Factors affecting the anthelmintic efficacy of papaya latex in vivo: host sex and intensity of infection.

The development of plant-derived cysteine proteinases, such as those in papaya latex, as novel anthelmintics requires that the variables affecting efficacy be fully evaluated. Here, we conducted two experiments, the first to test for any effect of host sex and the second to determine whether the intensity of the worm burden carried by mice would influence efficacy. In both experiments, we used the standard C3H mouse reference strain in which papaya latex supernatant (PLS) consistently shows >80 % reduction in Heligmosomoides bakeri worm burdens, but to broaden the perspective, we also included for comparison mice of other strains that are known to respond more poorly to treatment with papaya latex. Our results confirmed that there is a strong genetic influence affecting efficacy of PLS in removing adult worm burdens. However, there was no effect of host sex on efficacy (C3H and NIH) and no effect of infection intensity (C3H and BALB/c). These results offer optimism that plant-derived cysteine proteinases (CPs), such as these from papaya latex, can function as effective anthelmintics, with neither host sex nor infection intensity presenting further hurdles to impede their development for future medicinal and veterinary usage.

Host genetic influences on the anthelmintic efficacy of papaya-derived cysteine proteinases in mice.

Eight strains of mice, of contrasting genotypes, infected with Heligmosomoides bakeri were studied to determine whether the anthelmintic efficacy of papaya latex varied between inbred mouse strains and therefore whether there is an underlying genetic influence on the effectiveness of removing the intestinal nematode. Infected mice were treated with 330 nmol of crude papaya latex or with 240 nmol of papaya latex supernatant (PLS). Wide variation of response between different mouse strains was detected. Treatment was most effective in C3H (90·5-99·3% reduction in worm counts) and least effective in CD1 and BALB/c strains (36·0 and 40·5%, respectively). Cimetidine treatment did not improve anthelmintic efficacy of PLS in a poor drug responder mouse strain. Trypsin activity, pH and PLS activity did not differ significantly along the length of the gastro-intestinal (GI) tract between poor (BALB/c) and high (C3H) drug responder mouse strains. Our data indicate that there is a genetic component explaining between-mouse variation in the efficacy of a standard dose of PLS in removing worms, and therefore warrant some caution in developing this therapy for wider scale use in the livestock industry, and even in human medicine.

Heligmosomoides neopolygyrus Asakawa & Ohbayashi, 1986, a cryptic Asian nematode infecting the striped field mouse Apodemus agrarius in Central Europe.

BACKGROUND: Heligmosomoides polygyrus is a widespread gastro-intestinal nematode infecting wild Apodemus (wood mice) throughout Europe. Using molecular and morphological evidence, we review the status of Heligmosomoides from Apodemus agrarius in Poland previously considered to be an outlying clade of H. polygyrus, to further resolve the status of the laboratory model species, H. bakeri. METHODS: Morphological analysis of the male bursa and the synlophe, and molecular analyses of concatenated nuclear (28S rDNA, ITS1 and ITS2) and mitochondrial (CO1 and cytb) genes, of Heligmosomoides collected from Apodemus agrarius from two sites in Poland and comparison with related heligmosomids from voles and mice in Eurasia. RESULTS: Heligmosomoides neopolygyrus, a heligmosomid nematode from Apodemus species from China and Japan, is recognised for the first time in western Europe infecting Apodemus agrarius in Poland. It can be distinguished from H. polygyrus by the filiform externo-dorsal rays of the male copulatory bursa and the small, equally distributed longitudinal crêtes on the body. Specimens from A. agrarius are 20% different at ribosomal (ITS1 and ITS2) nuclear loci, and 10% different at the mitochondrial cytb locus from H. polygyrus, and in phylogenetic analyses group with the vole-infecting genus Heligmosomum. CONCLUSIONS: Despite morphological similarity, H. neopolygyrus is only distantly related to H. polygyrus from western European Apodemus, and may be more closely related to vole-infecting taxa. It was brought into Europe by the recent rapid migration of the host mice. Inclusion of H. neopolygyrus in phylogenies makes it clear that Heligmosomoides is paraphyletic, with the pika-infecting Ohbayashinema and the vole-infecting Heligmosomum nesting within it. Clarification of the European status of H. neopolygyrus also allows H. bakeri, the laboratory model species, to be seen as a terminal sister clade to H. polygyrus, rather than as an internal clade of the latter taxon.

Comparing systemic metabolic responses in mice to single or dual infection with Plasmodium berghei and Heligmosomoides bakeri.

