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Brucellosis is an important but neglected zoonosis that causes serious economic losses both in livestock and human populations. The aim of the present study was to estimate the true prevalence of brucellosis together with diagnostic sensitivity and specificity of three serological tests in humans of the northwestern part of Ecuador using a Bayesian approach adjusted for the dependencies among the multiple tests to avoid any misinterpretation. In addition, the causal agent responsible for human brucellosis was also identified. Using a total of 3,733 samples collected from humans in this area between 2006 and 2008, the prevalence of human brucellosis and the diagnostic test characteristics of the Rose Bengal fast agglutination test (RBT), Wright’s slow agglutination test with ethylenediaminetetraacetic acid disodium salt dehydrate (EDTA) (SAT-EDTA), and indirect ELISA (iELISA) were estimated using a Bayesian approach. The estimated true prevalence of human brucellosis was 1% (credibility interval: 0.4–1.6). The sensitivities of iELISA and RBT were higher than and similar (95.1% and 95.0%, respectively) to those of SAT-EDTA (60.8%). Even though all tests indicated a high specificity (> 99.0%), the specificity of SAT-EDTA was highest (99.9%). The circulating strain in this study area was identified to be Brucella abortus biotype 4 based on culture and microbiological characterization. The RBT and the iELISA are recommended for estimating the true prevalence of human brucellosis and/or for surveillance programs following their high sensitivities and specificities. The proposed strategy supports evidence-based medicine for clinicians and policy-makers to ensure appropriate preventive and control program of brucellosis worldwide.

We present a 44-year-old man from a rural community in northern Ecuador who worked on a cattle farm where he was involved with primary veterinary care, including assistance during births (or calving) and placenta retention and artificial insemination, with minimal precautions. In September of 2009, quite abruptly, he developed asthenia and hypersomnia without any apparent cause or symptoms like fever, chills, or night sweats. On November 14, 2009, he suffered from pain and edema in the right testicle that coincided with pain in the abdomen. Clinical, serological, and bacteriological investigations confirmed the first case of unilateral orchitis in man in Ecuador caused by Brucella abortus biovar 1. Because brucellosis is a neglected disease, special attention should be given to it in the training of medical and veterinary students.

The aim of the present study was to develop a PCR-ELISA assay for the detection and differentiation of the main African pathogen trypanosomal species present in peripheral blood of cattle. The proposed methodology allows to specifically differentiate Trypanosoma congolense , Trypanosoma vivax and the subgenus Trypanozoon , by means of a sensitive universal PCR amplifying trypanosome DNA followed by an ELISA-based hybridization with three highly specific probes. The semi-nested PCR had a sensitivity of 15 fg, 15 fg, and 0.15 fg of DNA from T. vivax , T. congolense , and Trypanosoma brucei brucei , respectively that is sufficient to detect parasites in blood during the chronic phase of the disease. Biotinylated second round asymmetric PCR amplification products were used in an ELISA set up using three species-specific probes for the diagnosis of T. congolense (type Riverine, Kilifi or Savannah), T. vivax and T. brucei brucei . A factor O.D. of 0.082 was determined on blood samples from bovines ( n = 18) from a non-endemic area in Africa. In a pilot study of blood samples of naturally and experimentally Trypanosoma infected cattle previously characterized by PCR-RFLP ( n = 42), a high rate of concordance (93.3%) was found between PCR-RFLP and PCR-ELISA. There is a good ratio between positive and negative O.D. values (3.00 vs. 0.1) and the technique can also be used to distinguish mixed infections.

Background. Neurocysticercosis (NCC) is a frequent cause of epilepsy worldwide. Compared with the more common parenchymal brain cysts, extraparenchymal infections are difficult to manage and have a poor prognosis. Serological assays are used to detect circulating Taenia solium antigens or anti-T. solium antibodies in serum or cerebrospinal fluid (CSF) samples. There are no guidelines on whether to use serum or CSF specimens for a particular assay. Methods. We obtained paired serum and CSF samples from 91 patients with NCC (48 had intraparenchymal NCC, and 43 had extraparenchymal NCC) for detection of antibodies, using an enzyme-linked immunotransfer blot (EITB) assay, and antigens, using a monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA). Results. For the intraparenchymal NCC group, the EITB assay yielded more true-positive results for serum samples, and the ELISA yielded slightly more true-positive results for CSF samples than for serum samples, but none of these differences were statistically significant. Most patients with calcified NCC were antibody positive but antigen negative. For extraparenchymal disease, all samples were antibody positive, and all but 2 were antigen positive, with most samples containing high antigen levels. Conclusions. The sensitivity of antibody-detecting EITB assays is not increased through the use of CSF samples rather than serum samples. The antigen-detecting ELISA performed better for CSF samples than for serum samples, but for both specimen types it was less sensitive than the EITB assay. Active and inactive NCC are better differentiated from each other by the antigen-detecting ELISA, for both serum and CSF samples. High antigen levels suggest the presence of subarachnoid NCC.

Neurocysticercosis (NCC) is a major cause of seizures and epilepsy. Diagnosis is based on brain imaging, supported by immunodiagnosis in serum or cerebrospinal fluid (CSF). Lumbar puncture is invasive and painful. Blood sampling is slightly painful and poorly accepted. Urine antigen detection has been used for other parasites and tried in NCC with suboptimal performance. We used a monoclonal antibody-based ELISA to detect Taenia solium antigens in urine from 87 Peruvian neurocysticercosis patients (viable cysts, N = 34; subarachnoid cysticercosis, N = 10; degenerating parasites, N = 7; calcified lesions, N = 36) and 32 volunteers from a non-endemic area of Peru. Overall sensitivity of urine antigen detection for viable parasites was 92%, which decreased to 62.5% in patients with a single cyst. Most patients (30/36, 83%) with only calcified cysticercosis were urine antigen negative. Antigen levels in paired serum/urine samples (evaluated in 19 patients) were strongly correlated. Non-invasive urine testing for T. solium antigens provides a useful alternative for NCC diagnosis.

