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Biomedical subjects

P Nansen

Publications and source records attributed to P Nansen.

At least 109 records · Page 6Linked to original sources

The influence of high- and low-fibre diets on the activity of piperazine against Oesophagostomum spp. in pigs.

An in vivo study evaluated the effect of diet on the efficacy of piperazine against nodular worms of pigs. Twenty pigs, later allocated into five groups, were each infected (and 37 days later re-infected) with 3000 infective larvae of a mixed isolate of Oesophagostomum dentatum and Oesophagostomum quadrispinulatum. Beginning on day 23 post infection (p.i.), pigs in groups 1 and 2 were fed a low-fibre diet consisting of 70% barley flour and 30% protein concentrate, while pigs in groups 3, 4 and 5 were fed a high-fibre diet consisting of 55% barley flour, 21% oat-husk meal and 24% protein concentrate. On day 42 p.i., pigs in groups 1 and 3 were orally dosed with 200 mg piperazine dihydrochloride (Ascarex D, 53%) per kg bodyweight, the recommended dose, while pigs in group 4 were given 100 mg kg-1. Groups 2 and 5 served as non-treated controls for the respective dietary regimens. Eight days after treatment, the pigs were slaughtered and worms recovered from the caecum and large intestine (divided into five sections) and counted. The mean worm count reduction (WCR) in group 1 (full-dose piperazine with low-fibre diet) was 89.8%, while the high-fibre diet in group 3 increased the WCR to 99.4%. In group 4, where the pigs were fed the high-fibre diet and treated with only 100 mg piperazine kg-1, the WCR was 90.9%, identical to the "low fibre" group 1 treated with twice this piperazine dose. There was a zero efficacy recorded against immature worms in all three treated groups. The high-fibre diet improved the efficacy of piperazine against more pathogenic and generally more tolerant O. quadrispinulatum to 99.2% compared with 84.3% at the low-fibre diet.

Animals↗

Biological control of gastro-intestinal nematodes--facts, future, or fiction?

The potential of using fungi to prevent nematodosis caused by parasites with free-living larval stages is well documented today. In this respect Duddingtonia flagrans, a net-trapping, nematode-destroying fungus, appears to be the most promising candidate. Laboratory experiments and in-vivo studies, where fungal spores have survived passage through the gastro-intestinal tract of cattle and horses, plus field studies with cattle, horses and pigs, demonstrate significant reduction in the number of infective larvae that develop in the faecal environment. In field trials this reduction subsequently leads to reduced infectivity of herbage and also reduced worm burdens in grazing animals. A status of the present situation, primarily based upon work performed in Denmark within the last 6-8 years, plus an outlook for practical implementation of an integrated control strategy including the use of nematode-destroying fungi in the future is discussed.

Animals↗

A cross-sectional study of helminths in rural scavenging poultry in Tanzania in relation to season and climate.

A cross-sectional study on the prevalence of helminths in rural scavenging poultry on six hundred adult chickens selected randomly from six villages in the Morogoro Region, Tanzania during the wet and the dry seasons, was conducted. The trachea and gastrointestinal tract of each bird were examined for the presence of parasites. All chickens were infected with one or several species of helminths, the average being 4.8 +/- 1.7 helminths per chicken during the wet season and 5.1 +/- 1.8 during the dry season. A total of 29 different helminth species were shown in the trachea or the gastrointestinal tract. The following species were identified: Syngamus trachea (0.7% (wet season), 2% (dry season)); Gongylonema ingluvicola (6.3%, 17.7%); Tetrameres americana (54.3%, 60.3%); Dispharynx nasuta (0%, 2.7%); Acuaria hamulosa (8.3%, 19.3%); Ascaridia galli (28.3%, 32.3%); Heterakis gallinarum (74.0%, 78.7%); H. isolonche (18.7%, 5.0%); H. dispar (25.7%, 6.3%); Allodapa suctoria (40.0%, 52.0%); Subulura strongylina (3.3%, 1.0%); Strongyloides avium (0.3%, 3.0%); Capillaria annulata (2.0%, 0.0%); C. contorta (9.0%, 1.0%); C. caudinflata (2.0%, 4.3%); C. obsignata (8.7%, 25.0%); C. anatis (4.0%, 9.0%); C. bursata (1.0%, 2.7%); Raillietina echinobothrida (41.3%, 46.3%); R. tetragona (25.3%, 21. 3%); R. cesticillus (8.7%, 2.7%); Choanotaenia infundibulum (0.0%, 3. 7%); Hymenolepis carioca (9.0%, 18.0%); H. cantaniana (48.0%, 43. 0%); Amoebotaenia cuneata (39.3%, 36.0%); Metroliasthes lucida (1.0%, 0.3%); Davainea proglottina (5.7%, 0.3%) and Polymorphus boschadis (0.3%, 0.0%). No trematodes were found. No correlation was found between season and prevalence, or season and mean worm burdens. Twelve helminths of the species recovered represent new local records.

