Aggregation of Taenia solium cysticerci in pigs: Implications for transmission and control.

Parasite aggregation within hosts is a fundamental feature of parasite distributions, whereby the majority of parasites are harboured by a minority of hosts. Parasite aggregation can influence their transmission and hence control. In this narrative review, possible sources of aggregation of Taenia solium cysticerci in pigs are discussed, along with implications for control of the parasite. While heavy T. solium infections in pigs could most likely be associated with ingestion of high doses of infective parasite eggs, consistent with coprophagic behaviour of pigs, lighter infections indicate a role of indirect routes of transmission to pigs, mostly from lower infection doses. Light infections are likely to be missed by commonly used diagnostic methods - tongue examination or meat inspection - and end up in the food chain. Hence, they entail a 'hidden' risk and are of a particular public health concern, especially in areas where meat is consumed raw or undercooked. To be effective and sustainable, control strategies against T. solium likely require a broader understanding of, and consideration for parasite transmission dynamics. More importantly, a holistic One Health approach incorporating interventions on humans, pigs and the environment will likely have a larger, more successful and sustainable impact.

Trial Design of a Prospective Multicenter Diagnostic Accuracy Study of a Point-of-Care Test for the Detection of Taenia solium Taeniosis and Neurocysticercosis in Hospital-Based Settings in Tanzania.

Taenia solium diagnosis is challenging as trained personnel, good diagnostic tools, and infrastructure is lacking in resource-poor areas. This paper aims to describe the study trial design adopted to evaluate a newly developed rapid point-of-care test that simultaneously detects taeniosis and neurocysticercosis (TS POC) in three district hospitals in Tanzania. The two-stage design included three types of patients: patients with specific neurological signs and symptoms (group 1); patients with complaints compatible with intestinal worm infections (group 2); patients with other symptom(s) (group 3). For group 1, all patients were tested using the TS POC test (stage 1), after which all positive, and a subset of negative, patients were selected for laboratory reference tests, clinical examination, and a brain computed tomography (CT) scan (stage 2). For groups 2 and 3, a similar design was adopted, but clinical examination and a brain CT scan (stage 2) were only performed in patients who were TS POC test-positive for cysticercosis. Due to the lack of a gold standard, a Bayesian approach was used to determine test accuracy for taeniosis and cysticercosis. For neurocysticercosis, a composite case definition was used as the reference standard. If successful, this study will help the future developments (commercialization and implementation) of the rapid test and improve patient management and disease prevention.

Trial Design for a Diagnostic Accuracy Study of a Point-of-Care Test for the Detection of Taenia solium Taeniosis and (Neuro)Cysticercosis in Community Settings of Highly Endemic, Resource-Poor Areas in Zambia: Challenges and Rationale.

Field-applicable, high-quality, and low-cost diagnostic tools are urgently needed for Taenia solium. The aim of this paper is to describe the design, challenges, and rationale for the design of a diagnostic accuracy study in low-resource community settings in Zambia. The trial was designed as a prospective study with a two-stage design to evaluate a new point-of-care test (TS POC) for the detection of taeniosis and (neuro)cysticercosis. Participants within randomly selected households were tested with the TS POC test (index test). Participants who tested TS POC positive for taeniosis and/or cysticercosis and a subset of the negatives were requested to give blood and stool samples for reference testing, and to undergo clinical examination and a cerebral CT scan. The difficulties of conducting a clinical trial in settings with limited research and neuroimaging infrastructure as well as peculiarities specifically related to the disease (low prevalence of taeniosis and the lack of a gold standard) were taken into consideration for the design of this study. The two-stage design increased the efficiency of the study by reducing the number of samples, clinical examinations, and CT scans. Simplified flows and sampling processes were preferred over complex follow-up and randomization systems, aiming to reduce bias and increase the generalizability of the study.

Performance of Ag-ELISA in the diagnosis of Taenia solium cysticercosis in naturally infected pigs in Tanzania.

