Investigations on the occurrence of tapeworm infections in German horse populations with comparison of different antibody detection methods based on saliva and serum samples.

BACKGROUND: Effective and sustainable worm control in horses would benefit from detailed information about the current regional occurrence of tapeworms. Different diagnostic methods are currently available to detect Anoplocephala spp. infections in horses. However, the format as well as the sensitivity and specificity of the methods vary considerably. METHODS: A coprological, serological and questionnaire study was conducted to investigate the prevalence and risk factors of tapeworm infections on 48 horse farms in the region of Berlin and Brandenburg, Germany. In total, faecal samples of 484 horses were analysed using the double centrifugation/combined sedimentation-flotation and mini-FLOTAC. Serum (n = 481) and saliva (n = 365) samples were analysed by ELISAs to determine antibody levels against Anoplocephala spp. 12/13 kDa excretory/secretory (E/S) antigens. RESULTS: Cestode eggs were detected in 0.6% of faecal samples (farm prevalence 6.3%) without differences between the two methods. In contrast, antibodies against Anoplocephala spp. were detected in 16.2% (farm prevalence 52.1%) and in 29.5% (farm prevalence 75.7%) of the serum and saliva samples, respectively. Both ELISA based methods for detection of tapeworms reported a greater number of infected animals requiring treatment than were positively identified by coproscopy. Logistic regression analysis identified permanent pasture access, large pastures and regular pasture changes and high strongyle egg counts as risk factors for positive serum antibody responses to Anoplocephala spp. while last treatment with praziquantel was protective. Other protective factors were the presence of foals and high numbers of horses on the farm. Daily removal of faeces from the pasture and horse age did not have a significant effect. CONCLUSIONS: The findings of the present serological investigation indicate that tapeworm prevalence in Berlin/Brandenburg horse farms is much higher than would be anticipated by using conventional/coproscopic analyses. Moreover, the majority of tapeworm-positive horses had not received a cestocidal drug at their last treatment. Considering the already known low sensitivity of the coproscopic detection, the equine veterinary diagnostics can be enhanced by the use of antibody detection methods such as the saliva-based ELISA.

Immune response in spirlins (Alburnoides bipunctatus, Bloch 1782) infested by Ligula intestinalis parasite.

Ligula intestinalis parasite is a cestode that can cause remarkable damages to fishes. SDS-PAGE is one of the methods that can be used to determine the immune serum band polymorphism and immune responses in fishes infested by Ligula intestinalis. This study reports the results of an investigation conducted using SDS-PAGE focusing on immune serum band polymorphism and on the reaction of the immune system in spirlins (Alburnoides bipunctatus) infested by pleurocercoids of Ligula intestinalis parasite. Serum samples from infested spirlins revealed a polymorphism band which differed from that reported in sera of roaches (Rutilus rutilus), a species of the same Cyprinidae family.

[The nature of changes of some immunophysiological characteristics in bream (Abramis brama) infected with plerocercoids (Ligula intestinalis) at various stages of parasite development].

The data from studies of the antimicrobial properties of blood serum, the content of total lipids, and antioxidant activity of immunocompetent tissues and organs of breams Abramis brama infected with plerocercoids Ligula intestinalis depending on the phase of development of the parasite are presented. The quantitative characteristics of the studied parameters are determined.

Equine cestodosis: a sero-epidemiological study of Anoplocephala perfoliata infection in Ethiopia.

A 12/13 kDa antigen, tapeworm ELISA test, developed for use in horses, was used to detect parasite-specific serum antibody, IgG(T), in the serum of donkeys. In a pilot study the 12/13 kDa antigen was tested and proved to detect the antibody, IgG(T), in donkey sera. Blood samples from 797 donkeys, naturally exposed to cestode infection, from four geographical localities were collected and sera were prepared and analysed. There was substantial serological evidence that donkeys were potentially infected with A. perfoliata. A range of ELISA OD values were obtained from the serological assay. Over 26% and 7.5% of the donkeys were moderately and highly infected, respectively, showing at least a 34% sero-prevalence. The rest, 66.1%, were either with low infection intensity or negative for A. perfoliata infection. The risk of infections, both in sero-prevalence and intensity, as determined by ELISA optical density (OD), were highest in the highland areas of Ethiopia where pastures are low-lying and wet, and permanent pasture management is regularly practised. Sex, age and body condition of the donkeys had no significant effect either on prevalence of the infection or on the serum antibody level. These results indicate a risk of intestinal disorders, particularly, colic, associated with A. perfoliata infection in donkeys.