A survey on porcine trichinellosis was organised in Ecuador between 2000 and 2003. Blood samples were taken in slaughterhouses (study 1, n = 2000; study 2, n = 331) and in a remote village where pigs are free roaming (study 3, n = 646) and examined by ELISA using excretory/secretory (E/S) antigens. Seven samples (0.35%) in study 1 and none of the samples of study 2 were serologically positive. Thirty-seven (5.72%) village pigs tested positive by E/S ELISA in study 3. Sero-positive results by the E/S ELISA in study 1 were confirmed by ELISA using β-tyvelose antigen, and by immunoblot. Muscle samples taken from pigs slaughtered in the abattoir (study 2) and from animals that showed a positive serology in study 3 were examined by trichinoscopy and artificial digestion. These techniques failed to demonstrate the presence of muscle larvae. The results of this survey need confirmation, but suggest that Trichinella is present in Ecuador; however, prevalence and parasite burdens are likely to be very low. The likelihood of detecting trichinellosis are higher in traditional settings than in pigs raised on improved farms.

Several diagnostic techniques are used to estimate the prevalence of the zoonotic tapeworm Taenia solium in pigs, but none of these tests are perfect, making interpretation of results difficult. A Bayesian approach was used to estimate values for the prevalence and diagnostic test characteristic of porcine cysticercosis by combining results of four imperfect tests. Village pigs (N = 868), slaughtered in Lusaka (Zambia), were bled, and tongue and routine meat inspected-, and serum antibody and parasite antigen concentrations were determined by ELISA. A model. based on a multinomial distribution and including all possible interactions between the individual tests required 3 1 parameters to be estimated, but actually allowed only 15 parameters (i.e. had 15 degrees of freedom) to be estimated. Therefore, prior expert opinion on specificity and (in)-dependence of the tests was entered in the model, resulting in a reduction of the number of parameters to be estimated. 9 The estimated prevalence of porcine cysticercosis was 0.642 (95% confidence interval 0.54-0.91). The performances of the tests were (sensitivity (se) -specificity (sp)): tongue inspection (se 0.210-sp 1.000), meat inspection (se 0.221 -sp 1.000), Ab-ELISA (se 0.358-sp 0.917), Ag-ELISA (se 0.867-sp 0.947). To validate the estimates obtained from the model we performed a second study: 65 randomly purchased Zambian village pigs were bled for serum antibody and antigen determination, their tongue and meat inspected; and in addition, the carcasses were dissected for total cysticercus counts (gold standard). Cysticerci were found in 3 1 pigs (prevalence 0.477, 95% confidence interval 0.35-0.60). overlapping with the estimated prevalence in the first stud. Sensitivity and specificity values obtained for they aforementioned tests in this study were in agreement with those estimated. A Bayesian analysis framework offers the possibility to combine prior opinion with experimental data to more accurately estimate the real prevalence of porcine cysticercosis in the absence of a gold standard. (C) 2003 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

In West Africa, Taenia solium cysticercosis in both pigs and man has been reported in Benin, Burkina-Faso, Ghana, Ivory Coast, Senegal and Togo, and although official data are lacking, T. solium is anticipated to be present in most of the pig-raising regions of other West African countries as well. In some regions of Nigeria, the prevalence of porcine cysticercosis and human taeniosis is quite high (20.5 and 8.6%, respectively). Surprisingly, however, no cases of human cysticercosis have been reported, although epilepsy is very common. Large epidemiological surveys have only been carried out in Togo and Benin, where the prevalence of human cysticercosis was 2.4 and 1.3%, respectively. In Central Africa, porcine and human cysticercosis are (hyper)-endemic in Rwanda, Burundi, the Democratic Republic of Congo and Cameroon. The parasite also has been reported in pigs in Chad and Angola. Cysticercosis has been shown to be one of the major causes of epilepsy in Cameroon with figures as high as 44.6%. Cameroon is one of the few countries where the taeniosis-cysticercosis complex has been examined more in detail. In the Western province of Cameroon large scale surveys have shown that active cysticercosis is present in 0.4-3% of the local population and in 11% of the village pigs. However, the prevalence of adult T. solium was only 0.1%, which underscores the frequency of the T. solium paradox. Based on the available information, a very conservative economic estimate indicates that the annual losses due to porcine cysticercosis in 10 West and Central African countries amount to about 25 million Euro. The financial losses due to human cysticercosis are very difficult to estimate, but are certainly exceeded by the social impact of the disease, especially because of the particular perception of epilepsy in many African communities. It is concluded that the true prevalence of T. solium cysticercosis in pigs and humans in Central and West Africa remains underestimated because of unreliable slaughterhouse data and the lack of awareness and diagnostic facilities in the public health sector.

The development of improved immunodiagnostic tools has contributed to our knowledge on the importance of taeniosis/cysticercosis by enabling sero-epidemiological surveys and community-based studies to be carried out. Immunodiagnostic techniques include detection methods for specific antibodies and for circulating parasite antigen in serum or cerebrospinal fluid. The antigens used in immunoblot and enzyme-linked immunosorbent assay (ELISA) for antibody detection have evolved from crude extracts to highly purified specific fractions and recombinant antigens of the glycoprotein family, increasing both the sensitivity and the specificity of the tests. The application of ELISA for the detection of circulating parasite antigens may present some diagnostic advantages since it demonstrates not only exposure but also active infections. Until now only a few of the current techniques have been standardised and fully validated, making comparisons between studies difficult. The lack of a gold standard is a serious drawback. In surveys on cysticercosis, antibody detection systems have been useful in identifying the risk factors associated with transmission of Taenia solium; a high seroprevalence in a community indicates a "hot spot" where preventive and control measures should be applied. In contrast, the potential use of immunodiagnostic tools to identify cases of neurocysticercosis (NCC) in man is subject to debate. The correlation between a positive serology and neurological symptoms and/or lesions indicative for NCC on neuro-imaging techniques is poor to fair in most studies. This may be explained by the unpredictable clinical outcome of the infection and the variable immunological response of the human host to infection. A major problem is that in many developing countries, neuro-imaging methods are inaccessible and/or too expensive for the rural population at risk. Under these conditions, serology may provide the only tool for diagnosis of the infection.

