Calicophoron daubneyi and Fasciola hepatica: characteristics of natural and experimental co-infections of these digeneans in the snail Lymnaea glabra.

A retrospective study on different Lymnaea glabra samples collected from central France between 1993 and 2010 was carried out to determine the prevalence of natural co-infections with Calicophoron daubneyi and Fasciola hepatica, and to specify the composition of redial burdens. Experimental infections of L. glabra performed during the same period of time were also analysed to study metacercarial production of each digenean in co-infected snails. Controls were naturally or experimentally co-infected Galba truncatula. In natural co-infections, prevalence was 0.7% in L. glabra (186/25,128) and 0.4% in G. truncatula (137/31,345). Low redial burdens were found in these snails, with F. hepatica rediae significantly more numerous in L. glabra than in G. truncatula (7.5 per snail instead of 5.2). In contrast, the total numbers of C. daubneyi rediae in both lymnaeids were close to each other (4.3 and 3.0 rediae, respectively). In experimentally co-infected groups, prevalence was greater in G. truncatula than in the other lymnaeid (6.3% instead of 3.0%). Significantly shorter patent periods and lower metacercarial production for each digenean were noted in L. glabra than in G. truncatula. However, in both lymnaeids, the two types of cercariae were released during the same shedding waves and several peaks during the patent period were synchronous. In spite of a greater shell height for L. glabra, metacercarial production of both digeneans in co-infected snails was lower than that in G. truncatula, thus indicating a still incomplete adaptation between these French L. glabra and both parasites.

Prevalence of Fasciola hepatica in Galba truncatula detected by Multiplex PCR in the province of El Tarf (Algeria).

The prevalence of Fasciola hepatica infection in the snail Galba truncatula was studied for the first time in Algeria using Multiplex PCR. A total of 722 individuals were collected from 11 typical habitats (temporary ponds, road ditches and puddles) distributed in five localities of the El Tarf province, known for endemicity of fascioliasis. Snails were divided in 75 groups and DNA extraction was performed using Chelex® (Biorad, Nazareth Eke). Two primers were used, the first is specific for 124 bp DNA fragment of Fasciola sp and the second one represents the ITS-2 lymnaeids sequence (500-600 bp). The prevalence of the infection was estimated to be 46.66% for pools (35/75 were positive). The bursting out of the pools show that 75 snails were carriers of F. hepatica which gives an overall infection rate of about 10.74%. The results of snails infection according to their sizes showed significant differences in the studied size classes, thus snails of big sizes were the most infected. Based on these data, epidemiological implication of Multiplex PCR as a fully reliable technique to highlight high risks periods of fasciolosis will be of a great interest.

Natural light influences cercarial emergence of Calicophoron daubneyi but not that of Haplometra cylindracea from temperature-challenged Galba truncatula.

Laboratory investigations on Galba truncatula experimentally infected with Calicophoron daubneyi were carried out to study the influence of natural light and sky nebulosity on cercarial emergence in snails subjected every week outdoors to a thermal shock (a mean of 12 °C for 3 h) during the patent period. The same study was also performed in G. truncatula naturally infected with Haplometra cylindracea according to the same protocol. Compared to infected controls always reared indoors at 20 °C, the number of cercariae emerging from snails subjected outdoors to a thermal shock was significantly greater, whatever parasite species. Natural light had an effect on snails releasing C. daubneyi cercariae after the thermal shock because their numbers were significantly higher between 601 and 1,200 lx and for the highest nebulosity values (7-8 octas). In contrast, the type of light used during thermal shock did not influence cercarial emergence of H. cylindracea because the numbers of cercariae per shedding snail noted under natural light or a 3,000-lx artificial light did not significantly differ from each other. Most snails releasing H. cylindracea cercariae were significantly more numerous for light levels higher than 3,000 lx. As for Fasciola hepatica cercariae, natural light had a significant influence on cercarial emergence of C. daubneyi, and this behavioural particularity for cercariae of both digeneans might be due to the fact that both species develop in the same ruminants and the same snail host.

Paramphistomum daubneyi: characteristics of infection in three lymnaeid species.