Variation in the reproductive potential of Schistocephalus infected male sticklebacks is associated with 11-ketotestosterone titre.

Parasites can impact host reproduction by interfering with host endocrine systems, but the adaptive nature of such effects is disputed. Schistocephalus solidus plerocercoids are parasites of three-spined sticklebacks Gasterosteus aculeatus that are often associated with impaired host reproduction. Here, we relate reproductive behavior and physiology to levels of the androgen 11-ketotestosterone (11KT) in naturally infected and non-infected male sticklebacks from two UK populations. In one population infected males harbored heavy infections and showed uniformly reduced 11KT titres and kidney spiggin (nesting glue protein) content compared to non-infected fish. However in a second population infection levels were more variable and males with smaller infections recorded 11KT and spiggin titres that overlapped those of non-infected fish; among infected males from this population 11KT and kidney spiggin also both correlated negatively with infection severity. Male reproductive behavior correlated closely with 11KT titre in both populations, and infected males with high 11KT levels exhibited normal reproductive behavior. Our results suggest that Schistocephalus infection per se does not block reproductive development in male sticklebacks, and that some male fish may have the ability to breed whilst infected. Our results are not consistent with the hypothesis that Schistocephalus adaptively castrates male hosts via endocrine disruption; rather they support the hypothesis that reproductive disruption is a side effect of the energetic costs of infection.

[Dynamics of physiological parameters in the nestling of black-backed gull Larus marinus experimentally infested by the cestode Microsomacanthus ductilus (Cestoda: Hymenolepididae)].

The effect of the invasion with the cestode Microsomacanthus ductilus on physiological and biochemical processes in black-backed gull Larus marinus was examined. Experimental invasion of the gull nestling by the cestodes has been performed. Dynamics of the protein, lipid, and carbohydrate metabolism in the time history of the invasion was observed, in comparison with noninfested nestling. Increasing of the content of alpha-globulins and decreasing of the content of protein and albumin in the blood plasma of experimentally infested birds were registered to 4th day after invasion. To 7th day after invasion the level of general lipids and phospholipids decreases, while the content of gamma-globulins and modified form of albumin increases. To 10th day after invasion symptoms of intoxication were observed, but some parameters proved to be reverted to normal condition. So, it can be assumed, that the most intensive reorganization of the metabolism in infested birds takes place in the period between 4th and 7th days after infestation. Possible causes of the observed phenomena are discussed.

Serological changes observed in horses infected with Anoplocephala perfoliata after treatment with praziquantel and natural reinfection.

The serological changes in two groups of horses known to be harbouring Anoplocephala perfoliata were studied; 12 were treated with 1.5 mg/kg praziquantel and 200 microg/kg ivermectin, and 14 were treated with 200 microg/kg ivermectin. Serological and faecal analyses were carried out on each animal at intervals for 758 days. The titres of antibodies specific for A perfoliata decreased from the day of treatment to day 28 in both groups, and continued to decrease in the group treated with praziquantel and ivermectin, with the first significant decrease from the other group at day 121. From day 151 to day 295 the first significant increase in antibody levels in the group treated with both drugs was observed; no A perfoliata eggs were detected in the faeces of these animals until day 295 when five of the 10 were positive.

Influence of intestinal cestodes on the blood picture of the brown rats (Rattus norvegicus) of Hyderabad Sindh, Pakistan.