The definitive version is available at www3.interscience.wiley.com, We studied the occurrence of human cysticercosis in 4993 individuals from three rural communities of Menoua Division, West Province of Cameroon. Circulating antigens of Taenia solium metacestodes were detected in 0.4%, 1.0% and 3.0% of the serum samples taken in Bafou, Bamendou and Fonakekeu, respectively, and examined using a monoclonal antibody-based enzyme-linked immunosorbent assay. This test detects only carriers of living cysticerci and gives thus a good idea of the presence of active cysticercosis. The percentage of persons infected with cysticercosis increased with age. Twenty-two of the 34 seropositives underwent computed tomography (CT) of the brain. Thirteen of them were CT-scan positive, which shows that neurocysticercosis was present in 59.1% of the tested seropositive persons. No living cysticerci were detected among 20 seronegative people. About 20.6% of the seropositives had a history of or current taeniasis against only 1.9% of the seronegatives. Based on these figures and on the data on porcine cysticercosis (prevalence: 11%) and human taeniasis (prevalence: 0.13%) collected in the same region, we conclude that T. solium cysticercosis is an endemic, but overlooked public health problem in West Cameroon.

Contrary to the numerous reports on methicillin-resistant Staphylococcus aureus (MRSA) in domestic animals, only three articles concerning zoo animals are documented in the literature. A skin infection of an African elephant (Loxodonta africana) calf was most likely acquired from an infected caretaker. Another zoo detected MRSA in the rumen content of a mouflon (Ovis aries), and, in a third facility, it was reported in a fistulous wound at the coronary band of a digit of an Indian rhinoceros (Rhinoceros unicornis). In the present study, which lasted 13 months and involved 93 different individual mammals that belonged to 40 species and 19 families housed in the Royal Zoological Society of Antwerp, Belgium, this study reports the absence of MRSA in swabs of nostrils, skins, conjunctiva, vulva, abscess, and arm rests in public spaces. Samples were enriched overnight and inoculated on a selective chromogenic medium.

A survey to monitor the use of trypanocidal drugs by cattle breeders was conducted in Zambia. Use was made of a questionnaire and of the isometamidium-ELISA technique. One hundred and twenty-two farmers and 50 veterinary assistants were interviewed. The isometamidium-ELISA was used to monitor the isometamidium serum concentration in 72 cattle, 1 week after unsupervised treatment by 56 farmers and 16 veterinary assistants. Although there was no clear indication of underestimation of the weight of the animals and although farmers had adequate knowledge of the correct usage of isometamidium, the results suggest frequent underdosing when considering isometamidium serum concentrations 1 week after treatment. In 76% of the cases, the expected protection period was equal or shorter than 28 days and equal or shorter than 33 days in 90% of the treated cattle.

Distribution data for epidemiologically important ticks (Acari: Ixodidae) in the Southern Province of Zambia, one of the main cattle areas of the country, are presented. Boophilus microplus (Canestrini) was not recorded in southern Zambia, whereas Boophilus decoloratus (Koch) is present throughout the area. New distribution patterns for less economically important ixodid ticks are also discussed. Southern Zambia is a transition zone because it is the most northern area in Africa where mixed Rhipicephalus appendiculatus Neumann and Rhipicephalus zambeziensis Walker, Norval & Corwin populations were reported. Although a second generation of adult R. appendiculatus/R. zambeziensis was encountered, simulations indicated that this phenomenon is very rare in southern Zambia, mainly because of the colder temperatures during the early dry season and lower rainfall. These simulations were supported by a development trial under experimental conditions. Tick body size measurements showed that southern Zambian ticks are larger than eastern Zambian R. appendiculatus . It is hypothesized that body size is related to diapausing intensity in this species. The epidemiological consequences are that a different approach to control Theileria parva (Theiler) (Piroplasmida: Theileriidae) and other tick-borne diseases is needed in southern Zambia, compared to the one adopted in eastern Zambia.

La presente etude a ete conduite d'aout 1999 a avril 2000 et avait pour objectif de determiner la prevalence de la taeniose due a Taenia solium dans Bafou et Bamendou, deux groupements villageois de la Menoua (Ouest-Cameroun). Quatre (0,13 %) sur 3 109 echantillons de matieres fecales humaines ont ete trouves positifs pour des oeufs de Taenia spp. par la technique de la flottaison. Trois des quatre vers expulses etaient des T. solium et un etait T. saginata . Deux cas de cysticercose etaient presents dans une des familles ou vivait un porteur de T. solium .En plus, des enquetes coprologique et serologique pour la taeniose et la cysticercose a T. solium ont ete faites chez un groupe a risque constitue de bouchers et/ou langueyeurs (n = 137). Les resultats ont ete compares a ceux d'un groupe temoin (n = 198). Aucun porteur de Taenia n'a ete detecte par l'examen microscopique. Les prevalences de la cysticercose dans les deux groupes ont ete respectivement 3,6 et 4,5 %.

To determine the prevalence of porcine cysticercosis, a survey was carried out in 27 villages belonging to two rural communities of West-Cameroon (Bafou and Bamendou). Between January and August, 2000, a total of 707 pigs were examined serologically and by tongue inspection. Serum samples were examined for circulating parasite antigen using a monoclonal antibody-based sandwich enzyme-linked-immunosorbent assay (Ag-ELISA) and for antibodies against cysticerci (Ab-ELISA). Seventy eight samples (11.0%) were found positive in the Ag-ELISA and 154 (21.8%) in the Ab-ELISA, while by tongue inspection on the same animals cysticerci were detected in 43 pigs (6.1%). Gibbs sampling using results of these three tests indicated that the estimated prevalence of porcine cysticercosis was 10.9%. Analysis of the Ag-ELISA results demonstrated that adult pigs showed a significantly higher seroprevalence (15%) than young ones (8.4%). There was no statistical difference in cysticercosis prevalence in pigs raised in households with or without a latrine. Animals that were reported to be usually confined were significantly less infected (9.9%) than free-roaming pigs (16.2%). Infection rates were significantly higher in pigs that had access to human faeces (13.8%) than those which did not have access (9.1%). This study has identified some community behavioural and environmental practices that should be modified to prevent continuous transmission of porcine cysticercosis. (C) 2002 Elsevier Science B.V. All rights reserved.