Concomitant infections with Plasmodium and gastrointestinal nematodes are frequently observed in humans. At the metabolic level, the cross-talk between the host and multiple coexisting pathogens is poorly characterized. The purpose of this study was to give a comprehensive insight into the systemic metabolic phenotype of mice with a single or dual infection with Plasmodium berghei and Heligmosomoides bakeri. Four groups of eight NMRI female mice were infected with P. berghei or H. bakeri, or with both species concurrently. An additional group remained uninfected, and served as control. Mice were sacrificed at day 19 of the experiment. We collected samples from the liver, spleen, kidney, three intestinal regions, and four brain regions. All biological samples were subjected to (1)H nuclear magnetic resonance spectroscopy, combined with multivariate data analysis, to establish metabolic fingerprints of each tissue from the various infection groups. Compared to uninfected mice, single and dual species infection models showed unique metabolic profiles. P. berghei exerted major effects on glycolysis, tricarboxylic acid cycle, and nucleotide and amino acid metabolism in all studied tissues with the exception of the gut. H. bakeri was characterized by a dysregulation of choline and lipid metabolism in most tissues examined with a particularly strong imprint in the jejunum. Simultaneous co-infection with P. berghei and H. bakeri induced the strongest and most diverse effects in the liver and spleen but led to only minor changes in the intestinal and cerebral parts assessed. Infection with P. berghei showed more pronounced and systemic alterations in the mice metabolic profile than H. bakeri infection. The metabolic fingerprints in the co-infection models were driven by P. berghei infection, whilst the presence of H. bakeri in co-infections had little effect. However, simultaneous co-infection showed indeed the least metabolic disruptions in the peripheral tissues, namely the gut and brain.

Interactive effects of protein nutrition, genetic growth potential and Heligmosomoides bakeri infection pressure on resilience and resistance in mice.

The ability of animals to cope with an increasing parasite load, in terms of resilience and resistance, may be affected by both nutrient supply and demand. Here, we hypothesized that host nutrition and growth potential interact and influence the ability of mice to cope with different parasite doses. Mice selected for high (ROH) or low (ROL) body weight were fed a low (40 g/kg; LP) or high (230 g/kg; HP) protein diet and infected with 0, 50, 100, 150, 200 or 250 L3 infective Heligmosomoides bakeri larvae. ROH-LP mice grew less at doses of 150 L3 and above, whilst growth of ROH-HP and of ROL mice was not affected by infection pressure. Total worm burdens reached a plateau at doses of 150L3, whilst ROH mice excreted fewer worm eggs than ROL mice. Serum antibodies increased with infection dose and ROH mice were found to have higher parasite-specific IgG1 titres than ROL mice. In contrast, ROL had higher total IgE titres than ROH mice, only on HP diets. The interaction between host nutrition and growth potential appears to differentially affect resilience and resistance in mice. However, the results support the view that parasitism penalises performance in animals selected for higher growth.

Genetic growth potential interacts with nutrition on the ability of mice to cope with Heligmosomoides bakeri infection.

Artificial selection for improved productivity may reduce an animal's ability to cope with pathogens. Here, we used Roslin mice, uniquely divergently selected for high (ROH) and low (ROL) body weight, to assess interactive effects of differing growth potential and protein nutrition on host resilience and resistance. In a 2 x 2 x 6 factorial design, ROH and ROL mice were either sham-infected or infected with 250 L(3)Heligmosomoides bakeri and fed diets with 30, 80, 130, 180, 230 and 280 g crude protein per kg. The infected ROL-30 treatment resulted in clinical disease and was discontinued. In the remaining ROL mice, infection and feeding treatments did not affect growth but infection reduced weight gain in ROH-30, ROH-80 and ROH-130 mice. Although infection resulted in temporarily reduced food intake (anorexia) in both mouse lines, mean food intake over the whole experiment was reduced in ROH mice only. ROH mice excreted more worm eggs and had higher worm burdens, with relatively fewer female worms, than ROL mice. However, these resistance traits were not sensitive to dietary protein. These results support the view that selection for high growth may reduce the ability to cope with pathogens, and that improved protein nutrition may to some extent ameliorate this penalty.

FVB/N mice are highly resistant to primary infection with Nippostrongylus brasiliensis.

Nippostrongylus brasiliensis larvae are particularly susceptible to immunological attack during the pre-lung stage of primary and secondary infections in mice. Whilst most of the common laboratory strains of mice are permissive hosts for the parasite, in this study we report for the first time, the strong resistance of naive FVB/N mice to N. brasiliensis. Damage to larvae is evident within the first 24 h of infection and this may be critical to later larval development and reproductive success. Inflammatory responses in the skin, and larval escape from this tissue were comparable in susceptible CBA/Ca and resistant FVB/N mice, with most larvae exiting within 4 h of a primary infection. Lung larval burdens were also similar between strains, but larvae recovered from FVB/N mice were smaller and less motile. In FVB/N mice, larval colonization of the gut was impaired and worms produced very few eggs. However FVB/N mice did not show enhanced resistance to Heligmosomoides bakeri (also known as Heligmosomoides polygyrus), a nematode largely restricted to the gut. Damage done in the pre-lung or lung stages of infection with N. brasiliensis is likely to contribute to ongoing developmental and functional abnormalities, which are profoundly evident in the gut phase of infection.

Helminth communities of an introduced hare (Lepus granatensis) and a native hare (Lepus europaeus) in southern France.

We investigated the parasite communities of introduced Iberian hares (Lepus granatensis) and native European hares (Lepus europaeus) in southern France, where Iberian hares were introduced locally 20 yr ago as a game animal. Parasite communities of sympatric populations of the two hare species and of allopatric populations of European hares were compared. Iberian hares in France harbored a depauperate community of parasites relative to the population in its native habitat in Spain. European hares in areas of sympatry also were infected by Nematodiroides zembrae, which normally infects Iberian hares on their native range.