sp. (Acanthocephala: Centrorhynchidae) in Experimentally Infected Rats

Reptiles were known to serve as paratenic hosts for Centrorhynchus (Acanthocephala: Centrorhynchidae) in Korea, but the infection course in experimental animals was not elucidated yet. In this study, the tiger keelback snakes (Rhabdophis tigrinus) were collected and digested with artificial pepsin solution, and the larvae of Centrorhynchus were recovered from them. Then, the collected larvae were orally infected to rats for developmental observations. In rats, all the larvae were observed outside the intestine on day 3 post-infection (PI), including the mesentery and abdominal muscles. As for the development in rats, the ovary of Centrorhynchus sp. was observed at day 15 PI, and the cement glands were 3 in number. Based on the morphological characteristics, including the arrangement of proboscis hooks, these larvae proved to be a species of Centrorhynchus, and more studies were needed for species identification.     

Observations on the infectivity and fecundity of Schistosoma curassoni from Senegal in albino mice

An average of 11% of adult Schistosoma curassoni were recovered from 200 albino mice which had been infected subcutaneously with 150-250 cercariae. Worms were primarily found in the portal veins. The average number of intrauterine eggs per female during the first 100 days p.i. was 13 and the average number of eggs produced per female worm was 103 per day for the first 60 days post oviposition. The majority of eggs were recovered from the liver (98.3%). The oograms were determined until day 95 p.i. For screening of antischistosomal drugs it is recommended not to use infections older than 60 d.p.i.     

Immunity to Litomosoides carinii in Mastomys natalensis. I. Effect of immunization with microfilariae and existing primary infections on the parasitaemia after microfilariae injection and challenge infection

Subcutaneous injections of intrauterine stages of Litomosoides carinii into Mastomys natalensis induced strong immunity to i.v. injected blood microfilariae. Immunity, developed after boostering with an i.p. and an i.v. injection of microfilariae, did not totally suppress the parasitaemia of a challenge infection but reduced significantly the microfilaraemia level. No effect was found on number and size of the worms of the challenge infection, the number of microfilariae or the number of leucocytes in the pleural cavity. Delayed type hypersensitivity reactions in challenged animals were similar to those in non-immunized, infected controls. Sera of immunized animals agglutinated microfilariae and mediated cell attachment to microfilariae. Challenge infections did not change this until the end of the fourth week post infection but sera taken 32 days after challenge and later failed to induce such reactions. Challenge infections performed 120 or 240 days after a primary infection did not increase the parasitaemia of recipients. Dissections carried out 130 days after the challenge showed that (a) the developmental rate of the challenge infection was reduced by about 50%; (b) the size of the challenge parasites was reduced; and (c) that these worms produced significantly less embryonic stages in comparison to worms of primary infections, of which about 90% were abnormal.     

Progression of Root-knot Nematode Symptoms and Infection on Resistant and Susceptible Cottons

Progressive development in cotton root morphology of resistant A623 and susceptible M-8 cotton (Gossypium hirsutum L.) lines following infection by the root-knot nematode Meloidogyne incognita was studied in glass front boxes. Symptom development and radicle growth were observed; degree of galling, gall and egg mass diameter, and number of eggs per egg mass were recorded; and root segments were examined histologically. Small cracks caused by M. incognita appeared in the root epidermis and cortex soon after the cotyledons expanded on day 4. The cracks were longer and wider and extended through the cortex when the first true leaf became visible at day 8. Galls had formed on taproots by this time. When exposed to M. incognita, A623 had faster radicle growth (22%), fewer and smaller cracks in the root epidermis and cortex, fewer and smaller root galls, one-twelfth as many egg masses, and one-fourth as many eggs per egg mass as M-8. Root cracking, galling, and giant cell formation are major effects of M. incognita that may predispose cotton roots to pathogens resulting in synergistic interactions and diseases.