Echinococcus granulosus infection in dogs in Sidi Kacem Province (North-West Morocco).

This study was undertaken in the Province of Sidi Kacem in northwest Morocco between April 2010 and March 2011. The main objective of the study was to determine the prevalence of Echinococcus granulosus (Eg) infection in owned dogs. This province was selected as a case study because of the social conditions, geographic and climatic diversity making it a model representative of many parts of Morocco. The survey was carried out in 23 rural communes and in the 5 municipalities (urban districts) of the Province and sampling was undertaken in randomly selected households. A total of 273 owned dogs comprising 232 from the 23 rural communes (rural dogs) and 41 from the 5 municipalities (urban dogs) were tested. Arecoline hydrobromide purgation was selected as the diagnostic method of choice to enable visualisation of expelled worms by dog owners, thereby imparting messages on the transmission mode of Eg to humans and farm animals. Of the 273 dogs tested, purgation was effective in a total of 224 dogs (82.1%). The overall estimated prevalence of Eg infection was 35.3% (79/224, 95% CI 22.3-47.0%). Dogs inhabiting rural communes were at greater risk of infection (38.0%, 95% CI 31.1-45.3%) than dogs roaming in municipalities or urban areas (18.8%, 95% CI 7.2-36.4%) and the prevalence of infection was higher in those inhabiting rural communes with slaughterhouses (62.7%, 95% CI 48.1-75.9%) than in communes without (29.1%, 95% CI 21.7-37.2%). This first assessment of Eg infection in Sidi Kacem Province indicates a key role of rural slaughterhouses in parasite transmission to dogs.

Integrated health messaging for multiple neglected zoonoses: Approaches, challenges and opportunities in Morocco.

Integrating the control of multiple neglected zoonoses at the community-level holds great potential, but critical data is missing to inform the design and implementation of different interventions. In this paper we present an evaluation of an integrated health messaging intervention, using powerpoint presentations, for five bacterial (brucellosis and bovine tuberculosis) and dog-associated (rabies, cystic echinococcosis and leishmaniasis) zoonotic diseases in Sidi Kacem Province, northwest Morocco. Conducted by veterinary and epidemiology students between 2013 and 2014, this followed a process-based approach that encouraged sequential adaptation of images, key messages, and delivery strategies using auto-evaluation and end-user feedback. We describe the challenges and opportunities of this approach, reflecting on who was targeted, how education was conducted, and what tools and approaches were used. Our results showed that: (1) replacing words with local pictures and using "hands-on" activities improved receptivity; (2) information "overload" easily occurred when disease transmission pathways did not overlap; (3) access and receptivity at schools was greater than at the community-level; and (4) piggy-backing on high-priority diseases like rabies offered an important avenue to increase knowledge of other zoonoses. We conclude by discussing the merits of incorporating our validated education approach into the school curriculum in order to influence long-term behaviour change.

Echinococcosis/hydatidosis: a severe threat in Mediterranean countries.

Echinococcosis/hydatidosis is one of the most important parasitic zoonotic diseases in the world. Both cystic hydatidosis (CE) caused by Echinococcus granulosus and alveolar echinococcosis (AE) caused by E. multilocularis have been reported in several countries of the Mediterranean region (MR). E. granulosus has always been present in the MR and is the most common species. This parasite depends on the dog-sheep cycle and is actively transmitted in all pastoral regions where sheep, cattle and camelids predominate. E. multilocularis occurs only sporadically in limited areas of France, Serbia and Montenegro, Turkey, Tunisia and Morocco. However recent evidence indicates that it is spreading into other regions of the Mediterranean. Due to the lack of well-documented data, and to the fact that CE is not a notifiable disease in the majority of M countries, the precise incidence and prevalence of CE in humans and animals are not known. Published data suggests that prevalence is rather high in North Africa, Turkey, Greece, and in several regions of Italy and Spain. CE is an increasing public health and socio-economic concern due to the considerable morbidity rates that give rise to high economic losses both in the public health sector and in the livestock industry. Hospitalisation for human CE lasts from 2 weeks to more than 1 month in case of surgery. A number of factors contribute to the increase of prevalence and to the spreading of CE in the MR. These include the diversity of livestock production systems (predominantly extensive, traditional animal husbandry), small, ill-equipped and unsupervised slaughter-houses, illegal and family slaughtering, low public awareness of hydatid diseases, and the high population of stray dogs. Cyprus is the only country where an eradication programme has been successfully implemented. There have been, however, important developments in the last decade in CE epidemiology, in the diagnosis of canine infection, in strain characterisation and in immune strategies against CE in animals. This scientific progress, together with effective health education programmes, will likely improve control programmes and reduce the time required to achieve significant decreases in prevalence or eradication.