Present study was performed to determined the changes in the blood picture of the brown rats Rattus norvegicus naturally infected with intestinal cestodes. The results are based on the examination of 82 brown rats captured at random from Hyderabad city and its adjacent areas. Blood profiling of cestode (Hymenolepis diminuta and Hymenolepis nana) infected male and female rats showed significantly lower values of Total Erythrocyte Count (TEC), Haemoglobin (Hb) concentration, Haematocrit (Hct), Mean Corpuscular Haemoglobin (MCH) and Mean Corpuscular Haemoglobin Concentration (MCHC) as compared to their control (non-infected) rats of both sexes. The Mean Corpuscular Volume (MCV) value for infected male and female rats was found significantly higher than those of their control rats. A significantly higher Total Leukocyte Count (TLC) in male and female infected rats was noted as against the non-infected (control) rats of both sexes. The Differential Leukocyte Count (DLC) revealed significant increase in absolute number of each type of cell in cestode infected male and female rats. However, lymphocyte and basophil counts recorded from male infected rats were similar to the counts of their control rats. Conspicuous changes in the blood picture of cestode infected male and female rats indicate host-parasite interaction as well as the triggering of defense mechanism in the host against infection.

Serum malondialdehyde level in children infected with Vampirolepis nana.

BACKGROUND: Vampirolepis nana is the only human tapeworm in which the intermediate host is not necessary and transmission is from person to person. In this study the changes of serum malondialdehyde level, that is, the oxidative stress hypothesis in patients infected with V. nana, was investigated. METHODS: Serum malondialdehyde concentration activity was measured in 32 patients who were positive for intestinal parasite of V. nana. Levels were obtained for the positively infected patients and their age and gender were matched to 32 healthy controls. RESULTS: The difference between malondialdehyde levels of patients infected with V. nana and the control group was statistically significant both for females (P < 0.05) and males (P < 0.05). In the patient and control groups, no correlation was found between age and malondialdehyde levels both in females and males. In addition, no significant correlation could be found between malondialdehyde levels of both females and males for the patient and control groups. CONCLUSION: Malondialdehyde levels were clearly increased in the patients infected with V. nana.

Ligula intestinalis infection as a potential source of bias in the bioindication of endocrine disruption in the European chub Leuciscus cephalus.

European chub Leuciscus cephalus collected from five localities in the lowland and subalpine regions of Austria were analysed for oestrogenic effects of endocrine-disrupting chemicals and the presence of the plerocercoid of the tapeworm Ligula intestinalis. Of 1494 chub analysed, only seven (six males, one female) were found to be infected with single, but large plerocercoids up to 15 cm in length. Ligula-infected fish showed comparatively immature gonads, as demonstrated by the gonadosomatic index and gamete developmental stages. Plasma levels of the egg precursor protein vitellogenin also showed concentrations ranging below the detection limit. The present results indicate that chub infected with L. intestinalis and exposed to exogenous oestrogenic compounds can result in reduced gonadal maturation and produce false oestrogen-positive diagnoses in male fish. For plasma vitellogenin levels, L. intestinalis infections can result in false oestrogen-negative diagnoses in male and female fish.

Identification of a novel eosinophil chemotactic cytokine (ECF-L) as a chitinase family protein.

A novel eosinophil chemotactic cytokine (ECF-L) was purified from the culture supernatant of splenocytes of mice by a combination of anion-exchange chromatography, Procion red-agarose affinity chromatography, size exclusion high performance liquid chromatography (HPLC), and reverse phase HPLC. The NH(2)-terminal amino acid sequence was determined by direct protein sequencing. An ECF-L cDNA clone of 1,506 nucleotides was isolated from a cDNA library, and the nucleotide sequence predicted a mature protein of 397 amino acids. A recombinant ECF-L showed a level of eosinophil chemotactic activity comparable with that of natural ECF-L, and the activity was inhibited by a monoclonal antibody to ECF-L. ECF-L also attracted T lymphocytes and bone marrow polymorphonuclear leukocytes in vitro, whereas it caused selective extravasation of eosinophils in vivo. ECF-L mRNA was highly expressed in spleen, bone marrow, lung, and heart. A comprehensive GenBank data base search revealed that ECF-L is a chitinase family protein. ECF-L retains those amino acids highly conserved among chitinase family proteins, but Asp and Glu residues essential for the proton donation in hydrolysis were replaced by Asn and Gln, respectively. Although ECF-L contains a consensus CXC sequence near the NH(2) terminus akin to chemokine family proteins, the rest of ECF-L shows poor homology with chemokines.

