Methods for in vitro study of the invasive processes of Fasciola gigantica.

In vitro techniques have been developed to study the invasive processes of Fasciola gigantica. The conditions necessary for excystment were identical with those rquired by F hepatica. The ability of larvae to penetrate mouse gut in vitro was influenced by the composition of the fluid within the gut, and by the region of gut wall to which the larvae were exposed. Larvae remained viable on spleen cell monolayers for at least 60 days.     

Evaluation of a commercially available enzyme-linked immunosorbent assay for detecting antibodies to Fasciola hepatica and Fasciola gigantica in cattle, sheep and buffaloes in Australia

A commercially available ELISA for detecting antibodies to liver fluke was evaluated for use in Australia. Milk and serum samples from cattle and sheep in which infection with Fasciola hepatica was confirmed by detection of eggs in faeces were used to estimate sensitivity. Similar samples collected from cattle and sheep outside the F. hepatica-endemic area were used to estimate specificity. The ELISA was also evaluated for detecting antibodies to F. hepatica in milk from sheep and antibodies to Fasciola gigantica in sera from cattle and buffaloes, but with small numbers of samples. In cattle, the sensitivity and specificity of the ELISA were 98.2% and 98.3% using serum and 97.7% and 99.3% using milk. In infected herds, 41.4% and 41.5% of animals were positive in the serum and milk ELISAs, respectively, whereas F. hepatica eggs were found in faecal samples from 26.5% of animals. In sheep, the sensitivity of the ELISA was 96.9% and the specificity was 99.4%. In infected flocks, 60.2% of animals were positive in the serum ELISA and F. hepatica eggs were found in faecal samples 52.2% of animals. There was perfect agreement in the ELISA between paired serum and milk samples collected from ewes. The assay detected antibodies in sera from cattle and buffaloes with natural and experimental F. gigantica infections. In the experimentally infected animals, antibodies were detected 2 weeks post-infection. We conclude that the ELISA will be a valuable tool for diagnosing F. hepatica infections in cattle and sheep. The assay may also be useful for diagnosing F. gigantica infections but further studies are required to establish sensitivity and specificity.     

The prevalence and economic significance of Fasciola gigantica and Stilesia hepatica in slaughtered animals in the semi-arid coastal Kenya

Fasciola gigantica and Stilesia hepatica constrain ruminant productivity. The prevalence and economic losses caused by F. gigantica and S. hepatica in the ruminant production systems of Taveta division of Kenya were estimated in a retrospective appraisal of the slaughter records on the total number of animals slaughtered and livers condemned over the period 1989 to 2004. Only liver condemnations based on infestation by F. gigantica and S. hepatica were considered for purposes of this study. Liver condemnation rates differed significantly between bovines, caprines and ovines (p≤ 0.05) for F. gigantica (26%, 6.6% and 5.2%, respectively) and for S. hepatica (0.4%, 22% and 28%, respectively). The total loss through condemnation of both F. gigantica and S. hepatica infested livers was 4 408 272 KES (Kenyan shillings) (US$ 72 272). The proportion of loss in bovines, caprines and ovines, was 76%, 17% and 7%, respectively. Fasciolosis contributed 3 505 410 KES (79.5%) and S. hepatica infestation 903 210 KES (20.5%) to the total losses due to liver condemnations. The authors feel strongly that there is a need for more work on the two parasites using live animals to determine the prevalence in animals of various ages, species and breeds and the economics of disease control at farm level.     