First detection and genetic typing of Leishmania infantum MON-24 in a dog from the Moroccan Mediterranean coast: genetic diversity of MON-24.

As in the countries edging the Mediterranean basin, Leishmania infantum zymodeme MON-1 is the main causative agent of visceral leishmaniasis in Morocco, where visceral leishmaniasis is most active in the North-Eastern slopes of the Rif mountains. The dog was confirmed to be the main reservoir of L. infantum MON-1, while the reservoir of L. infantum MON-24 causative agent of both infantile visceral leishmaniasis and cutaneous leishmaniasis has not yet been identified. Here we report the first detection of this last zymodeme in a dog in Morocco. The isolated strain was first identified by the use of genotyping markers and confirmed by isoenzyme analysis. Phylogenetic analysis with the use of concatenated sequences from 26 Leishmania donovani complex strains revealed strong geographical correlation with the MON-24 strain from Morocco clustering with other East African strains whereas two other MON-24 strains clustered with L. infantum strains. Interestingly, the two distinct populations of MON-24 identified with the use of genotyping markers cannot be distinguished by multilocus enzyme electrophoresis.

Identification of two highly performing Leishmania infantum antigens for serodiagnosis of canine leishmaniosis.

In the aim of improving serodiagnosis of canine leishmaniosis, we analysed the humoral immune response of dog against Leishmania infantum parasite. The antigenic reaction of L. infantum polypeptides with sera from 31 dogs with parasitologically confirmed leishmaniosis was studied by using the immunoblot technique. Electrophoretic profile of the parasite extract showed more than 50 polypeptides, with molecular weights ranging from 12 to 170 kDa. Among these polypeptides, 37 antigen components, ranging from 14 to 91 kDa, were recognised by antibodies of L. infantum infected dogs. Three polypeptides (14, 16 and 76 kDa) reacted with all of the 31 serum samples. The other most frequently recognised antigens were those of 29.5, 32, 46, 59 and 66 kDa with a sensitivity of 87.1%, 93.6%, 96.8%, 87.1% and 80.6%, respectively. The 14 and 16 kDa bands were the most intense and remained detectable until a serum dilution of 1:6400. No reaction of these two major antigens was observed with sera collected from 50 Leishmania-free dogs, living in the leishmaniosis-free region of Rabat in Morocco, whereas the crude antigen used in IFAT or ELISA lead to three false positive results. Four antigen components of 29, 41, 55, and 70 kDa were recognised by some sera samples from negative controls. These results demonstrated the potential interest of the fractions of 14 and 16 kDa in immunodiagnosis of canine leishmaniosis.

[Wound myiasis due to Wohlfahrtia magnifica. First human case in Morocco]. / Myiase des plaies dues à Wohlfahrtia magnifica (Schiner, 1862). Premier cas humain au Maroc.

Wohlfahrtia magnifica is a frequent fly in countries with high breeding activity: Australia, South Africa or Arab country The parasitological infestation occurred essentially during the summer, favourable period to the biological evolution of the flies. In this work, we report one case of wound myiasis that complicates a cutaneous lesion caused by larvae of Wohlfahrtia magnifica. This observation constitutes the first human case in our country.

Camel trypanosomosis in Morocco. 2. A pilot disease control trial.