BACKGROUND: Taenia solium is a zoonotic parasite responsible for neurocysticercosis-a major cause of late-onset acquired epilepsy in humans. Lack of affordable, specific and sensitive diagnostic tools hampers control of the parasite. This study assessed the performance of an antigen detection enzyme-linked immunosorbent assay (Ag-ELISA) in the diagnosis of viable T. solium cysticercosis in naturally infected slaughter-age pigs in an endemic area in Tanzania. METHODS: A total of 350 pigs were bled before they were slaughtered and their carcases examined. Serum was analyzed for circulating antigens by using a monoclonal antibody-based B158/B60 Ag-ELISA. Each carcase was examined for the presence of Taenia hydatigena cysticerci and half carcase musculature together with the whole brain, head muscles, tongue, heart and diaphragm were sliced with fine cuts (< 0.5 cm) to reveal and enumerate T. solium cysticerci. Half carcase dissection can detect at least 84% of infected pigs. Prevalence and their 95% confidence intervals (CI) were calculated in Stata 12. Sensitivity, specificity, predictive values and likelihood ratios were determined. RESULTS: Twenty-nine pigs (8.3%, 95% CI: 5.6-11.7%) had viable T. solium cysticerci while 11 pigs had T. hydatigena cysticerci (3.1%, 95% CI: 1.6-5.5%). No co-infection was observed. Sixty-eight pigs (19.4%, 95% CI: 15.4-20%) tested positive on Ag-ELISA; of these, 24 had T. solium cysticerci and 7 had T. hydatigena cysticerci. Sensitivity and specificity were determined to be 82.7% and 86.3%, respectively. Positive and negative predictive values were 35.2% and 98.2%, respectively. Likelihood ratios for positive and negative Ag-ELISA test results were 6.0 and 0.2, respectively. There was a significant positive correlation between the titre of circulating antigens and intensity of T. solium cysticerci (r(348) = 0.63, P < 0.001). CONCLUSIONS: The Ag-ELISA test characteristics reported in this study indicate that the test is more reliable in ruling out T. solium cysticercosis in pigs, than in confirming it. Hence, a negative result will almost certainly indicate that a pig has no infection, but a positive result should always be interpreted with caution. Estimates of T. solium prevalence based on Ag-ELISA results should, therefore, be adjusted for test performance characteristics and occurrence of T. hydatigena.

TSOL18 vaccine and oxfendazole for control of Taenia solium cysticercosis in pigs: A field trial in endemic areas of Tanzania.

A field trial was conducted in Tanzania to determine the effectiveness of TSOL18 vaccine used concurrently with oxfendazole (OFZ), and of OFZ alone, on T. solium cysticercosis determined by organ and half carcase dissection of slaughter age pigs. This study followed a quasi-experimental group design. Suitable trial sites were randomly allocated to either treatment group T1 (OFZ treatment alone [30mg/kg, Paranthic 10%]) or T2 (TSOL18 [1ml, Cysvax] plus OFZ). Three 4-monthly treatments were administered to eligible pigs. A random selection of pigs were necropsied at baseline and at endline, 2-3.5 months after the final treatment. Additionally, untreated pigs from T1 and T2 areas were necropsied at endline to provide contemporaneous comparisons with T1 and T2 pigs. Baseline prevalence of viable T. solium cysticerci for T1 was 25.5% (Exact 95% CI: 13.9, 40.3; n = 12/47), and for T2 was 12.0% (CI: 6.4, 20.0; n = 12/100). At endline, prevalence was 2.8% for T1 (CI: 0.1, 14.5, n = 1/36) and 0% for T2 (CI: 0, 4.7, n = 0/77). Among untreated pigs, three had viable cysticerci, one from T1 area (12.5%, CI: 0.3, 52.7; n = 1/8) and two from T2 area (5.7%, CI: 0.7, 19.2, n = 2/35). Fisher's exact test showed significant changes in prevalence from baseline to endline in both groups (T1: p = 0.005, T2: p = 0.001). Firth's penalized Maximum Likelihood method suggested the changes were not significant relative to their controls (T1: p = 0.245, T2: p = 0.076). These findings showed a significant reduction in the prevalence of viable cysticerci from baseline to endline after both interventions. However, the changes could not be definitively attributed to the interventions due, in part, to small numbers of control pigs. Concurrent administration of the TSOL18 and OFZ cleared infection among assessed pigs whereas infection remained after treatment with OFZ only. Further studies including larger sample sizes would be required for more definitive conclusions. A One Health approach is recommended for rapid and sustainable impact.

Endemicity of Taenia solium cysticercosis in pigs from Mbeya Rural and Mbozi districts, Tanzania.