Experimental infections of two South American lymnaeids (Lymnaea neotropica and L. viatrix var. ventricosa) with Paramphistomum daubneyi were carried out to determine if these snail species could sustain larval development of this digenean and, if so, to specify their potential for cercarial production. A French population of Galba truncatula infected and raised according to the same protocol served as controls. In both experiments, prevalence of P. daubneyi infections in snails did not significantly differ from each other. In snail groups evaluated for cercarial shedding (first experiment), a significantly lower number of shed cercariae was noted for L. neotropica, while those from G. truncatula and L. v. ventricosa did not differ significantly from each other. Dissection of infected snails at day 65 post-exposure at 20°C (second experiment) found significantly lower burdens of P. daubneyi rediae and cercariae in the bodies of L. neotropica than in those of G. truncatula and L. v. ventricosa. Compared to total cercarial production observed in dissected snails, the percentage of cercariae which exited from snails was 75.6% for G. truncatula, 21.6% for L. neotropica, and 91.4% for L. v. ventricosa. This last species seems to be a good candidate for metacercarial production of P. daubneyi.

Prevalence and intensity of Paramphistomum daubneyi infections in cattle from north-eastern Algeria.

Parasitological investigations on ruminal paramphistomosis were carried out in 2033 cattle over a 14-month period in three Algerian slaughterhouses to determine the prevalence and intensity of this disease. The prevalence of infection varied from different slaughterhouses (1.2% at El Khroub, 7.5% at Ferdjioua and 12.1% at Jijel). The worm burden was significantly higher in cattle from El Khroub than for those slaughtered in the other two sites (a mean of 984.1 parasites compared with 87.5 and 140.7 at Jijel and Ferdjioua, respectively). No significant correlation between prevalence, intensity of infection and age of cattle was noted. Overall, females were significantly more infected than males, whereas the sex of animals was independent of worm burdens recovered in slaughtered cattle. The climatic conditions of provinces in which the slaughterhouses were located (a more temperate climate for Jijel and semiarid conditions for the other two provinces) may partly explain variations noted in prevalence and intensity of infection. The present results will contribute to our understanding of the epidemiology of Paramphistomum daubneyi infections in north-eastern Algeria.

Total cercarial output in two populations of Galba truncatula experimentally infected with Fasciola hepatica.

Experimental infections of pre-adult Galba truncatula (two populations) using one, two, three or five miracidia of Fasciola hepatica per snail were carried out to determine the total number of cercariae produced by snails when they were sacrificed at day 70 post-exposure (at 24 degrees C). When the number of miracidia used for each snail increased at exposure, significant numerical augmentations of live rediae and of intraredial differentiating cercariae were noted. In contrast, only insignificant differences between mean numbers of free cercariae were found. The number of metacercariae produced by each snail (after shedding and/or after dissection) increased in infections with up to three miracidia per snail in each population and strongly decreased in infections with five miracidia per snail. Compared to the other types of infections, the delay of cercarial differentiation noted in the five-miracidia groups was probably due to the volume of the snail host, which would be too small to assure the complete development of five sporocysts.

The proportion of liver excised in Algerian abattoirs is not a good indicator of Fasciola hepatica infections in local cattle breeds.

Clinical and serological investigations on 175 cattle were carried out in 1999 at the slaughterhouse of Jijel, northeastern Algeria, to verify if partial ablation of a liver infected with Fasciola hepatica represents a good epidemiological indicator in the case of fasciolosis, as partial excision of the liver is widespread throughout North African countries. This study was also performed to determine if there is a direct relationship between the quantity of liver confiscated for fasciolosis and the serological response of slaughtered animals. A significant relationship between highly infected livers and positive serological titres was noted. However, two groups of results were conflicting: the presence of macroscopically healthy, undamaged livers with a positive serology (8.3% of 120 cattle), or the existence of highly damaged livers with a negative serology (13.0% of 31 cattle). These results indicate that partial removal of the liver in the case of fasciolosis is unreliable, so this method does not represent a good tool to establish epidemiological data on this disease.

The presence of uninfected Omphiscola glabra in a breeding of infected Galba truncatula enhanced the characteristics of snail infections with Fasciola hepatica.

Experimental infections of Galba truncatula with Fasciola hepatica were carried out under laboratory conditions to determine if the presence of another lymnaeid, Omphiscola glabra, during snail breeding had an indirect effect on the growth of G. truncatula and to analyse consequences on cercarial production. When the two lymnaeids are raised together, the survival of G. truncatula at day 30 post-exposure (PE), the prevalence of snail infections, and the shell height of cercariae-shedding snails at day 45 PE were significantly higher. By contrast, the other parameters characterizing snail infections only showed insignificant variations. The origin of O. glabra (living in a snail community, or monospecific population) used for the breeding of G. truncatula did not have a significant effect on the values of each parameter. Even if variations in the mean numbers of metacercariae were insignificant, the greater survival of G. truncatula at day 30 PE and the higher numbers of cercariae-shedding snails in the groups living with O. glabra allowed to obtain a higher total number of larvae than in alone-raised groups of G. truncatula.

