Comparison of Kato-Katz, PCR and coproantigen for the diagnosis of Taenia solium taeniasis.

Four methods were compared for the diagnosis of human taeniasis caused by Taenia solium. Fecal samples from persons living in a T. solium endemic region of Madagascar were examined for taeniid eggs by the Kato­Katz method. Subsequently, samples positive (n = 16) and negative (n = 200) for T. solium eggs were examined by (i) amplification of the fragment of small subunit of the mitochondrial ribosomal RNA (rrnS) gene using conventional polymerase chain reaction (PCR) and (ii) a nested PCR of a fragment of the T. solium Tso31 gene. Additionally, 12 egg-positive and all egg-negative samples were tested for coproantigen detection. A further 9 egg-positive fecal samples were examined using both PCRs. Of the 12 egg-positive samples tested by PCRs and coproantigen methods, 9 (75%) were positive by rrnS PCR, 3 (25%) using Tso31-nested PCR and 9 (75%) by coproantigen testing. None of the 200 egg-negative fecal samples was positive in either rrnS or Tso31-nested PCR. Twenty of the 25 egg-positive samples (80%) were positive in rrnS PCR, and DNA sequencing of PCR amplicons was obtained from 18 samples, all confirmed to be T. solium. Twelve of the 25 egg-positive samples (48%) were positive in the Tso31-nested PCR, all of which were also positive by rrnS PCR. It is suggested that species-specific diagnosis of T. solium taeniasis may be achieved by either coprological examination to detect eggs or coproantigen testing, followed by rrnS PCR and DNA sequencing to confirm the tapeworm species in egg-positive or coproantigen-positive samples.

Confirmation by necropsy of a high prevalence of porcine cysticercosis in a rural district of Madagascar.

Neurocysticercosis is recognized as an important health issue in the Malagasy population. To date, investigations into prevalence of infection with the causative agent, Taenia solium, in the parasite's natural animal intermediate hosts, have relied on serological methods which have been found to be non-specific. We determined the prevalence of porcine cysticercosis among pigs from a contiguous area of the Betafo and Mandoto administrative districts, Vakinankaratra Region, Madagascar. One hundred and four slaughter-weight pigs were examined by detailed necropsy examination including slicing of the heart, tongue, masseter muscles, diaphragm and carcase musculature. Thirty-seven animals (35.6%) were found infected with T. solium, representing one of the highest rates of infection ever reported, worldwide. These findings highlight the importance of T. solium in Madagascar and support the need for increased efforts to prevent the parasite's transmission to reduce its burden on the health of the Malagasy population.

How could an African swine fever outbreak evolve in an enzootic context? The case of Imerintsiatosika, Madagascar in 2015.

African swine fever (ASF) is a haemorrhagic contagious pig disease generally causing high mortality. ASF is enzootic in Madagascar with outbreaks reported each year. An ASF outbreak occurred in May 2015 in the municipality of Imerintsiatosika in Madagascar. We investigated the outbreak to describe it and to identify risk factors in order to propose control measures, and to document evidence of an ASF outbreak in an enzootic country. We took biological samples from very sick and dying pigs, sold by the farmer to the butcher, for PCR analysis. An active search for all possible farm-cases was carried out. A definition of suspected farm-case was established and we implemented a descriptive survey and a retrospective cohort study. Laboratory results confirmed ASF virus infection. Suspected farm-cases represented 81 farms out of 922. Out of 3081 pigs of infected farms, 44% (95% CI: 42-46%) were sick, of which 47% were sold or slaughtered. Case fatality was 60% (95% CI: 56-63%) while 21% (95% CI: 19-24%) of the diseased pigs recovered. The outbreak duration was nine months and half of the infected farms' pig population remained after the outbreak. Compared to the exotic breed, local pigs had twice the risk of infection. It is the first detailed report of an ASF outbreak in an enzootic situation. The disease still has a large impact with 50% animals lost. However, the case fatality is lower than expected that suggests the possibility of resistance and subclinical cases. Proximity to road and increased number of farms are risk factors so biosecurity measures are needed. Further studies are needed to understand why pigs of local breed are more affected. Finally, an acceptable alternative to the sale of sick animals should be found as this currently is the breeders' means to reducing economic loss.

Bio-guided identification of proteins for the diagnosis of cysticercosis in swine.