BACKGROUND: Taenia solium taeniasis/cysticercosis is a disease of substantial economic and public health importance particularly in low-income countries. The disease was reported to be endemic in Mbeya Rural and Mbozi districts, in the southern highlands of Tanzania, the major pig production area in the country. In 2008, using B158/B60 antigen detection enzyme-linked immunosorbent assay (Ag-ELISA), porcine cysticercosis prevalence of up to 32% was reported in the districts. A number of interventions have been implemented in selected villages including an integrated approach consisting of improving pig confinement and selective treatment of pigs with oxfendazole. Mass drug administration with praziquantel targeting schistosomiasis, with an expected effect on T. solium, was also provided to school-age children in the area. This study aimed at providing an update on prevalence and intensities of porcine cysticercosis; and assessing farmers' knowledge, attitudes and practices which could be associated to disease transmission in the area. The study involved a questionnaire survey conducted using face-to-face household interviews with 890 consenting farmers; and carcass dissections performed on 282 pigs randomly selected from the surveyed households. RESULTS: Twenty-six pigs (9.2%) were infected with T. solium; of which two-thirds (65.4%) had light to moderate infection intensities (1-1000 cysticerci), and one-third (34.6%) had heavy intensities (> 1000 cysticerci). Questionnaire results showed that only 5.7% of the respondents perceived T. solium cysticercosis to be an important disease in pigs. About 18.5% of the respondents were aware of T. solium taeniasis, but 32% of them were unaware of how the infection is acquired. Half of the respondents had seen cysticerci in pork, of whom 61% were not aware that consumption of infected pork could cause taeniasis. Latrines were observed to often (90%) lack doors with 45% considered accessible to pigs. CONCLUSIONS: This study provided an evidence that the disease was still endemic in the area. Poor knowledge of farmers, attitudes, and risky practices responsible for disease perpetuation were also revealed. A One Health approach targeting the whole area incorporating improvement of farmer knowledge regarding disease transmission needs to be trialed as a feasible approach to control.

Hair cortisol and dehydroepiandrosterone concentrations in naturally Taenia solium infected pigs in Tanzania.

The aim of this study was to measure hair cortisol and dehydroepiandrosterone (DHEA) concentrations in naturally Taenia solium infected and non-infected control pigs and assess the effect of an environmental change on the aforementioned parameters. Three hair patches were obtained from 13 T. solium infected and 15 non-infected controls sows, respectively corresponding to 3 time points (prior to, at and approximately two weeks after arrival at the research facility). Cortisol and DHEA were extracted using methanol and analysed by radio immune assay. Mean hair cortisol concentrations were significantly lower (p<0.001) in T. solium infected (4.7±3.0pg/mg) compared to control pigs (9.0±3.7pg/mg) prior to arrival at the research facility, however no significant difference was observed between the two groups at arrival and after approximately two weeks. Similar patterns were also observed for DHEA concentrations (infected pigs 253.9±82.3pg/mg, control pigs 387.7±116.4pg/mg) (p<0.001). Results showed that lean animals had significantly higher cortisol concentrations in both groups, infected and controls pigs, while DHEA was not significantly different between lean and normal animals. Results of this study have shown that an environmental change could have an effect on pigs' hormonal levels suggesting an undergoing adaptation process. After the pigs were kept under the same conditions, fed and watered ad libitum, no significant differences were observed between the groups, but a drop in DHEA concentrations was observed in all the pigs. Weight however had an effect on cortisol levels as lean animals had significantly higher cortisol concentrations in both groups, compared to normal pigs.

Taenia solium in Europe: Still endemic?

The pork tapeworm, Taenia solium, causes an important economic and health burden, mainly in rural or marginalized communities of sub-Saharan Africa, Asia, and Latin-America. Although improved pig rearing conditions seem to have eliminated the parasite in most Western European countries, little is known about the true endemicity status of T. solium throughout Europe. Three recent reviews indicate that autochthonous human T. solium taeniasis/cysticercosis may be possible in Europe, but that current peer-reviewed literature is biased towards Western Europe. Officially reported data on porcine cysticercosis are highly insufficient. Favourable conditions for local T. solium transmission still exist in eastern parts of Europe, although the ongoing integration of the European Union is speeding up modernisation and intensification of the pig sector. Further evidence is urgently needed to fill the gaps on the European T. solium endemicity map. We urge to make human cysticercosis notifiable and to improve the reporting of porcine cysticercosis.