Induction of cerebrospinal fluid eosinophilia in rats by the intraventricular injection of Angiostrongylus cantonensis antigen.

Resistance to Angiostrongylus cantonensis is contingent upon the generation of an eosinophilic response in the CSF of infected hosts. We have studied the parameters required for the generation of this CSF eosinophilia in normally permissive rats. We initially induced a marked peripheral eosinophilia in rats by infection with either Mesocestoides corti or Angiostrongylus cantonensis or the surgical transfer of A. cantonensis young adult worms (YA) into their pulmonary arteries. Next, we injected various antigens into the ventricles of these rats. A. cantonensis-preinfected rats demonstrated significant CSF eosinophilia following injection of A. cantonensis egg antigen, 1st-stage larval (L1) antigen, or M. corti antigen, but not following YA antigen inoculation. A. cantonensis egg and M. corti antigens were potent chemoattractants for eosinophils in an in vitro chemotaxis assay. These data indicate that peripheral eosinophilia, meningeal stimulation by A. cantonensis infection and the presence of potent chemoattractants, e.g., egg and L1 antigens are prerequisites for CSF eosinophil accumulation in permissive rat hosts.

Hepatotrophic activity in mouse serum infected with plerocercoids of Spirometra erinacei.

To investigate the mechanism by which liver weight increases during plerocercoid infections as well as the possible existence of a hepatocyte-growth-factor (HGF)-like substance in the serum of mice infected with Spirometra erinacei plerocercoids, liver DNA synthesis was measured in vivo and in vitro. Infection with S. erinacei plerocercoids significantly stimulated DNA synthesis in mouse parenchymal hepatocytes prior to the increase in liver weight, at least partly by stimulating the induction of the salvage pathways of pyrimidine biosynthesis. Furthermore, infected mouse serum directly stimulated DNA synthesis in cultured mouse parenchymal hepatocytes. These results suggest that an HGF-like substance is present in the serum of mice infected with S. erinacei plerocercoids.

Infection of IL5 transgenic mice with Mesocestoides corti induces very high levels of IL5 but depressed production of eosinophils.

Transgenic mice expressing interleukin 5 (IL5) have been demonstrated to show a lifelong high level eosinophilia. These mice were produced using a construct in which the dominant control region (DCR) of the human CD2 gene was ligated to a 10-kb fragment containing the mouse IL5 gene. The construct allows the expression of the IL5 gene under the control of its own promoter, but the DCR ensures constitutive expression by all T cells. Infection of these transgenic mice with Mesocestoides corti, which is itself a potent inducer of eosinophilia, increases serum IL5 to very high levels. This demonstrates that the transgenes retain inducibility, which is a feature of the endogenous gene. However, despite the high levels of IL5, the numbers of eosinophils in the blood, marrow, and spleen decrease during the period 1-4 weeks after infection. Furthermore, there is a decrease in eosinophil precursors, as assessed by the capacity of bone marrow to produce eosinophils in culture. After this decrease eosinophils return to their previous high levels, although the levels of IL5 remain high. These results suggest that a control mechanism is operating to limit the numbers of eosinophils produced. This control appears to act at the level of the precursor production and may not be directly related to the high levels of IL5.

Eosinophilia in transgenic mice expressing interleukin 5.

