Evaluation of integrated control of three dog transmitted zoonoses: Rabies, visceral leishmaniasis and cystic echinococcosis, in Morocco.

The present study is a component of a major European research project, ICONZ (Integrated Control of Neglected Zoonoses). The objective of this component was to implement, and then evaluate an Integrated Control Intervention (ICI) against three dog transmitted zoonoses, namely rabies, visceral leishmaniasis, and cystic echinococcosis. This was undertaken in Sidi Kacem Province (northwestern Morocco) where 22 control and 22 treated douars (villages) were randomly allocated to two control and one evaluation interventions over a period of 13 months. Across the 44 douars, an overall total of 6922 dogs were registered, 4519 were vaccinated against rabies, and 2885 persons attended the complementary health education campaigns; whereas, within the 22 treated douars, 466 dogs received anti-sandfly collars (deltamethrin-impregnated) and 2487 were dewormed (praziquantel). Evaluation of the ICI, revealed that (i) a canine rabies vaccination coverage of 65.0% was achieved, (ii) the use of deltamethrin-impregnated collars for dogs provided highly significant protection (p = 0.01) against leishmaniasis infection of more than 44%, and up to 100% for dogs that had kept the collar until the end of the intervention, (iii) despite a non-significant difference (p>0.05), dog-deworming with praziquantel yielded a reduction in the rate of Echinococcus granulosus infection, and (iv) health education was successful in improving respondents' knowledge; However, the target communities remained unconvinced of the necessity of changing some of their risky behaviours. Lastly, the estimated total cost of the global intervention, including its research components, was US$ 143,050, of which 67% was for disease control work with clear evidence of significant economies of scale due to targeting three diseases together. It appears from this study that the integrated control approach against the three zoonoses was effective on both economic and logistical levels.

Use of FTA® card methodology for sampling and molecular characterization of Echinococcus granulosus sensu lato in Africa.

Cystic Echinococcosis is a parasitic disease caused by the cestode Echinococcus granulosus widely distributed in Africa. Monitoring of this parasite requires access to cyst samples on intermediate hosts observed at the slaughterhouse. In order to facilitate sampling in the field and analysis, the French National Reference Laboratory for Echinococcus spp. has developed a tissue derived from DNA sampling with FTA® card technology. The DNA samples were taken by applying the FTA® paper on the germinal layer after opening the cysts. The sampling technique was validated using frozen cysts (n = 76) stored in the laboratory and from field samples (n = 134) taken at the slaughterhouse by veterinarian technicians during meat inspection in Morocco, Mali and Mauritania. DNA was extracted after several weeks of storage at room temperature. PCR assays were performed using primers for generic cestode (cox1) and amplified fragments were sequenced. All samples taken in the lab and 80% of field samples were capable of molecular characterization. Cyst-derived DNA from FTA® samples can be useful for easy sampling, storage and rapid, safe and cheap shipment. The use of the FTA methodology will facilitate studies in the field to investigate the presence and genetic characterization of E. granulosus sensu lato in African countries.

Knowledge, attitudes and practices with regard to the presence, transmission, impact, and control of cystic echinococcosis in Sidi Kacem Province, Morocco.

BACKGROUND: This study is a component of a large research project on five major neglected zoonotic diseases (NZDs) including cystic echinococcosis and was undertaken in the Province of Sidi Kacem over a period of four years (April 2009-March 2013). METHODS: Questionnaires were administered at community level in a total of 27 communes and visits were made to all of the 10 abattoirs situated in the Province, to collect qualitative data on determinants of transmission for disease in humans and animals. More specifically, community knowledge, attitudes and practices related to cystic echinococcosis were assessed, as well as the extent to which local customs and behaviours may promote transmission. Abattoir infrastructure and practices, and their role in perpetuating disease transmission were also critically evaluated. RESULTS: The results show that only 50 % of people have heard of the disease, and of those, only 21 % are aware of the dog's role in disease transmission. Sixty-seven per cent of respondents stated that dogs are fed ruminant organs deemed unfit for human consumption. Owned dogs have access to the family home, including the kitchen, in 39 % of households. The extent of this close proximity between humans and animals is even more pertinent when one considers that dogs are omnipresent in the community, with an average of 1.8 dogs owned per household. The unrestricted access of dogs to abattoirs is a huge issue, which further promotes disease transmission. CONCLUSION: This study would suggest that the high prevalence of cystic echinococcosis in humans and animals in Morocco is largely due to three factors: 1) abundance of dogs 2) engagement in risky behaviour of the local population and 3) poor abattoir infrastructure and practices. This has serious implications in terms of the socio-economic impact of the disease, especially for rural poor communities.

Drug regimens for visceral leishmaniasis in Mediterranean countries.

Until the early 1990s, pentavalent antimony was the only documented first-line drug employed for the treatment of zoonotic visceral leishmaniasis (VL) in the Mediterranean, with reported cure rates exceeding 95% in immunocompetent patients. The emergence of antimony resistance in other endemic settings and the increase in drug options have stimulated re-evaluation of the current therapeutic approaches and outcomes in Mediterranean countries. A scientific consortium ('LeishMed' network) collected updated information from collaborating clinical health centres of 11 endemic countries of Southern Europe, Northern Africa and the Middle East. In contrast with the previous situation, VL is now treated differently in the region, basically through three approaches: (1) In Northern Africa and in part of the Middle East, pentavalent antimony is still the mainstay for therapy, with no alternative drug options for treating relapses; (2) In some European countries and Israel, both pentavalent antimony and lipid-associated amphotericin B (AmB) formulations are used as first-line drugs, although in different patients' categories; (3) In other countries of Europe, mainly liposomal AmB is employed. Importantly, cure rates exhibited by different drugs, including antimonials in areas where they are still in routine use, are similarly high (>/=95%) in immunocompetent patients. Our findings show that antimony resistance is not an emerging problem in the Mediterranean. A country's wealth affects the treatment choice, which represents a balance between drug efficacy, toxicity and cost, and costs associated with patient's care.

