An optimum protocol for the cryopreservation of third-stage larvae (L3) of Angiostrongylus cantonensis was determined by: (i) use of different dimethyl sulphoxide (DMSO) concentrations; (ii) use of different FBS concentrations in RPMI-1640 medium; and (iii) variation in the thawing temperature. Studies showed that with Concentrations of DMSO at 4·8% and FBS at 40% and a thawing temperature of 43°C, L3 survival assessed by motility was highest at 56%. The infectivity of the cryopreserved L3 of A. cantonensis was 20% compared to controls obtained from slugs.

The effect of calcium and ammonium chloroacetates in increasing the yield of potatoes grown in Ayrshire fields infected with Heterodera schachtii is demonstrated.

From the examination of roots of treated and untreated plants it is believed that this increase in yield is due to a reduction in the eelworm infection, and from hatching experiments this reduction is thought to be the result of the action of calcium chloroacetate in retarding and reducing the hatching of larvae from the cysts.

Elevated serum lipids are associated with infections of laboratory rodents with plerocercoids of Spirometra mansonoides. The effect of infection with these larval tapeworms on triglyceride degradation and hepatic de novo fatty acid synthesis was investigated in Syrian hamsters. Serum lipoprotein electrophoresis revealed a consistent elevation in very low density lipoproteins in the infected animals. Lipoprotein lipase activity was enhanced in the infected animals. After seven days of plerocercoid infection the activity of acetyl-CoA carboxylase (E.C. 6.4.1.2) was significantly elevated after 6,12 and 18 hours of fasting. Fatty acid synthetase was significantly increased after 0,6,12,18 and 24 hours of fasting. Therefore, a chronic insulin-like activity on lipid metabolism of hamsters is associated with plerocercoid infection.

A low molecular weight fraction (30 KDa) of the cattle filarial parasite Setaria digitata that was earlier demonstrated to have allergenic activity was characterized to be a zinc-dependent cysteine proteinase. Immediate type hypersensitivity (ITH) reaction to the proteinase was evaluated in lymphatic filariasis patients and in endemic controls from Orissa, India. The extent of ITH positivity to the proteinase in infected individuals ranged from 20% in chronic filariasis (CP) patients group to 56% in asymptomatic microfilaraemic carriers (AS). About 62% of endemic normals (EN) were also ITH positive. The serum levels of IgG subclasses were compared in ITH positive and ITH negative filarial patients (AS and CP) as well as in endemic normals (EN) respectively. IgG4 levels were found to be inversely dependent on ITH reaction only in AS groups. Asymptomatic patients (AS) with positive ITH reactivity had lower IgG4 than ITH negative individuals from the same group. The serum levels of other IgG subclasses except IgG2 did not correlate with ITH reactivity. IgG2 levels were higher in ITH negative EN and CP patients but not in the AS group.

On 23/3/43 a small experiment was sown with commercial onion seed at Kirton, being one of a series laid down at county oentres in order to study the effect of different times of lifting on the development of wastage in store. Four varieties, which may be designated A, B, R, U, were sown in strips of three rows 10in. apart, in triplicate, the rows being 41 yards long and forming three randomised blocks. The liftings were also in randomised block pattern, 3 × 3, across the length of the rows. Two sub-plots of each variety were taken for convenience, consisting of adjacent six-foot lengths across the three rows, the produce of each sub-plot being put into a separate box immediately on lifting.

The life-cycle of Tremiorchis ranarum Mehra and Negi, 1926 (Trematoda: Plagiorchiidae) infecting Rana tigrina and R. cyanophlyctis is elucidated. Life-cycle stages, from egg to egg-producing adults, were successfully established in the laboratory. Life-cycle took more than two months for completion. Cercariae were found in the snail, Pila virens collected from paddy fields at Chelembra in the Malappuram district of Kerala during the monsoon months. Cercariae are of the xiphidiocercous type. Metacercariae occurred beneath the skin and in head, tail and body musculature of tadpoles of R. tigrina and R. cyanophlyctis. Adult frogs are refractory to cercarial infection. Frogs became infected when they fed on tadpoles containing metacercariae. Successive stages in the development to adult from the metacercariae were followed. Chronological synchronization in the reproductive activity of the host and the parasite is briefly discussed.

