Liver fluke in Irish sheep: prevalence and associations with management practices and co-infection with rumen fluke.

BACKGROUND: The present study aimed to identify the national prevalence of Fasciola hepatica in Irish sheep and to conduct a risk analysis assessment based on management and treatment practices in participating flocks. Also, co-infection with rumen fluke was quantified and its association with liver fluke and management practices was assessed. METHODS: A total of 305 sheep flocks were selected ensuring even national representation of the sheep population. Participating farms were asked to complete a survey questionnaire on farm management practices and submit faecal samples during the winter of 2014-2015. Pooled faecal samples were analysed for the presence of F. hepatica and co-infection with rumen fluke. Apparent and true prevalence were calculated, additionally, the rate of co-infection with rumen fluke was also obtained. Correlation and regression analyses were used for assessing associations between management practices, liver fluke infection and co-infection with rumen fluke. RESULTS: The national true prevalence of F. hepatica was 50.4% (n = 305). Regional prevalence varied from 41% in the east to 52% in the south. Co-infection with rumen fluke was observed in 40% of the studied population and correlated with increased F. hepatica egg counts (OR = 2.9; P ≤ 0.001). Predominant breeds were Suffolk, Texel and Horned Mountain breeds. Beef cattle were the most frequent type of other livestock present on farms and mixed species grazing was frequently reported (73%). More than half of the flocks reported a mid-to-late lambing period (March-April). Use of mountain land for grazing was of 32%. Flukicides were most commonly used twice over the autumn-winter period. Regression analyses highlighted significant association of F. hepatica status, with the presence of other livestock on farm, frequency of flukicides used during the winter and clinical presentation of liver fluke. A significant increase in eggs per gram of faeces was observed in Charollais sheep in comparison with all other breeds. Co-infection with F. hepatica and Calicophoron daubneyi was also significantly associated with the presence of other livestock on the farm, type of flukicide used and clinical fasciolosis. CONCLUSIONS: The present study provides up-to-date information on the prevalence of F. hepatica in Irish sheep and adds insight to the epidemiology of the disease. These findings will be useful for designing new holistic control measures for F. hepatica infection.

Study on the Relationship between Liver Parasitic Infections and Serum Vitamin A and ß-Carotene Status in Cattle.

OBJECTIVE: This study was conducted to evaluate the relationship between serum levels of vitamin A and ß-carotene in the liver of cattle. METHODS: A total number of 150 samples were selected according to the type of parasitic infections of the liver after postmortem examination and confirmation. Parasitic lesions in the liver were subdivided into three major parasites, including Fasciola hepatica, Dicrocoelium dendriticum, and hydatid cysts. Fifty samples were obtained from cattle without any parasitic infection as a control group. Serum levels of vitamin A, ß-carotene concentrations (µg/dL), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) activity were assayed. RESULTS: Naturally infected cattle with D. dendriticum, F. hepatica and hydatid cyst showed lower vitamin A levels. Serum ß-carotene levels were significantly decreased in all groups (p<0.05). ALT and AST activities in animals with parasitic diseases were statistically higher than in control group (p<0.01). A significant negative correlation was determined between the ß-carotene, vitamin A levels and enzyme activities of the liver in all the three types of liver infections (p<0.001). CONCLUSIONS: This study indicated that serum levels of vitamin A and ß-carotene decline was present in cattle with liver parasite infection and vitamin supplements should be supplied.

Pochonia chlamydosporia in the biological control of Fasciola hepatica in cattle in Southeastern Brazil.

Biological control with the use of nematophagous fungi has been described very successfully by many authors and presents itself as a complementary control method, acting on the free-living forms of helminths. The efficacy of a formulation containing the fungus Pochonia chlamydosporia in controlling Fasciola hepatica eggs in faeces was evaluated in an experimental field assay. Two bovine groups (six animals each) were used: A (control) and B (treated with fungus). At 30 days after deworming, the animals were separated into two similar paddocks with flooded areas and were given pellets containing 25 % mycelial mass (group B) or no fungus (group A) at a dose of 1 g/10 kg body weight, twice a week, during 18 months. Faecal samples were harvested fortnightly in the animals of groups A and B and they were submitted at examination of quantitative sedimentation. The mean count of F. hepatica eggs per grams of faeces was significantly higher in group A (1.19) compared with those from group B (0.82) (P < 0.01). After 18 months, animals from group B had gained 42.33 kg above (17.82 % more by weight) (P < 0.01), compared with the control group (A). Every month, faecal samples from paddocks A and B were collected and they were incubated. P. chlamydosporia was identified only in sample source of the paddock B. It can be concluded that the application of this fungical formulation with P. chlamydosporia 25 % mycelial mass was effective in reducing the availability of eggs in the environment and reinfections in calves in natural conditions.

