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Impact of protein energy malnutrition on Trichuris suis infection in pigs concomitantly infected with Ascaris suum.

The objective of this experiment was to investigate a possible interaction between protein and energy malnutrition (PEM) and intestinal nematode infections. We report on a 3 x 2 factorial study in which pigs were fed either a low protein energy (LPE), low protein (LP) or a normal protein energy (NPE) diet, and 6 weeks later inoculated with Trichuris suis (4000 infective eggs). Secondarily, in order to obtain a polyparasitic status, pigs were concomitantly inoculated with Ascaris suum (600 infective eggs). The number of T. suis-infected pigs was higher in LP pigs compared with NPE pigs (100 versus 58%; P = 0.037), although the differences in median T. suis worm burdens between groups at necropsy 10 weeks post-infection (p.i.) (LPE: 795; LP: 835; NPE: 48 worms; P = 0.33) were not significant. Interestingly, only T. suis in NPE were highly aggregated (k = 0.44), in contrast to a more uniform distribution among pigs in LPE (k = 1.43) and LP (k = 1.55) i.e. the majority of pigs harboured moderate worm burdens in LPE and LP, while most pigs had few or no worms in NPE. Further, T. suis worms in the LPE and LP groups were decreased in length (mean: LPE: 23.5 mm; LP: 24.3 mm; NPE: 29.4 mm; P = 0.004). The pre-patency period of T. suis was also extended in the LPE and LP groups, as reflected by lower faecal egg output at week 6 (P = 0.048) and/or 7 p.i. (P = 0.007). More A. suum worms were recovered from LP compared with the NPE group (mean: 5.4 versus 0.6; P = 0.040); this was accompanied by a higher faecal egg output in the former (P = 0.004). The low protein diets resulted in lower pig body weight gains, serum albumin, haemoglobin and packed cell volume (PCV) levels as well as diminished peripheral eosinophil counts. Infection significantly altered these parameters in the low protein groups, i.e. the pathophysiological consequences of infection were more severe in the PEM pigs. These results demonstrate that reduced protein in the diet leads to malnourishment of both the host pigs and T. suis, and compromises the pig's ability to resist infection by T. suis and A. suum.

Animals↗

Long-term survey on intestinal nematode and cestode infections in stray puppies Ibaraki Prefecture.

We conducted the survey on intestinal nematode, mainly Toxocara canis, and cestode infections in stray puppies aged less than 5 months in Ibaraki Prefecture for 8 years from 1988 to 1995. The rate of T. canis infection stayed almost unchanged for the 8 years and ranged from 72.2 to 85.4%. Of the total 916 dogs, 732 (79.9%) harboured with T. canis. The average rates of other helminthic infections showed 24.6 (Diphylidium caninum), 2.5 (Trichuris vulpis), 1.2 (Ancylostoma caninum) and 0.7% (Spirometra erinaceieuropaei) respectively.

Animals↗

Previous infection with the nematode Nippostrongylus brasiliensis alters the immune specific response against Chlamydophila abortus infection.

An experimental mouse model to analyze the interaction between the immune responses elicited following infection with Nippostrongylus brasiliensis and Chlamydophila abortus has been established. Mice infected with C. abortus 7 days after N. brasiliensis showed an increased bacterial multiplication in spleen and liver compared to bacteria-alone infected mice. However the morbidity of these mice, expressed as weight loss, was significantly lower. Analysis of the immune responses elicited showed that spleen from co-infected mice had reduced IFN-gamma production in response to C. abortus antigen. The bias towards a type 2 response in co-infected mice was confirmed by an increase in the production of IL-4 and in the lower ratio IgG2a/IgG1. In pregnant mice co-infection caused a delay in the time of abortion and an increased systemic susceptibility to C. abortus infection.

Animals↗

Eicosanoids in parasites and parasitic infections.