Diapause in adults of the African brown ear tick Rhipicephalus appendiculatus is dependent on latitude and stock origin. Lower latitude populations, that is, those nearer the equator, show no diapause response to long or short day exposure. Diapause response to short daylengths is found in higher latitude populations, further south, and increases with increasing latitude. Nearly all individuals in the southernmost population, used in this study, enter diapause under short and long day exposure which implies that the onset of diapause in this population is obligatory. Diapause termination in higher latitude populations is induced by increasing photoperiod, further north, but still south of the equator, diapause is terminated by ageing. The differences in diapausing behaviour of the different populations are discussed in relation to their climatic conditions.

Glucosuria in okapis (Okapia johnstoni) was first documented in 1980, yet the etiology remains unclear. In August 2006, an attempt to lower glucosuria in captive okapi by diet modification (omitting all fruit and adding unmolassed beet pulp) was started at the Antwerp Zoo. To study the possible relationship between glucosuria and diet, stress, and/or pregnancy, four okapis were monitored over a period of 4.5 yr. One animal, born in 2006, became glucosuric near the age of three. Three okapis were adults at the start of the study and had been glucosuric for more than 5 yr. The glucose/creatinine urinary ratio values of these four glucosuric animals did not change considerably over time despite dietary changes. Stress did not appear to influence glucosuria in these okapi. Urinary ratio decreased during the second half of pregnancy in two females. In conclusion, the diet change did not reduce glucosuria, but pregnancy appeared to lower urinary glucose in okapis.

Subarachnoid neurocysticercosis carries poor prognosis and high mortality. Besides neuroimaging, there are no available monitoring tools. We examined antigen levels in sequential serum samples of a patient who required four courses of antiparasitic treatment. Antigen levels decreased in parallel to parasite resolution. Antigen detection may contribute to monitoring treatment of subarachnoid neurocysticercosis.

Two trials were carried out in order to compare the prophylactic effect of a subcutaneously implanted sustained release device (SRD) containing a mixture of a biodegradable copolymer, poly(caprolactone-co-L-lactide), and isometamidium (ISMM) with that obtained after intramuscular injection of the drug. In a first experiment under controlled conditions, two groups of cattle were treated with 0.5 mg/kg isometamidium either as a SRD or intramuscularly (i.m.), and exposed at monthly intervals to Glossina morsitans morsitans infected with Trypanosoma congolense. The average protection period was at least 24 months in the SRD treated against 5.7 months in the i.m. treated group. Using an ISMM enzyme-linked immunosorbent assay, the drug could be detected until 140 days post-treatment in the latter group, whereas in the former group, traces of the drug were detectable until 330 days after treatment. Furthermore, a field trial was carried out at the Madina Diassa ranch in Mall involving three groups of N'Dama cattle, each containing 23 or 24 animals. Two groups were treated with 1 mg/kg ISMM either as a SRD or i.m. and a third group served as untreated control. Twelve months after treatment, the cumulative infection rates were 56.5, 87.8 and 91.6% in the SRD implanted, the i.m. treated and the control groups, respectively. The ISMM concentrations were slightly lower than in the laboratory trial, but the overall pattern of drug disappearance from the sera of the SRD treated cattle was very similar in both trials. Statistical analysis showed that the incidence of trypanosomiasis was significantly lower in the SRD treated than in the i.m. treated group. (C) 1999 Elsevier Science B.V. All rights reserved.

Newly-moulted adults of three Rhipicephalus appendiculatus stocks, originating from Kenya, Zambia and Zimbabwe, were kept in different photoperiods (12h : 12h, 13h : 11h and 14h : 10h light : dark). The stock originating from Kenya showed almost no behavioural diapause in any of these day lengths, whereas virtually all individuals of the Zimbabwean stock entered a behavioural diapause irrespective of day length. Adults from the Eastern Province of Zambia, a transition zone between the multivoltine phenology in equatorial Africa and the univoltine in southern Africa, exhibited a photoperiod-dependent diapause response. The possible mechanisms of diapause regulation and their origin are discussed.

The results of tick surveys carried out in the Eastern province of Zambia between December 1982 and February 1996 were principally in agreement with the findings of earlier surveys conducted during the period 1965-72. Boophilus decoloratus has almost been replaced by Boophilus microplus. Hyalomma truncatum was found in small numbers throughout the province and Hyalomma marginatum rufipes was only rarely encountered in collections made from cattle. Rhipicephalus evertsi evertsi was largely confined to the southern part of the plateau and the valleys of the Zambezi tributaries. The main difference between the present survey and the previous one concerns the status of Rhipicephalus appendiculatus. This species is currently expanding its range in a westward direction, whereas it was virtually absent from the southern part of the province during the period 1965-72. The majority of specimens collected are morphologically intermediate between R. appendiculatus and the closely allied Rhipicephalus zambeziensis. The available evidence indicates that R. zambeziensis (sensu stricto) is absent from the province. The phenology of R. appendiculatus is aberrant in the province: at lower altitudes a second generation of adult ticks is recorded on the hosts at the start of the dry season.

Although previous epidemiological surveys in Ecuador indicate the presence of Entamoeba histolytica, prevalence data of this parasite remain scarce. Most of the studies were based on microscopic examination, which does not allow a morphological differentiation from the non-pathogenic Ent. dispar and Ent. moshkovskii. In the present study, 674 stool samples from a South Ecuadorian rural community were screened for Entamoeba spp. Subsequently, molecular identification was performed on 101 samples containing Ent. histolytica/Ent. dispar/Ent. moshkovskii cysts. The study indicated the absence of Ent. histolytica in this South Ecuadorian community and confirmed the difficulty of differentiating Entamoeba spp. based on morphological features.

