Distribution and density of cysticerci of Taenia solium by muscle groups and organs in naturally infected local finished pigs in Tanzania.

The distribution and density of cysticerci of Taenia solium among distinct carcass sites was determined in 24 naturally infected finished pigs from Mbulu district, Tanzania. The heart, tongue, internal and external masseters, triceps brachii, lungs, liver, kidneys, psoas, diaphragm and brain of each pig as well as the muscles from the forelimb, hind limb, abdomen, head and thorax from one half of each pig carcass were all designated as distinct carcass sites and sliced in such a way that all fully developed cysts could be revealed and enumerated (i.e. each slice was less than 0.5 cm thick). The carcasses harboured from 76 to 80,340 cysts in total. Carcass sites which harboured the highest proportion of cysts were those of the hind and forelimbs (mean: 27.7 and 24.5%, respectively, of the total cysts in the carcass), while lower proportions were found in the tongue, heart, triceps brachii, and diaphragm (7, 3.6, 2 and 2, respectively). Relative cyst density was calculated for the different carcass sites by dividing the mean proportion of the total weight of the tissue groups into the mean proportion of cysts located in that site. The cysticerci in the examined distinct carcass sites were found in the following order of relative density: psoas muscles (10.5), internal masseter (8.1), external masseter (7.1), triceps brachii (4.9), forelimb (4.0), head muscles (3.8), tongue (3.4), hind limb (3.2), diaphragm (2.4), heart (1.9), abdominal muscles (1.3), trunk muscles (1.1), brain (1.0) and oesophagus (0.3). The proportion of cysts expected to be found at the surfaces exposed by visual examination or incision at meat inspection was calculated using an indirect method, which incorporated the area revealed by incision and visual inspection of an organ and the proportion of cysts located in the particular organ. It was estimated that 10.6% of the cysts would be located at inspected sites if regulations were followed carefully.

Schistosoma japonicum infection in the pig as a model for human schistosomiasis japonica.

Valuable information on human schistosomiasis japonica has been provided using primates and experimental rodent hosts. However, major drawbacks such as high costs and ethical concerns for the primate models and large biological deviations for the rodent models have led to the search for more appropriate models. Recent data on the pig indicate that this natural host for Schistosoma japonicum might be a realistic alternative. As only very few research groups have investigated the S. japonicum/pig model, the present review mainly deals with the experimental methods and the major host/parasite findings obtained from the authors own research group. With emphasis on a critical evaluation of the work, the results are compared to the scarce information existing on human schistosomiasis japonica. Like in humans, S. japonicum establishes mainly in the large intestinal veins, with high faecal egg counts during the acute phase of infection, which varies greatly within and between days. Concomitant resistance is another shared feature, but studies in pigs have indicated that the phenomenon is more complex than generally thought. Clinical signs as eosinophilia and diarrhoea with mucus and blood in the acute phase of infection and hepatomegaly, increased portal diameter, periportal fibrosis and ascites in chronic infections are common findings in both humans and pigs. Low protein diet aggravates the disease in pigs by increasing the establishment rates, the faecal egg excretion and the morbidity. A 100% cure rate is achieved when treating S. japonicum infected pigs with praziquantel at 40 mg/kg, and 4 weeks post treatment pigs remain resistant to reinfection. Lastly, human congenital S. japonicum infection has been confirmed in pigs but the implications of such infections for the pathogenesis of schistosomiasis japonica remain to be investigated.

Epidemiology of Echinococcus granulosus in Arbil province, northern Iraq, 1990-1998.