Eicosanoids are lipid mediators with multiple functions in vertebrate tissues and invertebrate organisms. In this review the roles of eicosanoids--mostly prostaglandins (PGs), thromboxanes and leukotrienes--in parasite physiology and host-parasite interactions are discussed. PGs are present in the saliva of blood-sucking arthropods facilitating feeding by increasing local blood flow and prolonged attachment of ticks by immune suppression. Release of various eicosanoids has also been demonstrated for a number of protozoan and metazoan endoparasites. These substances appear to play a role in penetration, immune suppression, inflammation or modulation of haemostasis, enabling parasite invasion and establishment. Moreover, endogenous eicosanoids serve various functions in parasite metabolism and physiology. In many parasitic infections eicosanoids are involved in host pathology, e.g. granuloma formation, coagulopathy, secretory diarrhoea, or fever. Immune suppression by induction of PG release, in particular PGE2, by host defence cells appears to be a common feature of many parasitic infections and is though to be important for parasite establishment. Contradictory results have been obtained for gastrointestinal nematode infections, which probably reflect the considerable differences between the various models employed. Although most of the available studies indicate an important role for eicosanoids in parasites and parasitic infections, our current knowledge is still fragmentary and more data are urgently needed.

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Genetic analysis of mortality in murine angiostrongyliasis costaricensis using SMXA recombinant inbred mouse strains.

The SMXA recombinant inbred mouse strain set was produced by systematic inbreeding from the F2 generation of a cross between two progenitor inbred strains, A/J and SM/J, which differed markedly with respect to the patterns of infection with Angiostrongylus costaricensis. We have applied this set to genetic analysis of mouse susceptibility to this nematode infection. The mortality was variable among substrains of the SMXA RI strains, indicating the involvement of multiple genes. Linkage analysis showed several chromosomal regions closely linked to mortality; chromosome 6 (D6Rik86, 87; P<==0.001), 10 (D10Rik66-D10Mit12; P=0.0058), 13 (D13Rik79, 80; P=0.0096) and 17 (D17Mit28-D17Rik76; P=0.0088).

Angiostrongylus↗

Antigenic peptides on the surface of Dirofilaria immitis microfilariae.

Various methods of radioiodination were employed to identify peptides on the surface of Dirofilaria immitis microfilariae. Optimum surface radiolabelling occurred with the lactoperoxidase-catalyzed reaction. Two major peptides of 16 and 14 kDa were labelled by this method. These peptides were soluble in Nonidet P-40, were not glycosylated, and showed no signs of disulfide linkages. These peptides were immunoprecipitated by sera from D. immitis-infected dogs, but not by sera from uninfected dogs or sera from dogs with potentially cross-reactive nematode infections. Analysis of the 14 and 16 kDa peptides by two-dimensional gel electrophoresis revealed that the 16 kDa peptide was a single unit with a pI of 5.25 whereas the 14 kDa band was composed of three individual peptides with pI values ranging from 5.6 to 6.1. Iodination by chloramine T resulted in the same panel of labelled peptides but suffered from poor efficiency of 125I incorporation. The viability of microfilariae labelled by the standard Bolton-Hunter method decreased by 50% following the reaction which resulted in the labelling of a variety of internal components.

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Nematode resistance in plants: the battle underground.

Parasitic nematodes infect thousands of plant species, but some plants harbor specific resistance genes that defend against these pests. Several nematode resistance genes have been cloned in plants, and most resemble other plant resistance genes. Nematode resistance is generally characterized by host plant cell death near or at the feeding site of the endoparasitic worm. The timing and localization of the resistance response varies with the particular resistance gene and nematode interaction. Although there is genetic evidence that single genes in the nematode can determine whether a plant mounts a resistance response, cognate nematode effectors corresponding to a plant resistance gene have not been identified. However, recent progress in genetics and genomics of both plants and nematodes, and developments in RNA silencing strategies are improving our understanding of the molecular players in this complex interaction. In this article, we review the nature and mechanisms of plant-nematode interactions with respect to resistance in plants.

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Activation of jird (Meriones unguiculatus) macrophages by the filarial parasite Brugia pahangi.

Peritoneal macrophages from Mongolian jirds (Meriones unguiculatus) with either lymphatic or intraperitoneal infections of Brugia pahangi were studied to determine the effects of infection on macrophage function and morphology. Macrophages were collected at 40, 90, 140, and 200 days after inoculation of infective third-stage larvae and assayed for phagocytic and bactericidal activity by the acridine orange method and for morphological changes by light and electron microscopy. Significant increases in phagocytic and microbicidal activity (P less than or equal to 0.01) were observed in peritoneal macrophages collected from jirds with intraperitoneal infections when compared with peritoneal macrophages from jirds with lymphatic infections and resident peritoneal macrophages from normal, noninfected jirds. Morphological changes in peritoneal macrophages from jirds with intraperitoneal infections were similar to those found in thioglycolate-elicited macrophage populations. Granuloma formation was also observed in the peritoneal cavities of intraperitoneally infected jirds. The peritoneal cavity may serve as a model to study cell-worm interactions in filarial nematode infections.