A series of monoclonal antibodies (MoAb) produced against excretory and secretory products from 10- and 20-week-old Taenia saginata cysticerci were tested for their ability to detect circulating antigen in a double antibody sandwich enzymelinked immunosorbent assay (ELISA). Two MoAb, 12G5 and 2H8, proved to be highly reactive with the tegument of viable T. saginata cysticerci and recognized antigenic components of 65, 87 and 100 kDa in immunoblotting. The detection limit of the assay using 12G5 as trapping antibody and 2H8 as a biotinylated indicator antibody was 0.1 ng protein per ml. Although the sensitivity of the test varied from one animal to another, the minimum number of living cysticerci, which could be detected by the ELISA, was 88. Animals harbouring only dead cysticerci gave similar reactions as non-infected control animals. Cross-reactions were only observed with taeniid parasites. The test was able to detect circulating antigen also in sheep and pigs, respectively infected with T. ovis and T. solium and in the serum samples of confirmed cases of human T. solium cysticercosis.

Four calves were infected with a susceptible (laboratory) strain of Ostertagia ostertagi and four with a field strain suspected to be resistant to benzimidazoles. After 25 days two calves from each group were treated with 3.5 mg kg−1 fenbendazole. Egg output fell to zero in all treated calves. Treated calves did not harbour worms at slaughter 35 days after infection. Significant differences between the strains were shown for ED50 values for thiabendazole in the egg hatch assay, but not in the tubulin binding assay. It is concluded that benzimidazole resistance in the suspected strain cannot be confirmed.

Neurocysticercosis is recognised as an important but neglected cause of epilepsy in developing countries where the parasite occurs. Data on the transmission dynamics of the parasite in endemic areas are scarce. Individuals living in these areas are likely to be highly exposed to the parasite, but relatively few of them develop active infections. The present study aimed to describe and gain insights into changes in antibody responses and infection patterns related to age and/or gender in a south Ecuadorian rural population by combining antibody and antigen serological data with demographic characteristics. In 25% of the population, antibodies to Taenia solium cysticerci were detected whilst 2.9% had circulating parasite antigens. The proportion of antibody-positive individuals increased significantly until the age of 40years to become stable in older individuals. A rule-based simulation model was developed to explain these variations and to reflect the dynamics of exposure to, and transmission of, the parasite. In contrast, the proportion of people presenting circulating parasite antigens, reflecting an active infection, was significantly higher in people older than 60years. Immunosenescence could explain such an observation since a weaker immune system in the elderly would facilitate the establishment and maintenance of viable cysticerci compared with fully immunocompetent younger individuals. This work points out the role of the immune system in the development of cysticercosis within an exposed population and highlights new essential issues in understanding the transmission dynamics of the parasite, its incidence and the resulting immunological response at a population level. Copyright 2009 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

Theileria parva sporozoite stabilates are used in the infection and treatment method of immunization, a widely accepted control option for East Coast fever in cattle. T. parva sporozoites are extracted from infected adult Rhipicephalus appendiculatus ticks either manually, using a pestle and a mortar, or by use of an electric homogenizer. A comparison of the two methods as a function of stabilate infectivity has never been documented. This study was designed to provide a quantitative comparison of stabilates produced by the two methods. The approach was to prepare batches of stabilate by both methods and then subject them to in vitro titration. Equivalence testing was then performed on the average effective doses (ED). The ratio of infective sporozoites yielded by the two methods was found to be 1·14 in favour of the manually ground stabilate with an upper limit of the 95% confidence interval equal to 1·3. We conclude that the choice of method rests more on costs, available infrastructure and standardization than on which method produces a richer sporozoite stabilate.

Molecular markers, especially surface markers associated with type II, cytokine-dependent, alternatively activated macrophages (aaMF), remain scarce. Besides the earlier documented markers, macrophage mannose receptor and arginase 1, we demonstrated recently that murine aaMF are characterized by increased expression of found in inflammatory zone 1 (FIZZ1) and the secretory lectin Ym. We now document that expression of the two members of the mouse macrophage galactose-type C-type lectin gene family (mMGL1 and mMGL2) is induced in diverse populations of aaMF, including peritoneal macrophages elicited during infection with the protozoan Trypanosoma brucei brucei or the Helminth Taenia crassiceps and alveolar macrophages elicited in a mouse model of allergic asthma. In addition, we demonstrate that in vitro, interleukin-4 (IL-4) and IL-13 up-regulate mMGL1 and mMGL2 expression and that in vivo, induction of mMGL1 and mMGL2 is dependent on IL-4 receptor signaling. Moreover, we show that expression of MGL on human monocytes is also up-regulated by IL-4. Hence, macrophage galactose-type C-type lectins represent novel surface markers for murine and human aaMF.

A new method of diagnosing cysticercus or larval stage of the human tapeworm, Taenia saginata, also known as Cysticercus bovis, in formalin-fixed bovine tissue was developed using a monoclonal antibody to T. saginata and avidin–biotin complex immunohistochemistry. Grossly recognizable viable and degenerate cysts were identifiable after immunohistochemical staining and could be differentiated from Sarcocystis, Actinobacillus, or non-cyst, normal bovine structures. The new test should permit laboratory confirmation of suspected T. saginata cysticercus lesions.