A first epidemiological study on camel trypanosomosis was conducted in 1997 and 1998 in five Moroccan Saharan provinces. It revealed two major foci at Merzouga (Tafilalet province) and Zagora (Ouarzazate province) where tour camel seroprevalences were 35.4 and 43.3%, respectively, for each region. The first area was chosen in the present study as a pilot region for a control program based on epidemiological data with the aim of evaluating the efficiency of the trial in terms of seroconversion, incidence and health condition. After finding the endemic focus at Merzouga, four other field studies were organised in this area, at 4-month intervals, in order, firstly, to control the camel reservoir by treating all the camels found in the region during the visit using melarsomine (Cymelarsan, Rhône-Mérieux, the only trypanocide registered in Morocco), and, secondly, to collect blood samples for studying temporal changes in seroprevalence as well as in anaemia rate. All the 291 tour camels of the area, that were present in at least one of the field visits were examined and included in the trial. Among them, 162 were treated once, 84 twice and 41 three times. A total of 676 serum samples were collected from these camels. At the end of the trial--1 year later--seroprevalence had dropped from 58% (May 1998) to 19% and the mean Ab-ELISA value had decreased from 0.52 (before treatment) to 0.12 (which is below the threshold level), after the third campaign. The haematocrit of the animals was also slightly improved, indicating that trypanosomosis was a major cause, but it was not the only incriminated factor. A seasonal effect was observed. The overall seroprevalence was reduced between spring 1999 and summer 1999, without treatment, whereas a negative effect was recorded on packed cell volume, more camels being found anaemic in summer than in spring. The application of these strategic treatments was associated with a large reduction in the seroprevalence of trypanosomosis, but there was no complete eradication because of the continual exchange of animals from other foci such that of Zagora. The overall seroconversion after 1 year reached 55% and no post-treatment complication was recorded, whereas the incidence due to new infections approached 4.3%. Moreover, several factors like the unusual way of life of the dromedaries, including stress, excessive work, overcrowding and continual contact with other animals coming from other infected regions contribute to the persistence of the disease.

A prospective, randomized, controlled trial comparing highly purified hMG with recombinant FSH in women undergoing ICSI: ovarian response and clinical outcomes.

BACKGROUND: To assess the clinical profile and efficacy in assisted reproductive treatment of a new human-derived highly purified (HP) menotropin, we compared HP hMG and recombinant (r) FSHalpha use in ICSI within a prospective, randomized, controlled study. METHODS: 100 infertile women were treated with HP hMG (50 patients) or rFSHalpha (50 patients). All patients received the same daily gonadotrophin dose (150 IU) following GnRH agonist suppression (long regimen) until more than three follicles >17 mm and estradiol (E(2)) levels >600 pg/ml were reached. Patients were monitored with daily LH, FSH, hCG, estradiol (E(2)), progesterone, and testosterone measurements; and alternate day pelvic ultrasound. RESULTS: Treatment duration (11.1 +/- 0.4 versus 12.9 +/- 0.5 days, P < 0.05) and gonadotrophin dose (22.4 +/- 1.0 versus 27.0 +/- 1.5 ampoules, P < 0.05) were lower in the HP hMG group. Conversely, peak pre-ovulatory E(2) (1342 +/- 127 versus 933 +/- 109 pg/ml, P < 0.005); and area under the curve of E(2) (3491 +/- 350 versus 2602 +/- 349 pg/ml.day, P < 0.05), immunoreactive serum FSH (65.9 +/- 2.1 versus 48.8 +/- 1.8 IU/l.day, P < 0.001). and hCG (1.7 +/- 0.3 versus 0.0 +/- 0.0 IU/l/day, P < 0.001) during treatment were higher in the HP hMG group. Cycle cancellation rates, transferred embryo number, pregnancy rates per started cycle (30 versus 28%) and per embryo transfer (35 versus 35%) and miscarriage rates (6 versus 6%) were not significantly different. CONCLUSIONS: HP hMG treatment was associated with: (i) a more efficient patient response, as reflected by reduced treatment duration and gonadotrophin requirements; (ii) increased serum levels of hCG, E(2), and immunoreactive FSH during treatment; (iii) an ICSI outcome indistinguishable from rFSHalpha.

[Canine leishmaniasis in the Rif mountains (Moroccan Mediterranean coast): a seroepidemiological survey]. / Leishmaniose canine dans le Rif (côte méditerranéenne marocaine): enquête séro-épidémiologique.

A sero-epidemiological survey has been conducted in several localities of the province of Nador to investigate canine leishmaniasis in the North-Eastern slope of the Rif mountains (Mediterranean coast of Morocco). Serum samples collected from 257 dogs were analysed using indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA) to detect anti-Leishmania infantum antibodies. Forty eight (18.7%) of the screened dogs were IFAT positive and 54 (21.0%) were ELISA positive; the concordance of the two methods was 96.1%. The prevalence of infection is significantly higher in dogs more than four years of age whereas no significant difference in prevalence of infection was seen between males and females. The frequent symptoms observed in seropositive dogs were the enlargement of lymph nodes (57.4%), emaciation (51.9%) and skin involvement (25.9%). However, 38.9% of those dogs showed no one of the major symptoms of visceral leishmaniasis. Leishmania isolated from three of the examined dogs was identified as L. infantum MON-1. These results show that the North-Eastern slope of the Rif mountains is one of the most active Mediterranean areas of visceral leishmaniasis and confirm that the dog is the main reservoir of L. infantum.