Animals↗

The combined effect of fenbendazole treatment and a move to aftermath 7 or 9 weeks after turnout on Dictyocaulus viviparus infections in calves.

A grazing study was performed with the main objective of examining the effect of fenbendazole (FBZ) in a 'dose and move' system on nematode infections in calves with special emphasis on Dictyocaulus viviparus. Three groups of six calves were grazed from May to October 1993. One group (DM7) was treated with FBZ and moved to aftermath (pasture which had only been mown) 7 weeks after turnout. The second group (DM9) was similarly treated and moved 9 weeks after turnout and the third group served as untreated pasture control group (PC) and was moved to aftermath 9 weeks after turnout. FBZ treatment removed adult lungworms from DM7 and DM9. Tracer calves grazed during the first 7 or the first 9 weeks after turnout acquired mean burdens of 18 and 125 lungworms, respectively. In PC faecal larval counts increased until the end of August. Most of the animals in this group were then suffering from lungworm disease and emergency treatment with ivermectin was given. In both FBZ-treated groups, larvae reappeared in the faeces of some of the calves 4-5 weeks after treatment. Subsequent reinfection resulted in higher mean faecal larval counts in both groups 2 months after treatment, although variation in faecal larval counts was high. In DM7 values tended to be higher than in DM9. These higher larval counts were associated with mild signs of parasitic bronchitis in some calves of DM7, whereas no signs were seen in DM9. At the end of the experiment, all calves, and also a group of six permanently housed non-infected control calves (HC), were infected experimentally with 5000 D. viviparus larvae to evaluate development of immunity. The worm counts at necropsy showed that all calves on pasture had developed immunity.

Animal Feed↗

In vitro activity of antimicrobial agents against the endosymbiont Wolbachia pipientis.

Arthropod-transmitted (filarial) nematodes are important causes of disease in humans in tropical countries, yet no safe drug appropriate for mass delivery kills the adult worms. However, most filarial nematodes contain rickettsia-like bacteria of the genus Wolbachia, and related bacteria also occur in insects. There is increasing evidence that these bacteria have significant functions in the biology of filarial nematodes. They are thus important targets in the search for antifilarial drugs and experiments in animals and humans have suggested that antibiotic therapy has potential in treating filarial infections. To optimize future clinical trials there is a need for a fast and simple in vitro drug screen to compare drug efficacies against Wolbachia. In the absence of Wolbachia-infected nematode cell lines, we have utilized an Aedes albopictus insect cell line, naturally infected with Wolbachia, to test the activity of antimicrobial agents. Of the five antibiotics tested, doxycycline, oxytetracycline and rifampicin showed good activity (MICs of 0.0625, 4 and 0.0625 mg/L, respectively) whereas ciprofloxacin and penicillin were shown to have no effect.

Animals↗

Infectivity of entomopathogenic nematodes (Steinernematidae and Heterorhabditidae) to female ticks of Boophilus annulatus (Arachnida: Ixodidae).

Exposing Boophilus annulatus (Say) to different concentrations of Steinernema carpocapsae (Filipjev) infective juveniles in petri dishes (50-10,000 nematodes per dish) resulted in high mortality (greater than 90%) at nematode concentrations as low as 500 nematodes per dish within 8 d. At a concentration of 10,000 nematodes per dish, 100% of the ticks died within 2 d after infestation. After exposure to 500 nematodes per dish, complete mortality was achieved with the Heterorhabditis bacteriophora strain 'HP88' within 4 d. During the same period, only 15 and 40% mortality were recorded with the 'Mexican' and 'All' strains of S. carpocapsae, respectively. In a lethal dose analysis, S. carpocapsae strain 'DT' was the most infective strain with the lowest LD50 and LD90 values (15 and 165 infective juveniles per tick, respectively). The 'All' strain of S. carpocapsae was the least infective of the four strains tested, with LD50 and LD90 values of 372 and 9,251 infective juveniles/tick, respectively. Optimal temperature for tick control by the nematodes was between 22 and 26 degrees C. Mortality rate was reduced at 18 and 30 degrees C. The susceptibility of fully engorged ticks was not influenced by the weight of the replete females. Nematode infection did not have an adverse effect on egg laying by surviving ticks.