[Increase of analytical sensitivity of FasciDIG system for the diagnosis of Fasciola hepatica]. / Incremento de la sensibilidad analítica del sistema FasciDIG para el diagnóstico de Fasciola hepatica.

INTRODUCTION: fasciolosis is an endemic disease in cattle in Cuba and there is an increase in the number of reported human cases in recent years. The coproparasitological diagnosis of fasciolosis has low sensitivity and is hard-working; for that reason, it is important to use immunoenzymatic methods mainly those that can detect this parasite antigens in the feces. A system for antigen detection called FasciDIG, with a reported sensitivity of 10 ng/mL has been developed in "Pedro Kouri" Institute of Tropical Medicine. OBJECTIVE: to increase the sensitivity of FasciDIG through some modifications to this diagnostic method. METHODS: two foul dilutions (concentrations of antigen 1 000 ng/mL- 1.95ng/mL in H20 Tween-20) were evaluated in a simulated system using FasciDIG and modified FasciDIG. The FasciDiG was modified using the secondary antibody obtained from rabbit against excretory-secretory antigens of Fasciola hepatica combined with biotin and then adding commercial conjugated extravidine peroxidase. Feces were collected from the rectum of 96 animals for slaughter and were evaluated by both methods, FasciDig and modified FasciDig. Kappa index was calculated between both assays. RESULTS: the detection limit for the FasciDIG was 3.9 ng/mL whereas the modified FasciDIG detected up to 1.95 ng/mL. The agreement index calculated between the two tests was 0.6238 corresponding to an index of substantive or good agreement. CONCLUSIONS: the modified method is more sensitive than FasciDIG and it can supplement the diagnosis of fasciolosis. The number of analyzed samples should be increased and the sensitivity and specificity should also be determined using the serial conic-cup sedimentation technique as the gold standard.

Bioensaio de atividade moluscicida adaptado para a avaliação de extratos de plantas medicinais / Bioensayo de actividad moluscicida adaptado para evaluación de extractos de plantas medicinales / Bioassay of molluscicidal activity adapted for the evaluation of medicinal plant extracts

Nesta nota é apresentada detalhadamente a metodologia (preparação dos extratos, adaptação dos caramujos, ensaio de atividade, destino dos caramujos) usada para a avaliação da atividade moluscicida de extratos de plantas frente a caramujos da espécie Biomphalaria glabrata. A adaptação desta metodologia tem o propósito de avaliar extratos naturais para a busca de produtos alternativos mais baratos, biodegradáveis, seguros e disponíveis localmente, para o controle das populações de caramujos.

The methodology (extract preparation, adaptation of the snails, activity test, destiny of the snails) used for the evaluation of the molluscicidal activity of plant extracts in relation to snails from the Biomphalaria glabrata species appears in detail in this note. The adaptation of this methodology has the purpose of evaluating natural extracts in order to find cheaper, biodegradable, safe and easily available alternative products for the control of the populations of snails.

En este apunte se presenta detalladamente la metodología (preparación de los extractos, adaptación de los caracoles, ensayo de actividad, destino de los caracoles) utilizada para la evaluación de la actividad moluscicida de extractos de plantas frente a caracoles de la especie Biomphalaria glabrata. La adaptación de esta metodología tiene el propósito de evaluar extractos naturales para la búsqueda de productos alternativos más baratos, biodegradables, seguros y disponibles localmente, para el control de las poblaciones de caracoles.

Molluscicidal and antiparasitic activity of Solanum nigrum villosum against Galba truncatula infected or uninfected with Fasciola hepatica.

The present study was based on assessments of the molluscicidal and antiparasitic activities of Solanum nigrum villosum. This plant has been collected in Tozeur's traditional oases (south-western Tunisia). Molluscicidal activities of leaves, ripe fruit and unripe fruit extracts of S. n. villosum have been assessed in experimental and semi-field conditions on uninfected Galba truncatula. Antiparasitic activities have also been assessed on larval stages (rediae, intraredial germinal masses and cercariae) of the parasite Fasciola hepatica infecting G. truncatula naturally. The unripe fruit extract (LC50 = 41.2 mg l- 1) was more toxic than the leaf (LC50 = 132.5 mg l- 1) and the ripe fruit (LC50 = 172.8 mg l- 1) extracts on the snail after 48 h of treatment. One mg l- 1 of cupric chloride produced the death of 97% of G. truncatula after 25 days of treatment. Solanum n. villosum extracts used at the lethal laboratory concentration in semi-field conditions were also toxic to G. truncatula. The unripe fruit extract was more toxic than the leaf and the ripe fruit extracts on larval stages of F. hepatica. Maximum deterioration rates were obtained with the unripe fruit extract: 65.6% for rediae, 77.8% for cercariae and 27.6% for intraredial germinal masses after 48 h of treating the snails. The possibility of the unripe fruit extract application for the control of G. truncatula and F. hepatica larval stages should be considered.