Animals↗

The effect of concurrent or sequential Oesophagostomum dentatum and O. quadrispinulatum infections on the worm burdens of the two species in pigs.

The interaction between the 2 nodular worm species in the pig, Oesophagostomum dentatum (O.d.) and Oesophagostomum quadrispinulatum (O.q.), was studied by comparing the development and distribution of the species following single or mixed infections. The faecal egg excretion levels were assessed at regular intervals from week 3 post-inoculation, and indicated a strong negative impact of the introduction of O.q. on the continued egg excretion of O.d. All pigs were killed 9 weeks after the first inoculations to determine the composition and location of the worm burdens in the large intestine. O.q. was found more anteriorly located in the intestine than O.d., thus confirming previous descriptions. When both species were present, the distribution of O.d. was moved further posteriorly and was more spread out than in single-species infections. There appeared to be no adverse effect of O.d. on the establishment and fecundity of O.q. However, the worm recoveries corroborated the egg excretion observations, namely reduced worm burdens of O.d. if O.q. was introduced, or if O.q. was already present. It is uncertain whether this effect is caused by differences in host reaction against the two species, or whether a more specific competition occurs between the two nodular worm species in pigs.

Animals↗

Schistosoma japonicum and Trichuris suis infections in pigs fed diets with high and low protein.

The aim of the study was to measure the impact of Schistosoma japonicum and Trichuris suis infections in young growing pigs fed low- or high-protein diets. Thirty-two pigs. 6-10 weeks old, were randomly allocated to 2 groups receiving either a high- or a low-protein diet. After 11 weeks half of the pigs from each group were infected with 1500 S. japonicum cercariae and 4000 T. suis eggs. The weight of the pigs was measured throughout the study, and blood and faecal samples were collected every second week from the time of infection. At the time of infection the low-protein pigs had significantly lower mean body weights, haemoglobin and albumin levels compared with the high-protein pigs, and this pattern continued throughout the study. The serum albumin concentration was further significantly reduced in the infected low-protein pigs compared to the non-infected low-protein pigs. Significantly more S. japonicum worms as well as faecal and tissue eggs were found in the low-protein pigs compared with the high-protein pigs. No differences between the 2 diet groups were observed in T. suis establishment rates or faecal egg excretion. We conclude that this low-protein diet increased the establishment rates of S. japonicum, favoured larger deposits of S. japonicum eggs in the liver and faecal egg excretion, reduced weight gains and caused anaemia and hypoalbuminaemia in young growing pigs as compared with a high-protein diet.

Animals↗

Sex-manipulated Ascaris suum infections in pigs: implications for reproduction.

In spite of the vast distribution and considerable impact on human and animal health of Ascaris suum and A. lumbricoides, little is known of the sexual biology and reproductive capacity of these intestinal nematodes. By oral transfer of adult female worms to previously parasite-naive pigs we show that in the absence of males the egg production ceases after 2-3 weeks. Such females readily resume egg production a few days after oral transfer of male worms. These observations throw light on an important aspect of Ascaris biology, but also pave the way for possible experimental interbreeding between the human and pig Ascaris species.

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Experimental Ascaris suum infection in the pig: worm population kinetics following single inoculations with three doses of infective eggs.

To study population kinetics during primary Ascaris suum infections, 3 groups of 52 pigs each were inoculated with 100, 1000, or 10,000 infective eggs. In all groups, the majority of larvae was found in the liver on day 3 post inoculation (p.i.) and in the lungs on day 7 p.i. Liver white spots, caused by migrating larvae, were most numerous at day 7 p.i., whereafter they gradually healed, and only low numbers of granulation-tissue type white spots and lymphonodular white spots persisted at days 21-56 p.i. Independent of dose level, 47-58% of the inoculated eggs were recovered as larvae in the small intestine on day 10 p.i., but most larvae were eliminated at days 17-21 p.i. This elimination started earlier and removed a higher percentage of the worms with increasing inoculation dose, resulting in small strongly aggregated worm populations by day 28 p.i. (k of the negative binomial distribution was low: 0.2-0.4) without significant differences between groups. Thus, overdispersion, which is a characteristic of both porcine and human ascarosis, is found here under experimental conditions where aggregation factors like host behaviour, transmission rate, host status etc have been partly or totally controlled.