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The soybean cyst nematode, Heterodera glycines: a genetic model system for the study of plant-parasitic nematodes.

Despite advances in understanding plant responses to nematode infection, little information exists regarding parasitic mechanisms. Recently, it has become possible to perform genetic analysis of soybean cyst nematode. Integration of classic and reverse genetics and genomic approaches for the parasite, with host genetics and genomics will expand our knowledge of nematode parasitism.

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Parasitic infections of the intestine.

Intestinal protozoal infections cause significant disease not only in the tropics but also in immunocompromised hosts and returning travellers in the developed world. Precise diagnosis of protozoal intestinal infection by microscopy can be difficult. Enzyme immunoassays for antigen detection are being used for some protozoal infections with some limitations which will, it is hoped, be overcome by molecular techniques. Nucleic acid amplification techniques could help improve detection of microsporidial species, which are difficult to detect microscopically and allow differentiation between Entamoeba histolytica and Entamoeba dispar. Definitive treatment for Cryptosporidium parvum infection remains elusive but new drugs are being evaluated. Cessation of thiabendazole production should not affect therapy of intestinal nematode infections as potent alternative therapy is available.

Animals↗

Identification of antigens with potential for immunodiagnosis of Parelaphostrongylus tenuis and Elaphostrongylus cervi infections in red deer (Cervus elaphus elaphus).

Red deer (Cervus elaphus elaphus) were infected experimentally with Parelaphostrongylus tenuis in New Brunswick, Canada, and with Elaphostrongylus cervi in New Zealand. Excretory-secretory (E-S) antigens from adult P. tenuis were evaluated for their serodiagnostic potential in identifying P. tenuis and heterologous E. cervi infections in a Western blot. The antigen recognition profile of sera from animals infected with P. tenuis varied between individuals and with duration of infections, whereas that of pooled sera from animals infected with E. cervi showed less variation. A single molecule of 42-43 kDa was recognized consistently by sera from all animals infected with either P. tenuis or E. cervi. Sera from unexposed control deer and from those with other heterologous nematode infections did not consistently identify this antigen. Serorecognition of the 42-43-kDa antigen by deer infected with P. tenuis resulted in a sensitivity of 99% and a specificity of 85% (> or =1 mo postinfection). Although antibody to this antigen waned with time, the persistence of recognition up to 34 mo postinfection with P. tenuis exemplifies its diagnostic value. The sensitivity and specificity of diagnosis using this molecule were each 100% for identifying deer infected with E. cervi (> or =3 mo postinfection). Two other molecules from E-S of adult P. tenuis, 26-28 and 10-12 kDa, were also diagnostic, although their recognition was not persistent throughout infections. These 2 molecules may prove useful in combination with the 42-43-kDa antigen to help identify all infected animals during all phases of infections. This research represents the first conclusive identification of antigens with real potential for reliable antemortem immunodiagnosis of both P. tenuis infections and heterologous E. cervi infections.

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Specialization of the entomopathogenic nematode Steinernema scapterisci with its mutualistic Xenorhabdus symbiont.

The level of specialization of the entomopathogenic nematode Steinernema scapterisci with its native Xenorhabdus symbiont was investigated by testing (1) the influence of non-native bacterial strains on nematode fitness within an insect-host (Galleria mellonella) and (2) specificity of the association between the nematode infective juveniles and non-native bacteria. All non-native Xenorhabdus spp. or Photorhabdus spp. strains tested were mutualistically associated with other entomopathogenic nematodes in nature. We showed that most of the Xenorhabdus spp. strains tested led to an insignificant difference of the nematode's fitness compared to the one obtained with the native bacterium. Conversely, Photorhabdus spp. strains almost entirely abolished nematode reproduction. The phylogenetic analysis of bacterial strains tested, showed that there was a negative correlation between S. scapterisci's reproduction rate with a bacterial strain and the genetic distance of this bacterial strain from the native one. We also showed that the native bacterium was the only one which was transmitted by S. scapterisci's infective juveniles. All these results, suggested a specialization between S. scapterisci and its native Xenorhabdus. As the same phenomenon was already demonstrated in the association between S. carpocapsae and X. nematophila, specialization between partners would not be an exception in entomopathogenic nematode-bacteria interactions. Nevertheless, S. scapterisci showed a dramatically higher compatibility with non-native Xenorhabdus spp. strains than did S. carpocapsae, suggesting differences in the co-evolutionary processes between nematodes and bacteria in these two model systems.