A survey to investigate resistance to drugs used in the treatment of bovine trypanosomosis was conducted in the eastern province of Zambia between 1996 and 1998. A cross-sectional study was conducted in three districts (Petauke, Katete, Lundazi) at 34 village sampling sites selected at random from villages that had shown greater than 6% prevalence of bovine trypanosomosis during an earlier survey. A longitudinal study was conducted in same three districts over a 1-year period. The study sites were chosen from the cross-sectional study and included eight sites showing high trypanosomosis prevalence and where no control activities were recorded. Use was made of parasitological methods, tests of resistance in cattle and mice and isometamidium-ELISA. Overall mean prevalence of trypanosomosis was 14.4, with 96% of infections caused by Trypanosoma congolense. The remainder was caused by Trypanosoma vivax (2%) and Trypanosoma brucei (2%). Tests in mice showed that of the stabilates collected, 24 (34%) were resistant to only isometamidium chloride, 8 (11.3%) were resistant to only diminazene aceturate, 1 (1.4%) was resistant to both drugs, and 38 (53.5%) were sensitive to both drugs. At least 2 out of 27 stabilates tested in cattle appeared to be resistant to trypanocidal drugs, 1 to isometamidium and 1 to diminazene. Isometamidium could be detected in only 63 (4.1%) of 1526 serum samples from cattle in the study. Only 6 (2.8%) of 212 serum samples from trypanosome-infected cattle had serum levels of the drug above 0.4 ng isometamidium per ml serum which is indicative for drug resistance in the infecting parasite population. Although some drug resistance is apparent, diminazene aceturate and isometamidium chloride can still be expected to be effective as a sanative pair in this area in most cases, since not more than 1 stabilate of 71 investigated showed evidence of resistance to both drugs.

The first record of the tapeworm Echinococcus multilocularis (Cestoda, Taeniidae) in Red foxes (Vulpes vulpes) in northern Belgium is described. Between 1996 and 1999, 237 dead foxes were examined for the presence of this tapeworm using the intestinal scraping technique. Four foxes (1.7%) were found to be infected with E. multilocularis and showed medium to very high parasitic burdens. Three infected foxes originated from the south of the study area and the fourth animal came from the north of the study area near the border with The Netherlands. These findings are discussed in relation to the high endemicity of E. multilocularis in southern Belgium and to the increased distribution of the Red fox (V. vulpes) in northern Belgium during the last two decades.

In vitro studies were focussed on the duration and cessation of merogony in Theileria parva infected blood lymphocyte cell cultures. The cultures were infected using purified tick stabilates as an alternative to in vitro infections, using sporozoites obtained by labour intensive dissections of salivary glands from infected ticks. After establishment of infection in peripheral blood lymphocytes (PBL), merozoites were temporarily produced for about 2 months after which lymphoblasts only contained schizonts.

During January 2002 the frequency of Taenia solium cysticercosis was studied in a series of 504 epileptic patients from 3 rural localities in the West and North-West provinces of Cameroon using an enzyme-linked immunosorbent assay for both circulating antigen (Ag-ELISA) and antibody (Ag-ELISA) detection. Taenia solium antigens were detected in the sera of 1.2% of the patients whereas specific antibodies against the parasite were present in 44.6% of the patients. Significantly less seropositive results in Ab-ELISA were recorded in Batibo than in Bandjoun and Bamendjou, whereas a borderline significant difference was recorded with increasing age. Furthermore, 56.2% of patients with late-onset epilepsy showed antibodies against cysticercosis. Taenia solium cysticercosis appears to be an important cause of epilepsy in Cameroon.

Seven patients with active neurocysticercosis (NCC) received an eight days treatment with albendazole and were followed up using computed tomography (CT-scan) and a monoclonal antibody based ELISA for the detection of circulating antigen (Ag-ELISA). Only three patients were cured as was shown by CT-scan and by the disappearance of circulating antigens one month after treatment. After a second course of albendazole therapy, two other patients became seronegative. CT-scan showed the disappearance of viable cysts in all persons who became seronegative whereas patients who were not cured remained seropositive. These preliminary results show that this Ag-ELISA is a promising technique for monitoring the success of treatment of NCC patients because of the excellent correlation between the presence of circulating antigens and of viable brain cysts.

Diapause in adults of the African brown ear tick Rhipicephalus appendiculatus is dependent on latitude and stock origin. Lower latitude populations, that is, those nearer the equator, show no diapause response to long or short day exposure. Diapause response to short daylengths is found in higher latitude populations, further south, and increases with increasing latitude. Nearly all individuals in the southen most population, used in this study, enter diapause under short and long day exposure which implies that the onset of diapause in this population is obligatory. Diapause termination in higher latitude populations is induced by increasing photoperiod, further north, but still south of the equator, diapause is terminated by ageing. The differences in diapausing behaviour of the different populations are discussed in relation to their climatic conditions.

A study was carried out in two villages and one marketplace of the Batibo sub-division in North-West Cameroon to determine the prevalence of porcine cysticercosis. The results showed that 4.44% of 383 pigs were positive at tongue examination whereas ELISA detected circulating antigens in 27.7% of 271 pig sera. A questionnaire survey in 140 pig-raising households indicated that 59.3% of them lacked latrines while in 75.7% of the households members defecated directly into pigpens. The seroprevalence of porcine cysticercosis was significantly higher in households without latrines than in those with latrines. Similarly, significantly more seropositive pigs were present in households that defecated in the pig pens (35.5%) than in those that did not (14.4%). Although 91.4% of pig raising households did know of pig cysticercosis, only 28.6% were aware of the link with human taeniasis and only 10.7% were aware of human cysticercosis.

An enzyme-linked immunosorbent assay (ELISA) for detecting circulating Taenia solium antigen was evaluated in Viet Nam; 12 of 210 people gave a positive result, including 5 persons with epilepsy. Cysticercosis was confirmed in 9 persons. Agreement between the ELISA, computerized tomography scanning and biopsy examination was high.

Une étude menée de mai à octobre 1999 dans le département du Mayo- Danay (extrême nord du Cameroun) et dans la préfecture du Mayo-Kebbi (sud-ouest du Tchad) a eu pour but de déterminer la prévalence de la cysticercose porcine et d’identifier les principaux facteurs qui la favorisaient. Elle a montré que les conditions hygiéniques dans lesquelles vivait la population ainsi que celles de l’élevage des porcs étaient très médiocres, avec pour conséquence des infestations massives de la population porcine par des cysticerques de Taenia solium. En effet, 42 p. 100 des 126 exploitations visitées étaient dépourvues de latrines et les porcs, en divagation permanente ou semi-permanente, avaient facilement accès aux matières fécales humaines déposées aux alentours des habitations. Le diagnostic clinique effectué par la méthode du langueyage a montré que 20,5 p. 100 des 852 porcs vivants examinés étaient porteurs de cysticerques. L’inspection des carcasses réalisée dans les abattoirs locaux a révélé que 15,7 p. 100 des 51 porcs abattus étaient ladres. Parmi les 264 sérums de porcs soumis au test Elisa pour la détection des antigènes circulants de cysticerques, 105 (39,8 p. 100) se sont révélés positifs. Ces résultats ont indiqué que le Mayo-Danay et le Mayo-Kebbi étaient d’importants foyers de la cysticercose porcine à Taenia solium.