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Experimental Oesophagostomum dentatum infections in the pig: worm populations at regular intervals during trickle infections with three dose levels of larvae.

A trickle infection experiment was undertaken to study in detail the population dynamics of Oesophagostomum dentatum in pigs. Three groups of 32 pigs were inoculated via the feed twice weekly with 100 (Group A), 1000 (Group B) or 10,000 (Group C) O. dentatum infective larvae (L3). Five pigs from each group were killed 2, 4, 8, 12, 16, and 20 weeks after the first inoculation (p.i.) to determine their worm burdens. Weekly faecal egg counts were determined. At slaughter, worms were counted, differentiated according to sex and developmental stage, and their length measured. Faecal egg counts ranked with dose rate until week 15, but later were more variable. The proportion of the total number of L3 administered which were recovered at slaughter inversely ranked with dose rate. In group C it decreased over time, whereas in groups A and B there was no consistent pattern. Worm fecundities (epg/female) in groups A and B were higher than in group C. The lengths of the female worms increased over time, whereas the lengths of the male worms remained approximately constant from week 8. The study suggests reduced establishment of incoming larvae and lower fecundity of the female worms at high dose levels.

Animals↗

Comparison of two different methods for inactivation of viruses in serum.

In order to compare protocols for inactivation of viruses potentially present in biological specimens, three different model viruses were treated in bovine serum by two different inactivation methods: samples were subjected either to chemical inactivation with ethylenimine (El) at concentrations of 5 and 10 mM at 37 degrees C for periods up to 72 h or to electron-beam irradiation in frozen and liquid form with doses varying between 11 and 46 kGy. The chemical inactivation resulted in nonlinear tailing curves in a semilogarithmic plot of virus titer versus inactivation time showing non-first-order kinetics with respect to virus titer. The time for inactivation of 7 log10 units of porcine parvovirus (PPV) was about 24 h for both El concentrations, whereas 5 log10 units of bovine viral diarrhea virus (BVDV) was inactivated in 2 h for both El concentrations and 6 log10 units of porcine enterovirus (PEV) was inactivated within 3 h. The inactivation with electron-beam irradiation resulted in almost linear curves in a semilogarithmic plot of virus titer versus irradiation dose, reflecting a first-order inactivation. The rate of inactivation was almost twice as fast in the liquid samples compared to the rate in frozen ones, giving values of the doses needed to reduce virus infectivity 1 log10 unit for inactivation of PPV of 11.8 and 7.7 kGy for frozen and liquid samples, respectively, whereas the corresponding values for BVDV were 4.9 and 2.5 kGy, respectively, and those for PEV were 6.4 and 4.4 kGy, respectively. The nonlinear inactivation with El makes it impossible to extrapolate the curves beyond the virus detection limit and thereby predict the necessary time for complete inactivation, i.e., to a level beyond the detection limit, of virus in a given sample. The first-order inactivation obtained with electron-beam irradiation makes such a prediction possible and justifiable. The two methods are discussed with respect to their different kinetics and applicability under different circumstances and criteria for inactivation, and considerations for choice of method are discussed.

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Ascaris suum: a revision of its early migratory path and implications for human ascariasis.

During the course of carrying out studies on the role of intestinal immunity in blocking the migration of larval Ascaris suum in the pig, it was discovered that the prevailing understanding of larval penetration sites was at variance with our observations. Therefore, a detailed investigation of the migration of A. suum 1.2 larvae through the intestine was initiated. The results demonstrate that the 1.2 larvae invade almost exclusively the wall of the pig cecum and colon and not the small intestine as is generally believed. The larvae were recovered from the mucosa of the cecum and colon as early as 3 hr postinoculation (PI) with infective eggs and were recovered from the liver by 6 hr PI. The maximal recovery of larvae (total larvae and larval/g of mucosa) from the intestinal mucosa occurred between 6 and 12 hr PI; by 24 hr PI, virtually all of the larvae had disappeared from the mucosa. These observations correct a common misunderstanding of this aspect of the life cycle of A. suum in the pig, and they raise 2 issues related to the biology and pathogenesis of Ascaris in humans. What is the actual migratory and development behavior of Ascaris lumbricoides and A. suum in humans and the potential risk for liver lesions? Most authors, in describing the life cycle of A. lumbricoides, either ignore or discount a possible obligatory liver stage of development, and, consequently, the potential for lesion formation similar to that which occurs in pigs infected with A. suum. This issue takes on added importance with the growing evidence that A. suum is an important zoonoses.