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Naturally occurring variability in some phenotypic markers and correlates of haemonchotolerance in West African Dwarf goats in a subhumid zone of Nigeria.

West African Dwarf (WAD) goats of the Nigerian subhumid zone generally show strong resistance and resilience to Haemonchus contortus in laboratory experiments, although a relatively small proportion are susceptible to infection. Little is known about these extremes of response phenotype in nature. Therefore, a survey was carried out of gastrointestinal nematode infections in WAD goats, with emphasis on abomasal worms, at three goat markets in Southern Nigeria during the rainy season. Faecal samples (n=1070) were collected weekly from goats between April and September, and 352 abomasa and small intestines from local abattoirs were examined. Total strongyle (prevalence=65.0%) and H. contortus (prevalence=64.3%) faecal egg counts (FEC) varied between the three markets, being highest throughout at Opi. FEC increased from April to peak in August. Based on raw FEC, 76.1% of goats had FEC of <100, and 4.7%>500. Adjustment of these figures for monthly and between-market differences, gave figures of 78.8 and 3.4%, respectively. H. contortus worm burdens (WB) showed a similar pattern with 67.9% of goats harbouring <200 worms and 8.2% >1000, and after adjustment 69.6 and 6.0%, respectively. Fecundity, based on eggs in the uterus, did not vary between markets or monthly, but fell with increasing WB. Trichostrongylus colubriformis was less frequent (prevalence=42.4%) but goats from Opi also carried higher WB, and worms were similarly highly aggregated in hosts. When the between-market and monthly differences for both species were controlled, a highly significant positive correlation between the species emerged. Therefore, although a small subset of goats, highly susceptible to H. contortus, exists in this breed, the majority show resistance under field conditions and the resistant phenotype is also resistant to T. colubriformis. Both species are highly aggregated in the susceptible subset of the population. While, we cannot yet exclude alternative explanations, our data are compatible with a strong genetic basis for this phenomenon.

Abomasum↗

Effects of rhizobacteria and root symbionts on the reproduction of Meloidogyne javanica and growth of chickpea.

The effects of rhizobacteria, i.e. Pseudomonas fluorescens, Azotobacter chyroococcum and Azospirillum brasilense, alone and in combination with root symbionts, Rhizobium sp. and Glomus mosseae, on the growth of chickpea, Cicer arietinum, and reproduction of Meloidogyne jaranica were studied. When added alone G. mosseae was better at improving plant growth and reducing galling and nematode reproduction than any other tested organism. Application of P. fluorescens caused an almost similar increase in plant growth to that caused by Rhizobium sp., while use of A. chroococcum was better than A. brasilense in improving growth of nematode --infected plants. Combined use of P. fluorescens with G. mosseae was better at improving plant growth and reducing galling and nematode multiplication than any other combined treatment.

Animals↗

N6-methyladenine DNA modification modulates pathogen virulence in nematodes.

Understanding the global regulatory mechanisms that control pathogen virulence gene expression is essential for elucidating the molecular basis of pathogenicity. N6-methyladenine (6 mA) plays a crucial role in regulating gene expression in response to various environmental stresses; however, its role in pathogen virulence remains largely unexplored. Here, we report the widespread occurrence of 6 mA across 17 nematode isolates and map its genomic landscape in six notorious agriculturally important pathogen root-knot nematodes (RKNs). We demonstrated that 6 mA is characterized by a conserved GAG motif across nematodes, but exhibits species-specific distribution patterns and distinct effects on gene expression. In particular, its enrichment in transposable elements (TEs) differs between polyploid and diploid nematodes, suggesting lineage-specific epigenetic regulation potentially associated with polyploidy. We further identified two functional 6 mA demethylases, MiNMAD-1 and MiNMAD-2, and confirmed their catalytic activity and active sites. Host-induced gene silencing (HIGS) of minmad-1 significantly increased plant resistance to three polyploid RKN species. A detailed functional analysis revealed that minmad-1 knockdown broadly affected gene expression during the parasitic stage, including genes involved in virulence, thereby reducing nematode infectivity. Together, our findings suggest 6 mA demethylase as a key epigenetic regulator of RKNs' virulence, providing new insights into nematode biology and offering promising targets for the development of sustainable control strategies.

Animals↗