Modelling the transmission of Echinococcus granulosus in dogs in the northwest and in the southwest of Morocco.

Echinococcus granulosus (E. granulosus) infection was studied in 151 dogs in two regions of Morocco: 68 dogs in the northwest (Loukkos) and 83 dogs in the southwest (Tiznit). The mean prevalence rates of echinococcosis in dogs were 58.82% (46.23-70.63%) in Loukkos and 55.42% (44.10-66.34%) in Tiznit and the mean abundances of E. granulosus per dog were 75 (59-93) and 547 (504-595), respectively. The mean abundance of E. granulosus in dogs was fitted to a negative binomial distribution by the maximum likelihood techniques to define parameters. E. granulosus was aggregated in dogs in the two regions. The prevalence of infection and the abundance of E. granulosus in dogs were fitted to mathematical models in order to determine if the parasite population is partly regulated by definitive host immunity. The best fit was obtained with the models assuming the presence of immunity. The mean time of exposure to infection was similar in the two regions and ranged from 8 months to about 2 years. The infection pressures (number of E. granulosus) obtained per dog each year were 65 (8-294) in Loukkos and 476 (316-886) in Tiznit. The proportion of dogs susceptible to infection was still high along the life of the dogs in Loukkos, while it was not different from zero in old dogs of Tiznit.

Epidemiological study of the cystic echinococcosis in Morocco.

The objectives of this epidemiological study on cystic echinococcosis (CE) in Morocco (2001-2004) were to update the prevalence of CE in different animal species living in the most important areas of the country and to collect protoscoleces and germinal layers for genetic research purposes. The post mortem inspection concerned 2948 sheep, 2337 goats, 618 cattle, 482 camels and 455 equines (325 horses, 60 mules and 70 donkeys) in five different regions: the Rif (Mediterranean coast and high mountains of the Rif), the Loukkos (Atlantic northwest plain), the center (Rabat and Casablanca regions), the Middle Atlas mountains and the south (arid and semi desert areas). The global CE infection prevalence rates obtained were 22.98% in cattle, 10.58% in sheep, 12.03% in camels, 17.80% in equines and 1.88% in goats. The infection rates were especially high in the Middle Atlas in cattle (48.72%) and in the Loukkos in cattle and sheep (37.61 and 31.65%, respectively). The majority of infected cattle (49.6%) and sheep (52.1%) had hydatid cysts in both liver and lungs. Except for cattle, the liver was more infected than lungs in all the other animal species. Animals more than 5 years old were the most infected in all species. The mean CE infection rates of these animals were about 56% in cattle, 40% in sheep, 20% in camels, 17.80% in equines and 7% in goats. These rates were much higher in the Loukkos (85% of cattle and 59% of sheep) and in the Middle Atlas (68% of cattle and 45% of sheep) than in the other regions. Results showed that Echinococcus granulosus is in an endemic steady state with no evidence of protective immunity in the intermediate hosts. The mean numbers of infections per year are 0.099 for cattle, 0.063 for sheep, 0.03 for camels and 0.010 for goats.

Signature of an early genetic bottleneck in a population of Moroccan sardines (Sardina pilchardus).

Fishery assessment models meant to determine sustainability of commercial marine fish failed to predict recent stock collapses due to overexploitation. One flaw of assessment models is that they strongly rely on catch and age-composition statistics, but largely ignore the genetic background of the studied populations. We examined population genetic structure of sardine (Sardina pilchardus) in the centraleastern and northeastern Atlantic Ocean and Mediterranean Sea to aid fishery management of this heavily fished small pelagic species. We found that sardine has a striking mitochondrial control region, and sequenced a fragment of 387 bp of its 5'-end in 261 individuals collected off the coasts of Morocco (Dakhla, Tantan, Safi, Larache, and Nador), Portugal (Quarteira), Spain (Pasajes, Barcelona), and Greece (Kavala). High levels of haplotypic diversity rendered a rather unresolved NJ phylogeny. The recovered tree had no phylogeographic structuring except for the clustering of 13 individuals of Safi. In contrast, individuals grouped together according to the presence or absence of a 13-bp insertion in the sequence. Phi(ST) pairwise comparisons and molecular variance analyses supported genetic differentiation between the population of Pasajes (Bay of Biscay), and those of the Mediterranean Sea and Moroccan coast, with a contact zone around the Strait of Gibraltar. This result confirms the existence of two subspecies, S. pilchardus pilchardus and S. pilchardus sardina that were previously identified based on meristics and morphometry. Mismatch distribution analysis showed that sardine populations are expanding since the Pleistocene. Surprisingly, the population of Safi showed strong and statistically significant levels of genetic differentiation that could be related with isolation and genetic drift. Comparative analysis of the Safi population versus the rest including mismatch distributions, and a Bayesian skyline plot suggest that the Safi population likely underwent an early genetic bottleneck. The genetic singularity of the Safi population could have been responsible for the historical collapse of this sardine stock in the 1970s.