Incidence of liver fluke infections (Fasciola gigantica andDicrocoelium hospes) in ruminants in northern Nigeria

Summary One thousand and twenty-four cattle, 550 sheep and 1,748 goats slaughtered in a rural slaughter slab during 1973 to 1975 were examined for evidence of liverfluke infections. The prevalence rate ofFasciola gigantica andDicrocoelium hospes infections was respectively 65.4% and 56.0% in cattle, 40.8 and 13.1% in sheep and 17.6 and 5.2% in goats. Other trematodes detected wereSchistosoma bovis and paramphistomes. The seasonal incidence ofF. gigantica as well as ofD. hospes was highest during and directly after the rainy season. The lower prevalence rate ofF. gigantica, especially in the younger animals, during 1973–1974 was thought to be related to the 1973 drought. This was supported by the low prevalence rate in the long-range trade cattle which originated from drier areas. The results are discussed in relation to the climatic conditions during the survey period, as well as to the difference in epidemiology ofF. gigantica andD. hospes infections in northern Nigeria.

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Incidencia De Infecciones Por Distoma Hepatioc (Fasciola Gigantica Y Dicrocoelium Hospes) En Rumiantes En El Norte De Nigeria

Resumen Con el propósito de estudiar la prevalencia e incidencia de infecciones por fasciola, se examination 1.024 bovinos, 550 ovinos y 1.748 caprinos, en un matadero—laboratorio en el norte de Nigeria. El estudio tuvo lugar en el año 1973 y se prolongó en el 74 y 75. La tasa de prevalencia de las infecciones porFasciola hepatica y Dicrocoelium hospes fue respectivamente 65.4% y 56.0% en bovinos, 40.8% y 13.1% en ovinos y 17.6% y 5.3% en caprinos. Otros tremàtodos encontrados fueronSchistosoma bovis y paranfistomus.La incidencia estacional deF. gigantica y deD. hospes fue alta durante y directamente despues de la estación lluviosa. La tasa de prevalencia más baja deF. gigantica especialmente en animales pequeños, durante 1973 y 74, parece estuvo relacionada con la seuía de 1973. La prevalencia baja encontrada en ganado comercial proveniente de regiones más secas, le dá fuerza a esta hipótesis.Los resultados se discuten en relación con las condiciones climáticas encontradas durante la encuesta y a las diferencias epidemiológicas deF. gigantic yD. hospes en el norte de Nigeria.

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Incidence De La Distomatose (Fasciola Gigantica EtDicrocoelium Hospes) Chez Les Ruminants De Nigeria Du Nord

Résumé 1 024 bovins, 550 moutons et 1 748 chèvres abattus dans les abattoirs ruraux de 1973 à 1975 ont été examinés pour l'étude des affections hépatiques à trématodes. La répartition moyenne des infections àFasciola gigantica et àDicrocoelium hospes a respectivement été de 65,4 p. 100 et 56 p. 100 chez les bovins, de 40,8 p. 100 et de 13,1 p. 100 chez les moutons, et de 17,6 p. 100 et 5,2 p. 100 chez les chèvres. Les autres trématodes mis en évidence ont étéSchistosoma bovis ainsi que des paramphistomes. L'incidence saisonnière deFasciola gigantica tout comme celle deDicrocoelium hospes a été plus élevée durant et directement après la saison des pluies. Le niveau plus bas deFasciola gigantica spécialement chez les animaux les plus jeunes durant 1973–1974 paraît être en relation avec la sécheresse de 1973, de qui semble être confirmé par le taux peu élevé observé chez le bétail de commerce originaire des régions les plus sèches.Les résultats observés sont discutés en relation avec les conditions climatiques dominantes durant la période de l'enquête aussi bien qu'en fonction des différences épidémiologiques des infections àFasciola gigantica etDicrocoelium hospes en Nigeria du Nord.