During the period 1990-1998, 99 cases of human cystic hydatidosis (12.4 cases per year) were surgically treated at the two main hospitals in Arbil province, northern Iraq, and from this the human occurence for the province was estimated to be 2 per 100,000 inhabitants. In the same area, 1270 sheep, 550 goats and 320 cattle were examined at slaughter for hydatid cysts and prevalence rates were found to be 15.0%, 6.2% and 10.9%, respectively. A decreasing tendency in livestock prevalences was found towards the end of the study period. As in humans, most of the hydatid cysts in livestock were located in the liver. Fertility of sheep cysts, i.e. those containing protoscoleces, was found to be significantly higher (64%) than that of goats (35.7%) and cattle (29.8%). The percentage of fertile cysts containing viable protoscoleces varied between 63 and 82% in the livers and between 72 and 79% in the lungs of the different animal species. A total of 97 stray dogs were examined post-mortem in the years 1991, 1992 and 1998, and Echinococcus granulosus worms were found in the intestines of 48 dogs (49.5%). High worm burdens (> 1000) were observed in 37% of the dogs, medium worm burdens (200-1000) in 41%, and low worm burdens (< 200) in 22%. In 1998, the prevalence of canine echinococcosis (24.3%) was found to be significantly lower than in 1991 (70.4%) and 1992 (60.6%). The prevalence of human hydatidosis did not differ significantly over the years, but the study confirmed that hydatidosis is endemic in northern Iraq, and that housewives, labourers and farmers appear to be at the greatest risk of infection.

Isolation and characterisation of sex-specific transcripts from Oesophagostomum dentatum by RNA arbitrarily-primed PCR.

In light of the lack of molecular data on the sexual differentiation, maturation and interaction of parasitic nematodes of livestock, the present study investigated sex-specific gene expression in the nodule worm, Oesophagostomum dentatum (Strongylida). Using the technique of RNA arbitrarily-primed polymerase chain reaction (RAP-PCR), 31 expressed sequence tags (ESTs) differentially-displayed between the sexes were cloned. Northern blot analysis proved ten ESTs to be expressed exclusively in males (adults and fourth-stage larvae), while two were expressed solely in female stages. None of the ESTs were expressed in infective third-stage larvae. Sequence analysis and subsequent database searches revealed two male-specific ESTs to have significant similarity to Caenorhabditis elegans (predicted) proteins, a protein containing an EGF-like cysteine motif and a serine/threonine phosphatase. Another two male-specific ESTs had similarity to non-nematode sequences. The two female-specific ESTs had similarity to vitellogenin-5 and endonuclease III (predicted) from C. elegans. The remaining ESTs had no similarity to any nucleic acid or protein sequences contained in the databases. The isolation and characterisation of sex-specific ESTs from O. dentatum provides a unique opportunity for studying the reproductive biology of parasitic nematodes at the molecular level, with a view toward novel approaches for parasite control.

Visceral larva migrans: migratory pattern of Toxocara canis in pigs.

The migratory pattern of Toxocara canis was investigated following infection of pigs with 60000 infective eggs. Groups of six pigs were slaughtered at 7, 14 and 28 days after infection (p.i.), and the number of larvae in selected organs and muscles was determined by digestion. A group of uninfected pigs was used as negative controls for blood parameters and weight gain. Toxocara canis migrated well in the pig, although the relative numbers of larvae recovered decreased significantly during the experiment. On day 7 p.i., high numbers of larvae were recovered from the lymph nodes around the small intestine and to some extent also from the lymph nodes around the large intestine, and from the lungs and the liver. On day 14, the majority of larvae were recovered from the lungs and the lymph nodes around the small intestine, and by day 28 p.i. most larvae were found in the lungs. Larvae were recovered from the brain on days 14 and 21, with a maximum on day 14 p.i. No larvae were found in the eyes. Severe pathological changes were observed in the liver and lungs, especially on day 14 p.i.; also, development of granulomas was observed in the kidneys. Finally, a strong specific antibody response towards T. canis L2/L3 ES products was observed from day 14 p.i. until termination of the experiment, and the maximum eosinophil response was observed 14 days p.i. The pig is a useful non-primate model for human visceral larva migrans, since T. canis migrate well and induce a strong immunological response in the pig. However, the importance of the pig as a paratenic host is probably minor, because of the relatively early death of most of the larvae.