Camel trypanosomosis in Morocco 1: results of a first epidemiological survey.

An epidemiological survey of camel trypanosomosis was conducted for the first time in Morocco in 1997 and 1998. Five provinces located in the south and southeast of the High Atlas mountain chain were included in this study. A total number of 1460 serum samples were collected and tested by a card agglutination test for trypanosomosis (CATT) and enzyme-linked immunosorbant assay (Ab-ELISA) to detect anti-Trypanosoma evansi antibodies. The overall seroprevalence was 14.1% by CATT and 18.2% by Ab-ELISA. Two very active foci were identified in Zagora and Merzouga with high Ab-ELISA prevalences of 43.3 and 35.4%, respectively, for each area. The concordance between the CATT and Ab-ELISA was 94.1%. The age-related distribution of seroprevalence showed a tendency for the infection rate to increase with age up to a maximum in the 7-10 year-old group. Clinical examination revealed that enlargement of lymph nodes was the most frequent sign in seropositive animals (79.7%). The comparison of haematocrit values of seropositive and seronegative animals showed a significant difference that indicated severe anaemia in infected animals. Isolation of the parasite by inoculation of mice allowed us to collect 15 T. evansi isolates from Zagora and three from Merzouga. The study showed that the Saharan provinces of Morocco were slightly affected by camel trypanosomosis but two hyperendemic foci, Zagora and Merzouga, were found to be sources of contamination for the surrounding regions.

Triplet pregnancy and delivery after intracytoplasmic injection of round-headed spermatozoa.

Intracytoplasmic sperm injection (ICSI) of round-headed spermatozoa into mature oocyte resulted in normal fertilization, embryo development and pregnancy in a 28 year old female. The husband had a long history of primary infertility. Three ICSI attempts were carried out and fertilization and embryo development occurred in all trials. However, only the third trial led to a pregnancy, which proved to be quadruplet after the transfer of four embryos. One embryo vanished and the remaining triplets were delivered at 35 weeks of gestation by Caesarean section. Two of the babies, a boy weighing 2000 g and a girl weighing 2250 g at birth were discharged in a good condition 1 week after delivery and the third baby, a boy weighing 1550 g, was discharged 3 weeks after delivery.

Protective oral immunization of chickens against Eimeria tenella with sporozoite surface antigens.

Antigens were extracted from the surface of Eimeria tenella sporozoites with a solution containing Triton X 100 (1%), sodium dodecyl sulphate (0.5%) Na deoxycholate (1%) and EDTA (1 mM). After removal of the detergents, these surface antigen preparations conferred an immunity that protected chickens against a subsequent infection (10(4) sporulated oocysts). The best results were obtained after two 250 micrograms injections of Al(OH)3 adsorbed antigens (oocyst output per g caecal material on Day 7 post infection: 2.39 x 10(7) +/- 0.32 x 10(7) oocysts for controls and 7.37 +/- 10(6) +/- 3.19 x 10(6) oocysts for vaccinated birds) and after four gastric intubations of liposome entrapped antigens (oocysts output on Day 7 postinfection: 2.75 x 10(6) +/- 2.02 x 10(6) g-1 caecal material). These results represented respectively 70 and 88% protection indexes. Studies on the systemic and local antibody response after one or several infections of chickens with the parasite indicated at least 20 different molecules in the detergent antigens which are classified after immunoblotting according to their properties.

Effect of temperature and relative humidity on survival of eggs and infective larvae of Ostertagia circumcincta.