Belgian Landrace piglets were experimentally infected with eggs of a Taenia sp. of Korean origin. At autopsy, metacestodes were present only in the livers. The proportion of degenerated metacestodes increased from 12%-39% at 5 weeks to 94%-100% at 10 weeks after infection. A sandwich enzyme-linked immunosorbent assay (ELISA) using monoclonal antibodies raised against the excretory-secretory products of T. saginata metacestodes detected circulating antigen in the sera of the pigs at 1 week post-infection. A good correlation was found between the presence of viable metacestodes and the detection of circulating antigen; the latter disappeared as the metacestodes died off. However, the antibodies were detected only after 3 weeks of infection and onwards until the necropsy of the pigs.

Results of a longitudinal study conducted in the eastern province of Zambia from 1994 to 1997 indicate that it is doubtful whether a state of endemic stability of East Coast fever (ECF) can be reached in the near future. Even in endemic areas, the mortality of Theileria parva infections is still estimated above 50%. The main factors limiting progress towards endemic stability are high innate susceptibility of the Zebu cattle, the virulence of the parasite and the climate. The unimodal rainfall pattern results in restricted activity of Rhipicephalus appendiculatus instars and year-to-year variation in rainfall causes fluctuations in tick phenology and T. parva transmission. Adult tick activity invariably peaks during the rains and is associated with the highest ECF incidence. Nymphal transmission of T. parva to cattle appears to be less important. Second periods of activity of both adult and nymphal instars are pronounced only when the climate is suitable. These second waves of tick activity ensure a more continuous and efficient transmission of T. parva and also play a key role in the dynamics of prolonged outbreaks in epidemic areas. ECF control methods may have an important influence on ECF epidemiology. Immunizations as well as chemotherapy of clinical cases create a reservoir of virulent parasites in susceptible cattle, resulting in artificial endemic stability.

An overview is presented of the results obtained with biodegradable sustained release devices (SRDs) containing a mixture of polymers and either isometamidium (ISMM) or ethidium. Under controlled laboratory conditions (monthly challenge with tsetse flies infected with Trypanosoma congolense) the protection period in SRD treated cattle could be extended by a factor 2.8 (for ethidium) up to 4.2 (for ISMM) as compared to animals treated intramuscularly with the same drugs. Using a competitive drug ELISA ISMM concentrations were detected up to 330 days after the implantation of the SRDs, whereas after i.m. injection the drug was no longer present three to four months post treatment. Two field trials carried out in Mali under heavy tsetse challenge showed that the cumulative infection rate was significantly lower in the ISMM-SRD implanted cattle than in those which received ISMM intramuscularly. Using ethidium SRD, however, contradictory results were obtained in field trials in Zambia and in Mali. The potential advantages and inconvenients of the use of SRDs are discussed and suggestions are made in order to further improve the currently available devices.

just beginning to understand the genetics of speciation’. We believe that the current abuse of the species concepts confuses the evolutionary biology debate and we want to clarify this confusion. There are three elements of concern regarding the species concept: (i) taxonomy; (ii) evolution; and (iii) biological diversity assessment. The species as a taxonomic unit is not a real problem as long as keys for species recognition are available to the public. Describing evolution and its mechanisms, or assessing biological diversity are rendered difficult by the use of any species concept. Applying a species concept within the context of evolutionary biology forces one to deal with the evolution of a fixed entity. The evaluation of biological diversity cannot be performed properly if any type of species concept is used. This was demonstrated with our bacteria and birds example in Ref. [1]. Alternative tools for evaluating biological diversity that could be generalized are being developed [2]. The Escherichia coli lineage provides one example that illustrates these problems. The name E. coli is used to describe commensal bacteria of humans that protect the digestive tract from other microbes, whereas the name Shigella spp. describes another group of bacteria that are deadly pathogens of humans. Physicians consider E. coli and Shigella spp. as two different genera because of the differences in their effects on human health. However, Shigella are actually forms of E. coli that recurrently appear by lateral transfers of genes between different E. coli strains [3]. Thus, the distinction between E. coli and Shigella is clinically useful, but it is not helpful in terms of evolutionary biology or biodiversity assessment.

The diapausing behaviour of Rhipicephalus appendiculatus was studied under quasi-natural conditions in the Eastern Province of Zambia. Newly-moulted adults of ticks indigenous to the study area entered a behavioural diapause when exposed to daylengths below a critical photoperiod, provisionally a daylength between 11 h 20min and 11 h 45min. In the Eastern Province of Zambia the diapause was apparently not terminated by a long-day signal, but by a weakening of the photoperiodic maintenance of the diapause because of increasing age of the ticks. Adults of a reference stock indigenous to Kenya also entered a diapause when exposed to daylengths below the same threshold and maintained this diapause for the same length of time. Adults of a reference stock of Rhipicephalus zambeziensis indigenous to Zimbabwe did not enter a diapause when exposed to the same daylengths. The relevance of the findings is discussed in relation to the distributions of the two species.

Four monoclonal antibodies (MoAbs) produced against the excretory-secretory (ES) antigens of Taenia saginata metacestodes are currently used for the detection of circulating antigen in the sera of cattle with bovine cysticercosis. The epitopes recognised by these MoAb were characterized by periodate oxidation, alkaline borohydride and heat treatment, trichloracetic acid precipitation, enzymatic proteolysis and deglycosylation of the ES-products. These results together with those of the homologous sandwich, inhibition and competition ELISAs showed that three repetitive epitopes are recognized, which are present on the tegument and on the ES-products of T. saginata metacestodes. They are different from phosphorylcholine and seem to be either of carbohydrate or partly carbohydrate/partly protein nature.