An immunohistological study of phenotypic characteristics of cells of the inflammatory response in the intestine of Schistosoma bovis-infected goats.

The cellular inflammatory response in the small intestine of 21 goats infected with Schistosoma bovis was phenotypically characterized by immunohistochemistry between 6 and 32 weeks post-exposure, with particular reference to perioval granulomatous reactions. Macrophages of granulomas consistently expressed MHC class II molecules, whereas multi-nucleated giant cells in general did not. Most granulomas contained moderate infiltrates of CD2+ (CD4+ or CD8+) and gamma/delta (T19+) T cells, whereas B lymphocytes were sparse. Intact extravascular mucosal eggs, lacking appreciable cellular reactivity on plain histopathology, displayed surrounding collars of MHC class II+ macrophages. Gamma/delta T cells and MHC class II+ macrophages were the predominant cell types in perivascular inflammatory cell clusters in the submucosa. The phenotypic cellular composition of granulomas did not change appreciably with duration of infection. The results indicate the importance of MHC class II-restricted immune events in the caprine S. bovis egg granulomas and also suggest a role of gamma/delta T cells in their pathogenesis. It is hypothesized that the early appearance of perioval macrophage collars may serve to protect eggs from ovicidal host defence mechanisms, facilitating excretion and continuation of the life-cycle.

Parasites in cross-bred pigs in the Upper East region of Ghana.

A cross-sectional study was carried out in the Upper East Region (UER) of Ghana in order to estimate the prevalence of parasitic infections in local cross-bred pigs. Out of 60 villages with a human population of 200-1000 inhabitants, 10 villages were randomly selected for the study. The number of pigs varied from 50 to 200 pigs per village. In total 259 faecal samples from growers were collected and examined. Ninety-one percent of the animals excreted parasite eggs. Among these the prevalence of Eimeria spp. was 77.2%, Isospora suis (27%) and Balantidium coli (19.3%).The following helminth eggs were identified: Metastrongylus salmi (19.3%); Physocephalus sexalatus (17.4%); Oesophagostomum spp./Hyostrongylus rubidus (60.6%); Trichuris suis (4.6%); Ascaris suum (12.7%); Ascarops strongylina (8.1%); Brachylaemus suis (1.9%); Paragonimus suis (0.8%); Globocephalus urosubulatus (2.7%); and Schistosoma suis (0.4%). Furthermore, six growers were selected from each village for clinical and postmortem examinations, i.e. 60 in total. The clinical examinations revealed ectoparasites on 98.3% of the animals. The ectoparasites were: Haematopinus suis (66.7%); Boophilus spp. (58.3%); Amblyomma spp (45.0%); Sarcoptes suis (38.3%); and Rhipicephalus spp. (8.3%). All pigs were examined for the presence of haemoparasites. It was found that 23.3% of the animals had haemoparasites. These were: Babesia perroncitoi (23.3%); Babesia trautmanni (13.3%); and Eperytrozoon suis (1.7%). Based on postmortem examinations the following adult worms were identified: Metastrongylus salmi (83.3%); Oesophagostomum dentatum (63.3%); Oesophagostomum quadrispinulatum (38.3%); Hyostrongylus rubidus (23.3%); Ascarops strongylina (76.7%); Globocephalus urosubulatus (20.0%); Strongyloides spp. (1.7%); and Physocephalus sexalatus (65.0%). Cysts of the human tapeworm Taenia solium, Cysticercus cellulosae, were present in 11.7% of the animals. Small pieces of the diaphragm were examined for the presence of Sarcocystis spp.. The prevalence was 28.3%, but no larvae of Trichinella spp. were found. Furthermore, four of the animals (6.7%) had Taenia hydatigena cysts.

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.

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.

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.

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.

Experimental single and trickle infections of cattle with Taenia saginata: studies of immunodiagnosis.