Fasciola hepatica leucine aminopeptidase, a promising candidate for vaccination against ruminant fasciolosis.

Leucyl aminopeptidases (LAP) from different parasitic organisms are attracting attention as relevant players in parasite biology, and consequently being considered as candidates for drug and vaccine design. In fact, the highest protection level achieved in ruminant immunization by a native antigen was previously reported by us, using a purified LAP as immunogen in a sheep trial against fasciolosis. Here, we report the cloning of a full-length cDNA from adult F. hepatica encoding a member of the M17 family of LAP (FhLAP) and functional expression and characterization of the corresponding enzyme. FhLAP was closely related to Schistosoma LAPs, but interestingly distant from their mammalian host's homologues, and was expressed in all stages of the parasite life cycle. The recombinant enzyme, functionally expressed in Escherichia coli, showed a marked amidolytic preference against the synthetic aminopeptidase substrate l-leucine-7-amino-4-methylcoumarin (Leu-AMC) and was also active against Cys-AMC and Met-AMC. Both native and recombinant enzyme were stimulated by the addition of divalent cations predominantly Mn(2+), and strongly inhibited by bestatin and cysteine. Physico-chemical properties, localization by immunoelectron microscopy, MALDI-TOF analysis, and cross-reactivity of anti-rFhLAP immune serum demonstrated that the recombinant enzyme was identical to the previously purified gut-associated LAP from adult F. hepatica. Vaccination trials using rFhLAP for rabbit immunization showed a strong IgG response and a highly significant level of protection after experimental infection with F. hepatica metacercariae, confirming that FhLAP is a relevant candidate for vaccine development.

The presence of Fasciola hepatica (liver-fluke) in humans and cattle from a 4,500 year old archaeological site in the Saale-Unstrut valley, Germany.

During an excavation of a site of the corded ware culture in the Saale-Unstrut-Valley (ca. 3000 BC) in Germany, a soil sample from the pelvis of a human skeleton was studied under palaeoparasitological aspects. Eggs of the trematode Fasciola hepatica and of the nematode genus Capillaria were found. This is the first case of a direct association of a F. hepatica-infestation to both a prehistoric human skeleton and domesticated animal remains. Sheep and cattle bones were present at the same site and F. hepatica eggs were found in bovine samples. This strongly points toward an existing infection cycle, involving humans as a final host.

[Liver fluke infestation in New World camelids. Parasitology, pathology, clinical findings and therapy]. / Leberegelbefall bei Neuweltkameliden. Parasitologie, Pathologie, Klinik und Therapie.

In Llamas and Alpacas infestation with Fasciola hepatica or Dicrocoelium dendriticum can cause liver damage, sometimes even with lethal outcome. Once infected South American Camelids (SACs) react more sensitively to these parasites than other domestic ruminants. We report here on the pathology, parasitology, clinics and therapy of this disease. Concerning Dicrocoelium dendriticum we describe own clinical results and therapeutic outcome in addition to the pathological investigation. According to anatomic corrosion casts, the bile ducts of SACs show more similarity with the equine bile system than with the bile system of domestic ruminants.

[Fasciolinip-1: experimental fasciolicidal activity in sheep]. / Fasciolinip-1: eficacia fasciolicida experimental en ovinos.

The aim of the present study was to evaluate the flukicide efficacy of the 2-amino-5(6)-chloro-1-methoxycarbonylbenzimidazole (II), named Fasciolinip-1, against eight and twelve-week old flukes in sheep. In a first experiment 35 young sheep were infected each with 300 metacercariae of Fasciola hepatica. Eight weeks postinfection five groups of seven animals each were treated as follows: Groups 1, 2 and 3 were given 2.5, 5 and 10 mg/kg of Fasciolinip-1. Groups 4 and 5 remained as non-treated and vehicle controls, respectively. In a second study, 32 sheep were infected each with 150 metacercariae. Twelve weeks postinfection, the animals were divided into 4 groups of 8 sheep each. Groups 1, 2 and 3 were treated orally with 5, 10 and 15 mg/kg of Fasciolinip-1, respectively. Group 4 remained as non-treated control. Fifteen days after treatment, all sheep from both experiments were killed to collect the flukes present in liver. Results indicated efficacies of 19.1%, 49.6% and 65.2% for groups 1, 2 and 3 of the first study and 48.3%, 64.8 and 90.6% for sheep of the second experiment, respectively. It was concluded that Fasciolinip-1 removed 8-week old flukes in limited numbers and 12-week old flukes in 90.6% when used at 15 mg/kg body weight.