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Improved method for the recovery of Ascarus suum larvae from pig intestinal mucosa.

In the course of a comprehensive study on the population biology of Ascaris suum, it became necessary to determine quantitatively the migration of the larvae (L2) through the pig intestine. Because no satisfactory methods for recovering these larvae from the intestinal mucosa were apparently available in the literature, we undertook the development of such a procedure. Direct Baermannization of the intestine proved inadequate, so a series of studies was undertaken to find a method to solubilize the mucosa and free the larvae. Both pepsin digestion and mucus reduction with N-acetyl cysteine were evaluated. The highest recovery rates were obtained with short-term pepsin digestion (45-90 min), followed by a specific settling and washing procedure. A third alternative, migration of larvae out of agar-gel, was also tested. Although the recovery efficiency was low (10% of pepsin digestion), the larvae recovered were clean, with good motility. The agar-gel procedure could be useful when L2 larvae are needed for other purposes, such as antigen preparation or cultivation.

Acetylcysteine↗

The impact of diets varying in carbohydrates resistant to endogenous enzymes and lignin on populations of Ascaris suum and Oesophagostomum dentatum in pigs.

The impact of diets varying in type and level of carbohydrates resistant to endogenous enzymes and lignin on the establishment and location of Ascaris suum and Oesophagostomum dentatum was investigated experimentally. Fifty worm-free pigs, from a specific pathogen-free farm were used. The animals were assigned randomly to 5 diets and infected with 600 infective A. suum eggs and 6000 infective larvae of O. dentatum per pig. The diets consisted of a traditional ground barley plus protein feed (diet A), commercial full-constituent pelleted feed (diet B), barley flour plus protein (diet C), barley flour, inulin (Raftiline ST, ORAFTI, Tienen, Belgium), sugar beet fibre plus protein (diet D), and barley flour, wheat bran, and protein (diet E). The faecal egg excretion was followed and the pigs were slaughtered at 8 weeks p.i. and samples taken from the small and large intestine. Intestinal contents were analysed for worm burdens, worm location and female worm fecundity along with the concentration of insoluble (chromic oxide) and soluble (polyethylene glycol-4000) markers, lignin, non-starch polysaccharides (NSP) and organic acids. In all diet groups A. suum worm burdens were low and comparable, whereas the O. dentatum worm burdens were significantly higher in pigs fed the diets with high levels of NSP and lignin (diets A and E) than in pigs fed diets B, C, and D. The present study suggests that a diet rich in lignin and insoluble NSP's provides favourable conditions for the establishment of O. dentatum in the large intestine of pigs while it is unlikely that the concentration of short-chain fatty acids and pH plays and major role.

Animal Feed↗

Experimental Schistosoma bovis infection in goats: the inflammatory response in the small intestine and liver in various phases of infection and reinfection.

In a histopathological study of goats experimentally infected with Schistosoma bovis, the characteristics of the inflammatory response in the small intestine and liver related to tissue egg counts and fecal egg excretion were compared between goats at different time periods of primary infection and of primary infection followed by challenge. At early patency, coinciding with increasing egg excretion, the intestinal lamina propria showed numerous intact schistosome eggs devoid of any inflammatory reaction, whereas egg-associated inflammatory foci in the intestine were significantly few. Later in primary infection and after challenge, intestinal changes were marked by a granulomatous anti-egg response, with only a minor component of eggs lacking inflammatory change, and were consistent with a reduction of egg transfer into the gut wall. Hepatocellular necrosis with eosinophil infiltration was pronounced only during the early patent stage. The results indicate that the early cascade of fecal egg excretion in caprine schistosomosis bovis is aided by a low degree of tissue reactivity to eggs in the intestine. They also lend support to previous findings indicating that an anti-fecundity effect is operative after exposure to challenge in this parasitic infection in goats.

Animals↗

Nematode-trapping fungi in biological control of Dictyocaulus viviparus.

Larvae of the cattle lungworm Dictyocaulus viviparus were cultured in experimental units of 200 g cattle faeces placed in semi-transparent trays in the laboratory. In each of 4 experimental series using this experimental unit, chlamydospores (chl) of the nematode-trapping fungus Duddingtonia flagrans were admixed to half of the faecal cultures in a concentration of 50.000 chl/g. In all 4 series there was a significant reduction in the development and subsequent release of infective lungworm larvae from faecal cultures containing chlamydospores. The average reduction in larval release, caused by fungal spores, was 86%.

Animals↗

In vivo labeling of Schistosoma japonicum cercariae with 35S.