Experiments in vitro suggest that although interleukin 5 (IL-5) stimulates the late stages of eosinophil differentiation, other cytokines are required for the generation of eosinophil progenitor cells. In this study transgenic mice constitutively expressing the IL-5 gene were established using a genomic fragment of the IL-5 gene coupled to the dominant control region from the gene encoding human CD2. Four independent eosinophilic transgenic lines have thus far been established, two of which with 8 and 49 transgene copies, are described in detail. These mice appeared macroscopically normal apart from splenomegaly. Eosinophils were at least 65- and 265-fold higher in blood from transgenics, relative to normal littermates, and approximately two- or sevenfold more numerous relative to blood from mice infected with the helminth Mesocestoides corti. Much more modest increases in blood neutrophil, lymphocyte, and monocyte numbers were noted in transgenics, relative to normal littermates (less than threefold). Thus IL-5 in vivo is relatively specific for the eosinophil lineage. Large numbers of eosinophils were present in spleen, bone marrow, and peritoneal exudate, and were highest in the line with the greatest transgene copy number. Eosinophilia was also noted in histological sections of transgenic lungs, Peyer's patches, mesenteric lymph nodes, and gut lamina propria but not in other tissues examined. IL-5 was detected in the sera of transgenics at levels comparable to those seen in sera from parasite-infected animals. IL-3 and granulocyte/macrophage colony-stimulating factor (GM-CSF) were not found. IL-5 mRNA was detected in transgenic thymus, Peyer's patches, and superficial lymph nodes, but not in heart, liver, brain, or skeletal muscle or in any tissues from nontransgenics. Bone marrow from transgenic mice was rich in IL-5-dependent eosinophil precursors. These data indicate that induction of the IL-5 gene is sufficient for production of eosinophilia, and that IL-5 can induce the full pathway of eosinophil differentiation. IL-5 may therefore not be restricted in action to the later stages of eosinophil differentiation, as suggested by earlier in vitro studies.

Parasites of prairie rattlesnakes (Crotalus viridis viridis) and gopher snakes (Pituophis melanoleucus sayi) from the eastern high plains of New Mexico.

Three prairie rattlesnakes (Crotalus viridis viridis) and two gopher snakes (Pituophis melanoleucus sayi) from the eastern high plains of New Mexico (USA) were examined for parasites. One cestode (Oochoristica osheroffi), and two nematode (Kalicephalus inermis and Physoloptera retusa) species were recovered from two infected rattlesnakes. One female gopher snake was infected with two nematode (K. inermis and Rhabdias spp.) and one mite (Entonyssus halli) species.

Effect of Raillietina echinobothrida infection on blood values and intestinal tissues of domestic fowls of Bangladesh.

An examination of the blood profile of domestic fowls infected with Raillietina echinobothrida revealed anaemia with a significant increase of total leukocyte counts and decrease of total serum protein. The pathological changes in the intestines were characterised by villous atrophy, enteritis with cellular infiltration and formation of characteristic granulomas.

Analysis of six serum components from rats infected with tetrathyridia of Mesocestoides corti.

The serum from rats infected intraperitoneally (i.p.) and subcutaneously (s.c.) with the tetrathyridia of Mesocestoides corti was analysed for alpha-amylase, alkaline phosphatase, gamma-glutamyltransferase, glucose, corticosterone and total proteins. A histochemical analysis for hepatic alkaline phosphatase was also carried out. A significant decrease in activity of serum alkaline phosphatase was detected in both the i.p. and s.c. infections, whereas a large increase in hepatic alkaline phosphatase was observed in i.p. infections. A significant increase in serum corticosterone was found in both the i.p. and s.c. infections. No changes were detected in the other four components. The relevance of these findings is discussed with respect to the host-parasite relationship.

Pathophysiology of Mesocestoides corti infection in the mouse.

Liver histology and serum enzyme and protein changes were studies in two strains of mice showing different initial susceptibilities to infection with Mesocestoides corti. The results show an increase in ALT and AST levels during the period of invasion and proliferation in the liver and a decrease in the levels of these two enzymes following encapsulation of the parasite in the liver and liver regeneration. A progressive loss of albumin was accompanied by increases in the levels of the beta- and gamma-globulins. These changes are discussed in the light of our knowledge of the effects of this parasite upon its host.