The association between seizures and deposition of collagen in the brain in porcine Taenia solium neurocysticercosis.

Neurocysticercosis caused by infection with Taenia solium is a significant cause of epilepsy and seizures in humans. The aim of this study was to assess the association between seizures and the deposition of collagen in brain tissue in pigs with T. solium neurocysticercosis. In total 78 brain tissue sections from seven pigs were examined histopathologically i.e. two pigs with epileptic seizures and T. solium cysts, four pigs without seizures but with cysts, and one non-infected control pig. Pigs with epileptic seizures had a larger amount of collagen in their brain tissue, showing as large fibrotic scars and moderate amount of collagen deposited around cysts, compared to pigs without seizures and the negative control pig. Our results indicate that collagen is likely to play a considerable part in the pathogenesis of seizures in T. solium neurocysticercosis.

Severe seizures in pigs naturally infected with Taenia solium in Tanzania.

Neurocysticercosis (NCC) caused by Taenia solium is a serious neurological disease. In humans neurological symptoms have been thoroughly studied and documented, however, there is limited information on clinical signs in pigs infected with T. solium cysticerci. Among the scientific community, it is in fact believed that pigs with NCC rarely show neurological signs. The aim of this study was to describe clinical manifestations associated with NCC in pigs and correlate the manifestations to the number and distribution of cysticerci in brains of naturally infected pigs in Tanzania. Sixteen infected and 15 non-infected control pigs were observed for 14 days during daylight hours, and subsequently videotaped for another 14 consecutive days using close circuit television cameras. All occurrences of abnormal behaviour (trembling, twitching, mouth and ear paralysis, ataxia, dribbling, salivating, eye blinking, walking in circles) were recorded. At the end of the recording period, pigs were slaughtered and their brains dissected, cysticerci counted and locations noted. During the recording period, two infected pigs were observed having seizures. Some of the observed autonomic signs during a seizure were chewing motions with foamy salivation and ear stiffening. Motor signs included tonic muscle contractions followed by a sudden diminution in all muscle function leading to collapse of the animal. Stereotypic walking in circles was observed on several occasions. At dissection, both pigs had a high number of brain cysticerci (241 and 247 cysticerci). The two pigs with seizures were also older (36 months) compared to the others (18.3 months, ± 8.2 standard deviation). Results of this study have shown that pigs with NCC can develop clinical signs and suffer from seizures like humans with symptomatic NCC. Results of this study could potentially open up a new experimental pathway to explore the aetiology of neurological symptoms in humans with NCC associated epilepsy.

ADVANZ: Establishing a Pan-African platform for neglected zoonotic disease control through a One Health approach.

Advocacy for neglected zoonotic diseases (ADVANZ) is a One Health Neglected Zoonotic Diseases (NZDs) project, funded by the European Commission through its 7th framework programme. The initiative aims at persuading decision makers and empowering stakeholders at local, regional, and international levels towards a coordinated fight against NZDs. ADVANZ is establishing an African platform to share experiences in the prevention and control of NZDs. The platform will compile and package existing knowledge or data on NZDs and generate evidence-based algorithms for improving surveillance and control with the ultimate aim of eliminating and eradicating these diseases. The platform will serve as a forum for African and international stakeholders, as well as existing One Health and NZD networks and harness and consolidate their efforts in the control and prevention of NZDs. The platform had its first meeting in Johannesburg, South Africa in March 2013.

Dynamics and longevity of maternally-acquired antibodies to Taenia solium in piglets born to naturally infected sows.

This study was designed to investigate the longevity of maternally-acquired specific antibody (Ab) isotypes in piglets born to sows with Taenia solium cysticercosis. Four isotypes of anti-cysticercal immunoglobulins (IgG(total), IgG1, IgG2 and IgA), were assessed in sows naturally infected with T. solium (n=5) and their piglets (5/sow). Specific IgG2 and IgA responses were not detected in piglets at any sampling point. However, IgG(total) and IgG1 responses were detectable in sera from piglets from day 0 but not at 2months of age. Antibody profiles differed significantly over time between littermates and between piglets from different sows. These findings raise important questions as to the optimal timing of mass vaccination of piglets as a method of controlling cysticercosis in humans. The observed variations in the level of maternally-acquired Abs amongst piglets will pose a major challenge to the implementation of vaccination programmes against T. solium cysticercosis.