Neurocysticercosis (NCC) is one of the most prevalent parasitic infection of the brain and the most common cause of seizures in adults in tropical countries. Cysticercosis is caused by larvae of Taenia solium, a human tapeworm. Pig or humans are infected by ingestion of eggs in food contaminated by human feces. Diagnosis and treatment of pigs is a pillar of the control of the disease in a country. However current diagnostic tests are based on ELISA and/or Western blot using native antigens needing laboratory facilities not available in rural areas. Development of a pen side diagnostic test for swines, makes sense. Immunochromatographic test should be adapted for this purpose. To design it we started a bio-guided identification of new proteins in cysticercus fluid. Proteins were analyzed using ion exchange chromatography and 2D separation and were selected by Western blot analysis using sera from infected/non infected pigs. Spots from the Coomassie-stained gel corresponding to these proteins were then analyzed by mass spectroscopy and proteins were identified using a bank of Expressed Sequence Tags (EST) of T. solium. Eighteen new proteins of interest were identified and nine were selected for further development.

Spatio-temporal prevalence of porcine cysticercosis in Madagascar based on meat inspection.

BACKGROUND: Taenia solium cysticercosis is a parasitic meat-borne disease that is highly prevalent in pigs and humans in Africa, but the burden is vastly underestimated due to the lack of official control along the pork commodity chain, which hampers long-term control policies. METHODS: The apparent and corrected prevalences of T. solium cysticercosis were investigated in pork carcasses slaughtered and retailed in Antananarivo (Madagascar), thanks to a 12-month monitoring plan in two urban abattoirs. RESULTS: Overall apparent prevalence was estimated at 4.6% [4.2 - 5.0%]. The corrected overall prevalence defined as the estimated prevalence after accounting for the sensitivity of meat inspection was 21.03% [19.18- 22.87%]. Significant differences among geoclimatic regions were observed only for indigenous pigs, with an apparent prevalence estimated at 7.9% [6.0 - 9.9%] in the northern and western regions, 7.3% [6.0 - 8.6%] in the central region, and 6.2% [4.7 - 7.8%] in the southern region. In the central region, where both exotic and indigenous pigs were surveyed, indigenous pigs were 8.5 times [6.7 - 10.7] more likely to be infected than exotic improved pigs. Urban consumers were more likely to encounter cysticercosis in pork in the rainy season, which is a major at risk period, in particular in December. Differences between abattoirs were also identified. CONCLUSION: Our results underline the need for improved surveillance and control programmes to limit T. solium cysticercosis in carcasses by introducing a risk-based meat inspection procedure that accounts for the origin and breed of the pigs, and the season.

Control of cysticercosis in Madagascar: beware of the pitfalls.

Taenia solium cysticercosis is a zoonosis of public health importance in areas where the disease is endemic, with significant economic impacts on human health and the swine industry. Several gaps remain in the epidemiology of the parasite and the strategies of control in developing countries. We detail the key factors to consider in Madagascar in terms of the porcine husbandry system, Taenia transmission cycle, and diagnosis of cysticercosis in pigs, in order to better estimate the sanitary and economic impacts of this parasitic disease as well as to define an integrated control program.

High prevalence of hepatitis E in humans and pigs and evidence of genotype-3 virus in swine, Madagascar.

Hepatitis E virus (HEV) causes an orofecal disease transmitted through poor hygiene environments, contaminated food (mainly pork products), or by contacts with infected animals. Very little data are currently available regarding the disease in the Southwestern Indian Ocean Islands. We report the first sero- and viro-survey for HEV in human and swine in Madagascar. A seroprevalence rate of 14.1% (60 of 427) was measured in slaughterhouse workers. Seroprevalence to HEV in pigs was estimated to 71.2% (178 of 250), strongly suggesting the existence of a zoonotic cycle. Three out of 250 pig livers (1.2%) tested HEV RNA-positive by quantitative polymerase chain reaction. Phylogenetic analyses based on 1-kb sequences of the ORF 2-3 identified these viruses as HEV genotype 3. Sequences clustered in a distinct Malagasy sub-clade, possibly representative of a new sub-genotype, for which the date of emergence was estimated around 1989. Further studies are needed to confirm other transmission routes of HEV to humans, especially through non-zoonotic cycles.