Preliminary studies on the prevalences of natural fasciolosis in cattle, sheep, and the host snail (Galba truncatula) in north-eastern Algeria.

Three series of investigations on natural infections with Fasciola hepatica were carried out in north-eastern Algeria): (1) on different samples from cattle and sheep slaughtered at Constantine and Jijel from 1994 to 1996, (2) from 31 cattle- and sheep-breeding farms from 1999 to 2001, using serology on blood samples, and (3) on overwintering Galba truncatula from four populations, in 2002 and 2003. Significantly higher prevalences of fasciolosis were found in the cattle and sheep from Jijel (27.0% in slaughtered cattle and 27.3% in cattle from farms, compared to 9.1% and 6.3%, respectively, in cattle from Constantine). No significant differences in the prevalences were recorded between 1994 and 1996 and 1999 and 2001 for each species of ruminant, whatever the department considered. The infection rates for overwintering snails were also higher in the two populations from Jijel compared with those from the department of Constantine (a mean of 4.6-5.9% instead of 2.6-3.1%)). These results may be explained by the climatic conditions in the department of Jijel (an annual rainfall of 750-900 mm instead of 350 mm at Constantine), and the mode of anthelminthic treatment (the periods of treatment would be inappropriate in the case of Jijel).

Variability in the prevalence of infection and cercarial production in Galba truncatula raised on a high-quality diet.

Two experiments using seven populations of Galba truncatula were carried out to analyse the effect of food quality (cos lettuce only, or cos lettuce+Tetraphyll) on the characteristics of infections: (1) in a single population of G. truncatula infected by one of three digenea (first experiment), and (2) in seven populations of G. truncatula differing in their susceptibility to Fasciola hepatica miracidia (second experiment). In most groups, food quality did not have a significant effect on the survival of snails. The prevalence of infection in five populations was significantly higher in snails raised on lettuce+Tetraphyll (first and second experiments), whereas it was close to those noted in lettuce only-reared groups in the last two populations (second experiment). Despite the higher growth of cercariae-shedding snails when raised on the mixed diet, no significant differences were noted. Significant effects of parasite species (first experiment) and of snail population (second experiment) on the life-spans of cercariae-shedding snails were noted, whereas food quality did not influence this parameter. Except for a single snail population, cercarial production in groups raised on lettuce+Tetraphyll was significantly higher than that in groups on lettuce. The variability noted in the prevalence of snail infections and in the intensity of cercarial shedding might be explained by differences in the susceptibility of snail populations to F. hepatica infections, and/or by the fact that Tetraphyll would not have the same appetency for all populations of G. truncatula.

[Bovine hypodermosis in north-eastern Algeria: prevalence and intensity of infestation]. / Hypodermose bovine dans le Nord-Est algérien: prévalence et intensité d'infestation.

Between 1989 and 1997 different factors related to warble fly infestation (prevalence, intensity, climate, breed) were studied in north-eastern Algeria. Mean prevalence during the period was 76%. This figure was obtained from a cattle market survey of four different regions (Jijel, Constantine, Guelma, and El Tarf). One-hundred percent of the herds were found to be infested in El Tarf and 61% of animals were infested. Within a herd, the intensity of infestation decreased with the age of the cattle. The mean number of warbles per animal decreased as reinfestations occurred. The peak of the emergence of warbles observed between January and March was progressively delayed in the older animals. Two species of Hypoderma were present in cattle in Algeria (H. bovis and H. lineatum) with a predominance of H. lineatum (63%) in the semi-arid area (Batna) and a majority of H. bovis (75%) in humid area (Jijel). Warbles appeared earlier in Batna (October) than in Jijel (November). The Prim'Holstein breed was more susceptible to the warble fly infestation than Montbeliard or local breeds. The implementation of a control programme for hypodermosis in Algeria is urgent. Control can be achieved through a mass treatment of cattle. This approach must take into account the factors described in this study which influence the epidemiology of the disease.