Real-time polymerase chain reaction for detection of low-intensity Schistosoma japonicum infections in China.

More sensitive methods for diagnosing infection with Schistosoma japonicum are needed as control becomes more effective. We compared a real-time polymerase chain reaction (PCR) for stool samples with conventional diagnostic methods in a study of 1,727 persons from Anhui Province, China. Seroprevalence determined by using an indirect hemagglutination assay (IHA) was much higher (26.1%) than the prevalence in stool-based tests, which were 5.3%, 3.2%, and 3.0% for PCR, hatching test, and Kato-Katz thick smear, respectively. A large proportion of the positive stool samples were only positive in one or two tests. The PCR showed better agreement with IHA than the other two stool-based tests. A commonly used diagnostic algorithm with initial screening for antibodies and subsequent testing with the Kato-Katz thick smear of the seropositive results would have resulted in treatment of 22 people compared with 50 people if the PCR replaced the Kato-Katz thick smear. As prevalence and intensity decrease, the benefit of increased sensitivity using the PCR must be weighed against additional costs.

Low sensitivity of the formol-ethyl acetate sedimentation concentration technique in low-intensity Schistosoma japonicum infections.

BACKGROUND: The endemic countries are in a diagnostic dilemma concerning Schistosoma japonicum with increasing difficulties in diagnosing the infected individuals. The formol-ethyl acetate sedimentation concentration technique is preferred by many clinical microbiology laboratories for the detection of parasites in stool samples. It is potentially more sensitive than the diagnostic methods traditionally used. METHODOLOGY/PRINCIPAL FINDINGS: We evaluated the technique for detection of low-intensity S. japonicum infections in 106 stool samples from China and used a commercial kit, Parasep Midi Faecal Parasite Concentrator. One stool sample and one serum sample were collected from each person. As reference standard we used persons positive by indirect hemagglutination in serum and positive by Kato-Katz thick smear microscopy (three slides from a single stool), and/or the hatching test. We found the sedimentation technique to have a sensitivity of only 28.6% and specificity of 97.4%. CONCLUSION/SIGNIFICANCE: This study indicates that the sedimentation technique has little to offer in the diagnosis of low-intensity S. japonicum infections, at least when only a single stool sample is examined.

Cytokine mRNA profiles in pigs exposed prenatally and postnatally to Schistosoma japonicum.

The pig is a natural host for Schistosoma japonicum and a useful animal model of human infection. The aim of the present study was to assess the differences between the cytokine profiles in prenatally or postnatally S. japonicum exposed pigs. Seven prenatally exposed pigs, 7 postnatally exposed pigs and 4 uninfected control pigs were compared 27 weeks post exposure. Variables included worm burdens, tissue egg counts, liver pathology and mRNA levels of IL-2, IL-4, IL-10, IL-12, TNF-alpha, TGF-beta1 and IFN-gamma in the liver and the caecum, assessed by RT-PCR. Infection intensity and level of septal fibrosis were significantly higher in the postnatal group compared to the prenatal group (P < 0.05). A significant increase of IL-4 (P < 0.01), IL-10 (P < 0.01), IL-12 (P < 0.01) and TNF-alpha (P < 0.05) mRNA level was also observed in the caecum of prenatally infected animals compared to the control group (P < 0.01). The prenatal group showed higher levels of TGF-beta1 in the liver compared with the postnatally infected group (P< 0.05) and the control group (P< 0.01). This suppressive immune response correlated with previously reported low hepatic pathogenesis in prenatally exposed pigs.

Treatment and reinfection of water buffaloes and cattle infected with Schistosoma japonicum in Yangtze River Valley, Anhui province, China.

The aim of the present study was to evaluate the efficacy of praziquantel treatment of Schistosoma japonicum infections in cattle and water buffaloes and to assess the natural rate of reinfection after treatment. The studies were conducted on 2 islands in the Yangtze River, Anhui province, China, from March 2003 to January 2004. The efficacy of praziquantel was 97% when applied orally wrapped in tree leaves at the recommended doses. The efficacy was measured using a miracidium hatching technique on fecal samples collected 20 days after treatment. The treatment did not give rise to any major side effects. Reinfection after treatment was high and occurred throughout the year in both cattle and water buffaloes. Age-related resistance was only observed in water buffaloes. It is concluded that although praziquantel is highly effective against S. japonicum in cattle and water buffaloes, a single annual treatment strategy does not effectively control transmission. New strategies for integrated control of animal schistosomiasis are needed to control schistosomiasis transmission more effectively in farm areas of China.