Survival of Ostertagia circumcincta eggs in faeces kept at 4 degrees C and of third stage larvae (L3) kept in distilled water at -50, -25, -10, 4, 16, 25 and 35 degrees C was studied. The effect of relative humidity (RH) of 95, 75, 50 and 30% at 16, 25 and 35 degrees C on the survival of L3 was also studied. The survival of eggs at 4 degrees C was high during the first week, followed by a gradual decline to 10% by Day 22 of storage. The L3 survived for 7 weeks at -50 degrees C, 9 weeks at -25 degrees C, 13 weeks at -10 degrees C, and 7 weeks at 35 degrees C. By 16 weeks, at the termination of the experiments, 74%, 76% and 23% of L3 were still alive at 4 degrees C, 16 degrees C and 25 degrees C, respectively. At all temperatures tested, the L3 survived longer at RH of 30 and 50% than at 75 and 95%. At 35 degrees C, L3 survived up to 6 weeks at 30%, 5 weeks at 50% and 1 week at 75 and 95% RH. At 16 and 25 degrees C and RH of 30 and 50%, over 70% of L3 were still alive at the end of experiments at the seventh week. At 25 degrees C, 40% of L3 survived for up to 5 weeks at 75% RH and 5% survived for 6 weeks at 95% RH.(ABSTRACT TRUNCATED AT 250 WORDS)

Seasonal dynamics of Fasciola hepatica burdens in grazing Timahdit sheep in Morocco.

Seasonal transmission of Fasciola hepatica was observed in sentinel sheep and the dynamics of the snail intermediate host, Lymnaea truncatula, was followed over a 3-year study period in the Middle-Atlas mountains in Morocco. High fluke burdens were recorded in both lambs and ewes in the fall and winter, suggesting that transmission occurred in late spring. Fluke burdens ranged from one to 302 in ewes and from one to 345 in lambs. Infections with 200 or more flukes were always fatal. A unique feature of this study was the annual cyclical fluctuation of the fluke burdens. Burdens reached maximum levels during the winter and then declined to low numbers by late spring and summer. This suggested self-regulation which may be dependent on breed resistance or may be related to forage factors, including lack of forage (nutritional stress). Snail populations were cyclical and correlated with fluke transmission as observed in the sentinel sheep. The weather was observed to affect the snail populations which in turn limited fluke transmission.

[Gastrointestinal strongylosis and malabsorption of nutrients]. / Strongyloses gastro-intestinales et malabsorption des nutriments.

The role of malabsorption of nutrients in the mechanism of reduction in productions of the ruminants infested with gastrointestinal nematodes has been a controversial question because of the compensatory absorption and reabsorption in distal part of the small intestine. However, during the last decade, leading research showed that malabsorption has a major role in this mechanism because of 1) compensatory absorption and reabsorption remain limited to some nutrients, 2) large quantities of incompletely digested nutrients exceeding the intestine capacity, and 3) poor reutilization of reabsorbed nutrients.

Effect of temperature on development of the free-living stages of Ostertagia circumcincta.

Hatching of the eggs of Ostertagia circumcincta was studied by recovering them from faeces and incubating them in distilled water at temperatures of 4, 16, 25 and 35 degrees C. Hatching occurred at all the temperatures. The rate of hatching increased with the rise in temperature. Development of larvae to the infective third stage (L3) was studied in faecal cultures incubated at 4, 16, 25, 30, 35, 40 and 45 degrees C. Except at 4 degrees C, L3 developed at all temperatures, the optimum temperature being 16 degrees C. The rate of development of L3 increased with the rise in temperature. This resulted in a corresponding decrease in the percentage recovery of larvae.

Parasites of stray dogs in the Rabat region, Morocco.

Eighteen parasites were identified from 57 stray dogs from the urban and rural areas of Rabat. The prevalence of the species of parasites was: Ctenocephalides (C. canis, C. felis) 94.7%, Rhipicephalus sanguineus 68.5%, Linguatula serrata, Babesia canis, Isospora canis each 3.5%, Toxocara canis 7%, Dirofilaria immitis, Taenia hydatigena, T. pisiformis each 12.3%, Dipetalonema dracunculoides 10.5%, Ancylostoma caninum 17.5% Echinococcus granulosus 22.8%, Toxascaris leonina 33.3%, Dipylidium caninum 40.4%, Spirocerca lupi 54.5%, Unicinaria stenocephala 79%. Multiple infections (two to nine parasites) were seen in 56 dogs. T. canis was present only in dogs younger than one year of age. D. immitis, D. dracunculoides, T. hydatigena and T. pisiformis occurred only in animals older than one year. A. caninum, U. stenocephala, S. lupi, D. immitis were more common in males than females. E. granulosus was never detected in urban dogs.