Four new genera (each containing one new species) belonging to the order Rhabditida have been described from Jodhpur Sewer. Mesodiplogasteroides n.g. (subfamily Diplogasterinae) is distinguished from the closely allied genus Butlerius by the absence of setose papillae on the head and the presence of a rudimentary bursa in male. Operculorhabditis n.g. (subfamily Rhabditinae) is characterized by the presence of a flap-like, deciduous, cuticular operculum covering the vulva which is well posterior. Praeputirhabditis n.g. (subfamily Rhabditinae) is unique in possessing prepuce-like loose cuticle through which the tail can be protruded. Saprorhabditis n.g. (sub-family Protorhabditinae) has neither a glottoid apparatus nor a median oesophageal bulb but possesses an oesophageal collar. The genus Matthesonema is transferred to the subfamily Rhabditinae. Rhabditis (Rhabditella) octopleura is being recorded for the first time from India.

The anterior end of the miracidium of Fasciola hepatica contains a large flask-shaped apical gland and four unicellular lateral glands, all of which have ducts which pass to the tip of the apical papilla. These glands appear to be involved in penetration of the larva into the snail host. The apical gland secretes as the miracidium proves the epidermis of the host before attachment. It seems likely that its secretion is a chemical which lyses the epidermal cells. The lateral glands are PAS-positive and may contain a neutral mucopolysaccharide. They also secrete as the miracidium probes the snail and a layer of PAS-positive material may be seen at the leading edge of the apical papilla as the larva penetrates into the host. Both the apical gland and the lateral glands may be visible in the sporocyst for several days after penetration.

Metacercariae identified as those of the heterophyid trematode, Galactosomum ussuriense were collected from 3 species of brackish water fishes from Bheemunipatnam. The organization of the the larva is described in detail. This is the first record of the metacercaria of this species although the adult has not so far been recorded from India.

A method for hatching second-stage larvae of Toxocara canis is described. Whole adult worms are processed by centrifugation in the presence of hypochlorite which allows the grinding step to be eliminated. The technique is more rapid and a high yield is obtained.

In 1928 Taylor put forward the theory that “…Syngamus trachea is a species within which there may be found a number of strains showing a peculiar physiological adaptation to the host in which they occur.” He had not had much success in a number of experiments in which he had tried to infect chickens with S. trachea from starlings. The first experimental feedings resulted in only light infections though control experiments with the chicken “strain” resulted in much heavier ones. After the first passage through the chicken however, the gapeworms had become more adapted to the new host, and could produce much more serious infections in the next generation.

Extracts prepared from the microbivorous nematode Panagrellus redivivus and the plant-parasitic nematode Meloidogyne incognita were used to provide general protease activities for peptide substrate screening and species comparisons. Each extract was evaluated for its ability to degrade a broad range of nematode FMRFamide-like peptides (FLPs), key regulatory messengers governing nematode growth and development. Clear quantitative differences between the two extracts were observed using FMRFamide as a substrate. Extract potency assessed at EC50 (μg/μ l extract protein for 50% substrate digestion) was 1.8-fold greater for P. redivivus than for M. incognita, and potency assessed at EC90 was 2.5-fold greater. An overall potency difference was also present when screening the digestion of 17 nematode FLPs, but it was not universal. The mean percentage digestion of eight of the 17 FLPs was greater (P < 0.02) with P. redivivus extract (76.3 ± 8.2) than with M. incognita extract (38.1 ± 8.7), but the means for the other nine FLPs were not different. Three FLPs (KPSFVRFa, AQTFVRFa, RNKFEFIRFa) were degraded extensively by the extracts of both species, and two FLPs (SAPYDPNFLRFa, SAEPFGTMRFa) were degraded 2.9-fold and 5.3-fold greater, respectively, with M. incognita extract than with P. redivivus extract. The ability of each extract to degrade FMRFa and KSAYMRFa was significantly reduced by using peptide analogues containing single d-amino acid substitutions, and the substitution effects were positional. Both FMRFa and KSAYMRFa were competitive substrates for aminopeptidases in each extract, but only the competitive ability of FMRFa was reduced by d-amino acid substitution. The variety and complexity of nematode FLP degradation by preparations representing phylogenetically and developmentally different nematode sources are discussed.