A monoclonal antibody-based dot-ELISA was developed for the detection of antigen in the sera of cattle with Taenia saginata cysticercosis. After pretreatment of the sera with trichloroacetic acid, antigen could be detected in experimentally infected animals from five weeks after infection. The assay distinguished animals harbouring living cysts from those carrying only dead cysts. A reliable diagnosis of animals harbouring less than 100 cysticerci was not possible, but in cattle with more than 100 living cysticerci the sensitivity was 87·5 per cent; the specificity of the test was 93·5 per cent. These values were very similar to those obtained with a sandwich ELISA.

In parallel with the Th1/Th2 dichotomy, macrophages are capable of developing into functionally and molecularly distinct subpopulations, due to differences in, for example cytokine environment and pathological conditions. While the best-studied, classically activated macrophage is induced by type I stimuli such as IFN-γ, a type II cytokine environment antagonizes the classical activation of macrophages and is capable of alternatively activating macrophages. However, molecular markers associated with these type II cytokine-dependent, alternatively activated macrophages remain scarce. Besides the earlier documented markers macrophage mannose receptor and arginase 1, we recently demonstrated that murine alternatively activated macrophages are characterized by increased expression of FIZZ1 and Ym. We now report that expression of the two members of the mouse macrophage galactose-type C-type lectin gene family, termed mMGL1 and mMGL2, is induced in diverse populations of alternatively activated macrophages, including peritoneal macrophages elicited during infection with the protozoan Trypanosoma brucei or the Helminth Taenia crassiceps, and alveolar macrophages elicited in a mouse model of allergic asthma. We also demonstrate that, in vitro, interleukin-4 and interleukin-13 upregulate mMGL1 and mMGL2 expression and that, in vivo, induction of mMGL1 and mMGL2 is dependent on interleukin-4 receptor signalling. Moreover, we show that regulation of MGL expression is similar in human monocytes and monocyte-derived macrophages. Hence, macrophage galactose-type C-type lectins represent novel markers for both murine and human alternatively activated macrophages; thus, paving the way for further characterization of the phenotype of macrophages occurring in Th2 conditions.

Given the constraints of classical diagnostic methods, i.e., morphological and isoenzymatic studies of proglottids, a polymerase chain reaction test complemented with restriction enzyme analysis has been modified by redesigning one of the primers to reduce nonspecific amplifications experienced when using field samples. The use of these new, highly cestode-specific primers and the restriction enzyme Ddel led to the development of a diagnostic assay that clearly distinguishes between Taenia saginata and T. solium proglottids in field samples. This assay confirms the presence of T. saginata in Ecuador. DNA amplification of some of these taeniids showed different patterns, suggesting the possibility that strain differences exist. These results demonstrate the need for development of useful molecular assays as reliable tools for epidemiological studies on cestodes.

In order to study the reliability of the larval paralysis test as an in vitro assay for the detection of resistance of nematodes to levamisole and morantel tartrate, the influence of different parameters was evaluated using resistant and susceptible Ostertagia ostertagi strains. The operator, the sample (10% of the larvae present in the suspension), the incubation time (24, 48 or 72 h), the incubation temperature (20 or 25 degrees C) and the observation period of the larvae (5 or 15 s) had no statistically significant influence on the test results. Statistical differences were obtained only when L3 larvae of different ages (1 or 2 months) were used. Reversibility of the paralysis did not occur when concentrations of levamisole of less than or equal to 200 micrograms ml-1 or morantel tartrate of less than 2000 micrograms ml-1 were used. The reproducibility of the test was fairly good, with a mean standard deviation of 21.3% for the LC50 values. Morantel resistance in a strain of O. ostertagi was not confirmed as such by the larval paralysis test. The resistance factor was less than or equal to 1, in spite of an efficacy of morantel of 77.4% as shown in vivo.

Jirds (Meriones unguiculatus) were treated with dexamethasone and inoculated subcutaneously with 100–300 oncospheres ofTaenia saginata. Ten of the jirds received 0.5 mg of the drug per animal twice weekly during the first month after inoculation and then once a week until necropsy and four of these became infected. Up to 20 viable metacestodes were recovered in the subcutaneous tissue at the site of inoculation three months after infection.

Wouters G., Geerts S., Brandt J., Kumar V. and Calus A. 1988. Meriones unguiculatus as an experimental host for Taenia saginata metacestodes. International Journal for Parasitology 18 : 129–131. Three hundred and eighty-eight jirds were inoculated subcutaneously with hatched and activated oncospheres from 24 different batches of Taenia saginata . Metacestodes were recovered in 5.2% of the inoculated animals, though significant differences in the infectivity of individual batches were observed. The dose of infection inoculum, the rate of oncospheral activation, the period of egg storage and immunosuppressive treatment of the host also influenced the results. In those harbouring metacestodes, an average of 2.7 viable cysts perjird were recovered at the subcutaneous site of inoculation.

Observations on a beef cattle farm in Flanders led to the suspicion of resistance to levamisole in a strain of Ostertagia ostertagi. After treating a group of six animals with levamisole (5 mg kg-1 L.W., i.m.) the reduction in the number of trichostrongylid eggs per gram of faeces varied between 0 and 66.6%, whereas a similar group treated with fenbendazole (7.5 mg kg-1 L.W., p.o.) showed a reduction in worm burdens of 100%. Coproculture showed that the remaining eggs in the first treatment group were all Ostertagia sp. The suspected field strain was compared with a reference strain of O. ostertagi by means of the in vitro larval paralysis test. This test showed LC95 values of 9.12 micrograms ml-1 and of 99.04 micrograms ml-1 for the reference and the field strain respectively, which indicates a resistance factor for the latter of 10.9. These results were not unequivocally confirmed by the post mortem findings on a tracer calf necropsied 4 days after treatment with levamisole.