A technique was developed for in vivo labeling of adult Schistosoma japonicum worms using 35S bound to methionine and cysteine. The isotope was applied in an in vivo labeling system in which Oncomelania hupensis snails harboring a patent S. japonicum were individually exposed to the label. Following exposure to cercariae, mice were perfused after 6 wk. For autoradiography and fluorography, the worms were mounted on cardboard, covered with x-ray films, and exposed for 5 wk. With both techniques, images of both male and female worms were obtained in the 1- and 2-microCi groups, but not in the 0.5-microCi group. Fluorography proved more sensitive than autoradiography. Snail survival, labeled cercariae, and worm establishment were not affected by the radiolabeling. 35S was thus found useful for radiolabeling adult S. japonicum worms.

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Characterization of Oesophagostomum bifurcum and Necator americanus by PCR-RFLP of rDNA.

Esophagostomiasis in humans due to infection with Oesophagostomum bifurcum (nodular worm) is of major human health significance in northern Togo and Ghana, where Necator americanus (human hookworm) also exists at high prevalence. Yet, very little is known about the transmission patterns of O. bifurcum, which is in part due to the difficulties in diagnosis and in differentiating some life-cycle stages of O. bifurcum from N. americanus using morphological features. As a first step toward developing a molecular-diagnostic assay, it was evaluated whether ribosomal (r)DNA could provide genetic markers for the identification of O. bifurcum and N. americanus to species. Internal transcribed spacer rDNA (plus flanking and intervening sequences) was analyzed by polymerase chain reaction-linked restriction fragment length polymorphism (PCR-RFLP) using several restriction endonucleases. The analysis showed that there was no detectable intraspecific difference in the size of the PCR products among multiple samples, that there was a consistent size difference in the products (of 110 bp or 350 bp, depending on region amplified) between the species, and that there was no significant variation in restriction patterns within each species. These results indicate that the rDNA spanning the internal transcribed spacers provides useful genetic markers for the identification of O. bifurcum and N. americanus to species, which has important implications for developing PCR-based tools to study the epidemiology and population biology of O. bifurcum.

Animals↗

Use of an agar-gel technique for large scale application to recover Ascaris suum larvae from intestinal contents of pigs.

Four groups each of 3 pigs were inoculated with Ascaris suum eggs. Pigs in groups 1 and 3 were inoculated with 1000 eggs, and pigs in groups 2 and 4 with 10,000 eggs. On day 10 and 21 post-inoculation (p.i.), respectively, groups 1 + 2 and 3 + 4 were slaughtered, and the contents from the small intestines collected. The contents were mixed with agar to a final concentration of 1% agar and allowed to sediment. The larvae were allowed to migrate from the agar-gel into 38 degrees C 0.9% saline overnight, and were then collected on a sieve (20 microns mesh) and counted. The larvae retained in the agar-gel were counted after pouring the melted agar through a sieve (20 microns mesh). The results showed that more than 97% of the larvae migrated out of the agar-gel and were available for counting in an almost clean suspension. The inoculation dose level did not significantly affect the recovery percentage, neither did the larval stage (10 or 21 days old larvae). The variation in the time interval from slaughtering to start of incubation (interval 57-155 min) did not significantly affect the recovery percentage.

Agar↗

Effect of two virus inactivation methods: electron beam irradiation and binary ethylenimine treatment on determination of reproductive hormones in equine plasma.

Ionizing irradiation and binary ethylenimine treatment have previously been shown to be effective for in-vitro inactivation of virus in biological material. In the present study the 2 methods were tested for possible effects on measurable concentrations of reproductive hormones in equine plasma (luteinizing hormone (LH), folliclestimulating hormone (FSH), progesterone (P4), and oestradiol-17 beta (E2)). The inactivation methods were electron beam irradiation with a dose from 11 to 44 kGy or treatment with binary ethylenimine (BEI) in concentrations of 1 and 5 mmol/L. Generally, there was a close correlation (r > 0.8, < 0.001) between pre- and post-treatment hormone levels. Thus, the different phases of the oestrous cycle could be distinguished on the basis of measured hormone concentrations of treated samples. However, both treatments significantly changed hormone concentrations of the plasma samples. For LH, FSH, and E2 the effect of irradiation and BEI treatment was depressive and dose-dependent. For P4 the effect of irradiation was also depressive and dose-dependent. However, the highest dose of BEI resulted in an increase of measured P4 concentration, which may be attributed to changes in the plasma matrix due to the treatment. Although the treatments affected measured hormone concentrations, the close correlation between pre-treatment and post-treatment measurements means that the diagnostic value will remain unchanged.

Animals↗