Distribution of mast cells in relation to Schistosoma japonicum induced lesions in pigs.

The pathogenesis of schistosomiasis japonica has been extensively studied, however only little attention has been paid to the presence and localization of mast cells in relation to Schistosoma japonicum induced lesions. The aim of the present pilot study was to assess the parasitological and pathological responses in S. japonicum infected pigs with emphasis on the description of the distribution of mast cells in relation to lesions in the liver and cecum. Six pigs were exposed to 2,000 cercariae and examined 9 weeks post-infection. Three unexposed pigs of the same age served as helminth free controls. All infected pigs developed granulomatous hepatitis and typhlitis. In the liver, the degree of mast cell infiltration was higher in the infected pigs compared to the unexposed control group. This distinction could not be shown in the cecum. In both the liver and cecum, a mild to moderate number of mast cells were present within the granulomas. A significant relation was found between infection with S. japonicum and the mast cell infiltration in the liver. Due to their possible association with hepatic fibrosis, it seems as if they have some function in the fibrogenic process and thereby play a dual role in the pathogenesis of S. japonicum. In conclusion, the results show that mast cells are recruited to egg induced lesions in both the liver and the cecum.

Schistosoma japonicum eggs survive passage through dogs and chickens.

The present study sought to elucidate whether Schistosoma japonicum eggs could survive a passage through dogs and chickens. Three dogs and three chickens were used for the experiment. The dogs were allowed to eat goat faeces containing 80 S. japonicum eggs per gram. The chickens were given 2 ml of water containing more than 100 S. japonicum eggs per ml. The hatchability of the eggs was 51.8% prior to administration. All faeces were collected for 3 days from the dogs and for 2 days from the chickens and egg excretion and hatchability were measured. The results showed that nearly 50% of the eggs administered to the dogs were recovered in the faeces during the first 2 days and during the first 25 h 39% of the recovered eggs were viable and able to hatch. In the chickens 9% of the administered eggs were recovered within 27 h and within the first 3.5 h after administration 19% of the recovered eggs were viable and able to hatch. The results suggest that both dogs and chickens may act as transport hosts for S. japonicum in endemic areas.

The impact of natural helminth infections and supplementary protein on growth performance of free-range chickens on smallholder farms in El Sauce, Nicaragua.

Three on-farm studies were conducted in Nicaragua during three consecutive years (1999-2001) to assess the impact of natural helminth infections on growth performance of free-range chickens aged 3-4 months. On all participating farms, half of the chickens were treated regularly with anthelmintics (Trifen avicola - a combined formulation of piperazine, phenothiazine and dichlorophen - or albendazole) to express the growth potential of non-infected birds, whereas the other half served as non-treated controls. In 1999, treated chickens had a 39% higher weight gain compared to the control group 6 weeks after the first treatment on 15 farms. In 2000 and 2001, treated chickens had similar weight gain as the control group 10 weeks after the first treatment on 7 farms and 12 farms, respectively. The main reason for the very-different weight gain figures seems to be the weather conditions. In 1999, the study site experienced a rainy season with precipitation far above average, whereas in 2000 and 2001 the rainy seasons had precipitations far below average. Based on these findings, routine use of anthelmintics in the study area would only be recommended in wet years when production losses due to helminth infections seem to be pronounced. In 2001, the study set-up included an assessment of the effect of protein supplementation (soybean) on growth on six farms. Supplemented chickens (treated and non-treated with anthelmintics) had 17% higher weight gain than non-supplemented. Protein supplementation affected neither worm burdens nor faecal egg counts for any of the studied helminths. The post-mortem examinations showed that Trifen reduced burdens of Ascaridia galli, Heterakis gallinarum, and cestodes (efficacies of 100, 100 and 67%, respectively). Albendazole reduced burdens of H. gallinarum (efficacy of 100%). Efficacies against other helminths were difficult to assess due to low worm burdens. Chickens treated with albendazole had lower Ascaridia and Heterakis faecal egg counts than non-treated chickens.