Carbon-13 nuclear magnetic resonance (NMR) spectroscopy was employed to investigate alterations in hepatic carbohydrate metabolism in Meriones unguiculatus infected with Echinococcus multilocularis. Following portal vein injections of an equimolar mixture of ]#x005B;1,2-13C2]acetate and [3-13C]lactate, perchloric acid extracts of the livers were prepared and NMR spectra obtained. Isotopomer analysis using glutamate resonances in these spectra showed that the relative contributions of endogenous and exogenous substrates to the acetyl-CoA entering the tricarboxylic acid cycle differed significantly between infected and control groups. The mole fraction of acetyl-CoA that was derived from endogenous, unlabelled sources (FU) was 0.50±0.10 in controls compared to 0.34±0.04 in infected animals. However, the fraction of acetyl-CoA derived from [3-13C]lactate (FLL) was larger in livers of infected animals than those from controls with values of 0.27±0.04 and 0.18±0.04, respectively. Similarly, the fraction of acetyl-CoA derived from [1,2-13C2]acetate (FLA) was larger in livers of infected animals compared to those in controls; the fractions were 0.38±0.01 and 0.32±0.07, respectively. The ratio of FLA:FLL was significantly smaller in the infected group with a value of 1.42±0.18 compared to 1.74±0.09 for the controls. These results indicate that alveolar hydatid disease has a pronounced effect on the partitioning of substrates within the pathways of carbohydrate metabolism in the host liver.

Several taxonomic studies of the entomopathogenic nematode family Steinernematidae have reported on the minute tail projections on the tail tips of females of the first generation; however, the occurrence and morphology of these traits in other steinernematid species and their taxonomic significance are still unclear. In the present study, the tail projections in females of seven steinernematid species belonging to four phylogenetic clades were examined. The projections are present in all of the tested species and their number and shape is consistent within species but significantly differs among species. It thus seems that these structures may have taxonomic significance at the species level. Based on these results, we suggest that every description of a new species in the family Steinernematidae should contain a precise morphology of the female tail, focused on these projections, using both light and scanning electron microscopy.

A literature survey was undertaken in order to draw up a definitive list of helminth parasites of the wolf, Canis lupus. From 27 papers a total of 72 helminth species from 40 genera were recorded that infect wolves, of which 93% were identified from the gastrointestinal tract at necropsy. They comprised 28 species of nematode, 27 species of cestode, 16 species of trematode and one acanthocephalan. Of these, 46 species were able to be included in further meta-analysis of prevalence data derived from 25 publications for which the total number of wolves examined was 1282 (1066 from Nearctic populations, and 216 from the Palaearctic region). These two populations were further subdivided into three relevent ecosystems or biomes, i.e. temperate/montane (n=216), boreal (n=805) or tundra (n=261). The meta-analysis of relative prevalence indicated the most common helminth species to be the tapeworm Taenia hydatigena, which occurred at relative rates of >30% for either zoogeographic region as well as in each of the three biomes. The related tapeworm, Echinococcus granulosus also exhibited high meta-prevalence (>19%) in all host biomes. The hookworm Uncinaria stenocephala was the most prevalent nematode species by meta-analysis (meta-prevalence 44.9%) in the temperate/montane biome, while the ascarid Toxascaris leonina was the dominant helminth species (meta-prevalence 73.9%) in the tundra wolf populations. Trematodes in the genus Alaria were the dominant fluke (meta-prevalence 3–5%) in all biomes. Analysis of published studies for helminth biodiversity using the Shannon-Wiener index based on species number and meta-prevalence by region or biome, indicated that highest helminth diversity occurred in wolf populations of the temperate/ montane biome (Palaearctic), and was lowest in tundra wolf populations of the Nearctic (P<0.05). Helminth species assemblage in European wolf populations was therefore at least as great or more varied than was recorded for the larger less disturbed wolf populations of North America.