Two groups of 12 calves were given either a single infection dose of 30,000 Taenia saginata eggs (group A) or trickle infected with 12 x 2500 T saginata eggs (group B). The calves were killed 16, 22 or 54 weeks after infection and the numbers of viable and dead parasites were counted by slicing selected organs and muscles of half of the carcases. The total numbers of cysts and the numbers of viable cysts were higher in group A than in group B. Within the two groups there were no significant differences between the numbers of cysts at the three dates of slaughter. Comparable results were obtained by using an ELISA and a dot blot procedure for the detection of antibody in the serum of the infected calves; an initial increase in response during the first 10 weeks after infection was generally followed by a gradual decline. When using a monoclonal antibody-based ELISA for the detection of circulating antigens, the sera from all the 14 animals which had no viable cysts or only dead cysts at slaughter did not react with the monoclonal antibodies. Calves 122 and 123, which harboured two and 122 viable cysts, respectively, reacted positively during the late phase of the infection to the antigen-detecting ELISA. However, several other animals which harboured between two and 41 viable cysts at slaughter did not react.

Immunodiagnosis of Taenia saginata in cattle using hydrophobic antigens from T. hydatigena metacestode cyst fluid.

Hydrophobic fractions isolated from cyst fluid of Taenia hydatigena metacestodes, obtained from naturally infected goats from Tanzania, were used in an indirect ELISA and in immunoblot (Western or dot blot) procedures. The use of the most hydrophobic fractions of the cyst fluid proved to result in a better diagnostic discrimination in ELISA than the more hydrophilic fractions. Best diagnostic ratio between responses from T. saginata infected and non-infected cattle was achieved when using horse-radish peroxidase-labelled sheep antibovine IgG1 as the conjugate. When the hydrophobic fractions of T. hydatigena cyst fluid were employed as antigen in ELISA, no cross-reaction took place with sera from cattle infected with Fasciola hepatica, Ostertagia ostertagi, Cooperia oncophora, Dictyocaulus viviparus, Sarcocystis cruzi or a mixture of D. viviparus, Trichostrongylus axei and Eimeria. Using the Western blot technique, specific low molecular weight components (M(r) 10-18 kDa) of the hydrophobic fractions of the cyst fluid were reactive with sera from 14 of the 21 calves infected with T. saginata for 60 days. Also, hydrophobic fractions of T. hydatigena cyst fluid were dotted onto strips of filter membranes and reacted specifically with sera from 21 calves infected with T. saginata. This technique resulted in the highest sensitivity, detecting 17 of the 21 calves infected with T. saginata. Calves harbouring as few as 16, 26, 43, 53 and 74 cysts could be detected. Thirteen calves harbouring between 74 and 2,545 cysts were all detected. However, 4 calves harbouring 2, 22, 52 and 66 cysts, respectively, were not detected. The potential of using a dot blot technique in large scale investigations in the tropics is present. However, before accepting any of the three tests in the tropical environment, a broader spectrum of helminth infections should be evaluated for their eventual cross-reactivity.

The prevalence of Taenia solium metacestodes in pigs in northern Tanzania.

Eighty three carcasses of pigs were examined at three abattoirs in Moshi, Arusha and Mbulu in northern Tanzania. Taenia solium metacestodes were found in all the three abattoirs with an overall prevalence of 13.3%. During routine meat inspection in Kiboroloni, Moshi 6.2-6.9% of the pigs were found to harbour T. solium metacestodes. The mean number of hooks on the protoscolices was 27 and the length of the small hooks varied from 105 microns to 130 microns while that of the larger hooks varied from 168 microns to 174 microns confirming that the cysts were metacestodes of T. solium. Although cystlike lesions were recovered from livers of both pigs and cattle, no hooklets of sizes in the range of 5-37 microns were found indicating that no Taiwan Taenia metacestodes were recovered during carcass examination.

Ability of the fungus Pleurotus pulmonarius to immobilise preparasitic nematode larvae.