The histopathological effects of Taenia crassiceps infection or T. crassiceps metacestode factor inoculation on the mouse ovary were determined using six female mice in three groups: infected mice, mice inoculated with the metacestode factor and control mice. The control group was subcutaneously inoculated with healthy peritoneal fluid. The infected group was intraperitoneally inoculated with 40 T. crassiceps metacestodes, and the metacestode factor group was subcutaneously inoculated with T. crassiceps metacestode factor (MF). Light and electron microscopy and TUNEL (terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labelling) assays revealed a significant increase in ovarian follicular atresia (predominantly in antral/preovulatory stages of development), oocyte degeneration (P< 0.05), and a decrease in the amount of corpus luteum in follicles of mice infected and inoculated with MF compared with the control group. Significant abnormalities of the granulosa cells and oocytes of the primordial, primary and secondary ovarian follicles occurred in both treated mouse groups (P< 0.05) compared with no degeneration in the control group. These pathological changes in female mice either infected with T. crassiceps metacestodes or inoculated with T. crassiceps MF may have consequences for ovulation and fertility.

Until now, four species of the Trichinella genus have been identified in Europe: Trichinella spiralis, T. nativa, T. britovi and T. pseudospiralis. The aim of this work was to establish a sound polymerase chain reaction (PCR)-based method to differentiate these four species using mitochondrial rDNA as a reliable genetic marker and to evaluate the sensitivity of this method. Full-length DNA sequences coding for the small and large mitochondrial rRNA (mt-rrnS and mt-rrnL) of the four species are described. A multiplex PCR was designed and successfully tested on 24 European isolates. As few as one larva, or 100 pg of genomic DNA was detected, providing equivalent sensitivity to previously described PCR methods. The PCR-based method of mitochondrial rDNA amplification was thereby established as a sensitive and reproductive diagnostic method for the four European Trichinella species.

Two new species of Syphacia (Nematoda: Oxyuridae) are described from endemic murids of Sulawesi, Indonesia: Syphacia(Syphacia)taeromyos sp. n. and S.(S.)paruromyos sp. n. parasitic in the caecum of Taeromys celebensis and Paruromys dominator, respectively. They are readily distinguished from all of the congeners recorded from Indonesia–Australian regions by having a round cephalic plate, vesicular lateral alae in the male, posteriorly positioned excretory pore in the male, and/or lacking cervical alae. Syphacia (S.) paruromyos differs from S. (S.) taeromyos by having a whip-like tail appendage in the male, longer relative distance between excretory pore and vulva, and larger eggs. The round cephalic plate in both sexes and developed vesicular lateral alae in the male are morphological traits common in endemic Syphacia species hitherto known from Sulawesi murids, suggesting that they have derived from a common ancestor and evolved with their hosts in the isolated insular environment.

Seasonal, host sex and age-related variations in helminth egg and coccidian oocyst counts were investigated in a naturally infected wild bushbuck (Tragelaphus scriptus) population in Queen Elizabeth National Park, western Uganda from April 2000 to February 2002. The prevalence and mean intensity quantified as the number of eggs and oocysts per gram of faeces were taken as a measure of parasite burdens. Host sex and age-related differences in prevalence values were not found but the overall prevalence of Eimeria sp. was significantly higher during the rainy season, and peak counts were recorded either during or soon after a peak rainfall. A similar trend was observed for Moniezia spp., although the results were marginally not significant. There were also no significant differences in mean intensity values, relative to host sex, age or season.