In vitro experiments showed that the oyster mushroom Pleurotus pulmonarius exerts a strong immobilising effect on preinfective larvae of the animal parasitic nematodes Ostertagia ostertagi, Cooperia oncophora, Oesophagostomum quadrispinulatum and Cyathostoma species. Infective stages of the same species seemed less vulnerable to fungal attack, possibly because they are protected by their outer cuticle.

Evaluation of an enzyme-linked immunosorbent assay (ELISA) for detection of Taenia saginata cysticercosis in cattle.

Serum IgG response of cattle with cysticercosis caused by Taenia saginata was studied in an enzyme-linked immunosorbent assay (ELISA) where a T. saginata metacestode surface extract was used as antigen. In experimentally infected calves, a sharp rise in specific antibody levels was found 3-4 weeks after the infection followed by a logical level of detection corresponded to about 25 cysts. The ELISA was employed in cattle herds where cysticercosis outbreaks had occurred and also in supposedly uninfected herds. Significantly increased antibody levels were found in the herds with massive cysticercosis cases. The test was not adapted for individual diagnosis as some animals of the uninfected herds, especially within the older age groups, had elevated antibody values. The ELISA was, however, useful in the investigation of outbreaks to determine the extent and pattern of the infection in the herd. The rate of decline in antibody levels in these herds was studied by follow up sampling. The increased antibody levels in the infected herds were also reflected in colostrum-fed calves. This observation was employed to estimate the time of infection.

A case-control study of risk factors in light Taenia saginata cysticercosis in Danish cattle.

Risk factors in light T. saginata cysticercosis in cattle herds were investigated in a case-control study. The case group consisted of farms from which cattle with low-grade cysticercosis had been detected at slaughter. The control group consisted of farms where no cases were detected during the same period. The major risk factor identified was allowing cattle access to drink from streams carrying effluent from sewage treatment plants (odds ratio = 3.6). Spreading of septic tank sludge through deliberate or accidental mixing with animal slurry, deposition of sewage sludge, and proximity to railways, camping sites or sewage treatment plants seemed to be of minor importance.

Distribution of Taenia saginata cysts in carcases of experimentally infected calves and its significance for routine meat inspection.

The distribution of Taenia saginata cysts was recorded in 23 experimentally infected calves. The calves harboured between two and 2569 cysts. A median of 15.7 per cent of the cysts were located in the heart and 6.5 per cent in the masseter muscles, the organs usually considered as the most important predilection sites. From these findings a simple probability model was used to estimate the meat inspection sensitivity at different infection levels. It was concluded that the possibility of detecting infected cattle is limited especially when, as in Denmark, the majority of animals are lightly infected.

Bovine cysticercosis in Denmark. A study of possible causes of infection in farms with heavily infected animals.

Epidemiological studies were made on 14 farms from which, during a 2 year period, 38 cattle had been condemned at slaughter, due to massive infections with Cysticercus bovis. By on-site investigations and interviews, attempts were made to identify the transmission routes of Taenia saginata eggs from human faeces into the environment and further on to cattle. The most frequent sources of infection were found to be sludge from septic tanks illegally applied on pasture or crops, in some cases after having been mixed with animal slurry. Animals in permanently housed herds were infected through the fodder or by contamination of the indoor environment by such slurry containing Taenia eggs. Other herds were infected by grazing pastures in close proximity to municipal sewage treatment plants. In contrast to earlier Danish observations, application on farmland of sewage sludge from municipal treatment plants was not involved in any of the reported outbreaks. This apparent change coincides with the implementation of more restrictive legislation for the agricultural use of sewage sludge in Denmark.

A study on the survival of Taenia saginata eggs on soil in Denmark.

The infectivity of Taenia saginata eggs exposed to environmental conditions on a natural soil surface in Denmark was studied by feeding the eggs to susceptible calves, followed by determination of the number of cysts developed. The results indicated that a small proportion of the eggs remained infective for 6 1/2 months, but not for 9 1/2 months when deposited in May 1986, and for 5 1/2 months but not for 8 1/2 months when deposited in September 1987. Viability of eggs was tested in vitro and compared with infectivity obtained in calves.