     

A retrospective study of the prevalence and seasonal variation of Fasciola gigantica in cattle slaughtered in the major abattoirs of Zimbabwe between 1990 and 1999

A retrospective study covering a period of 10 years (1990--1999) was conducted using post mortem meat inspection records of the Veterinary Department Information Management Unit at Harare to determine the prevalence and seasonal variation of bovine fasciolosis in Zimbabwe. Records of monthly and annual returns from five major abattoirs were examined in regard to total cattle slaughtered and the corresponding number of livers condemned due to Fasciola gigantica infection. Prevalence of fasciolosis was calculated as the number of cattle found to be infected with F gigantica, expressed as a percentage of the number of cattle slaughtered. Seasonal variations in the prevalence were examined by pooling respective monthly condemnation data over a 10-year (1990--1999) period. A total of 2,474,232 cattle were slaughtered during this period and 917,565 (37.1%) of these cattle were infected with F. gigantica. The pattern of distribution of F. gigantica was significantly higher in cattle originating from catchment areas of high rainfall than in those of relatively low rainfall, and in those slaughtered during the wet season than those slaughtered during the dry season (P< 0.05). Based on the study findings a control programme for the disease in Zimbabwe is suggested.     

27 kDa Fasciola gigantica Glycoprotein for the Diagnosis of Prepatent Fasciolosis in Cattle

An indirect enzyme-linked immunosorbent assay (ELISA) using 27 kDa glycoprotein of Fasciola gigantica has been evaluated for its potential use in the diagnosis of bovine fasciolosis. Following experimental infection of rabbits, F. gigantica infection-induced antibodies were isolated and later used as ligands in affinity chromatography for isolation of infection-induced antibody-specific proteins. Among the five infection-specific proteins isolated, a glycoprotein of 27 kDa was later isolated by second-step purification using concanavalin A matrix. In crossbred cattle receiving different doses of infection (100, 200 and 400 metacercariae), the anti-27 kDa antibodies were detected as early as the 2nd week post infection. No direct correlation between initial dose, antibody response and fluke establishment was recorded. No cross-reaction was noted with the sera of goats experimentally infected with Paramphistomum epiclitum. ELISA with the 27 kDa glycoprotein could be a feasible diagnostic tool for the early detection of bovine fasciolosis.     

Optimization of the procedure for counting the eggs of Fasciola gigantica in bovine faeces

This paper describes a method for counting eggs of F. gigantica in bovine faeces that optimizes the proportion of eggs recovered and the repeatability of estimates. The method uses 3 g of faeces suspended in 0.05% Tween 20. The suspension is passed through three 6 cm diameter sieves in tandem to remove fibrous debris, with respective apertures of 1 mm, 450 μm, and either 266 or 200 μm. The filtrate is allowed to sediment for 3 min in a conical flask; the sediment is recovered, then resuspended in 200 ml of 0.05% Tween 20 and allowed to sediment. After 3 min the sediment is washed in a sieve with an aperture of 53 μm, which retains the eggs. Eggs suspended in 15 ml of 1% methylene blue are counted using a dissecting microscope. Use of Tween 20 instead of water as the suspending agent for faeces gave a significant threefold increased the proportion of eggs recovered and reduced variability between repeated counts. This method is able to detect about one-third of the eggs present. It was concluded that the high proportion of F. gigantica eggs lost may be due to the presence of hydrophobic and covalent bonds on the eggs that bind them to debris, with which they are discarded.     

Comparative study of sheep and buffalo isolates of Fasciola gigantica in the intermediate host Lymnaea auricularia

A comparison was made of the biological behaviour of two isolates of Fasciola gigantica, obtained from sheep and buffalo, in the aquatic snail Lymnaea auricularia. The sheep isolate exhibited a lower rate of infection, slower larval development and the production of fewer cercariae. Larger eggs, miracidia and metacercariae were obtained from the buffalo isolate while the rediae and cercariae were smaller. Metacercariae of sheep origin were dark brown with a mortality after 80 days of storage at 4 degrees C of 30.6 per cent while those of the buffalo isolate were straw coloured and had a mortality of 10.2 per cent.     

Comparative evaluation of the enzyme-linked immunosorbent assay (ELISA) in the diagnosis of natural Fasciola gigantica infection in cattle

The enzyme-linked immunosorbent assay (ELISA) was evaluated for the diagnosis of naturally acquired Fasciola gigantica infection in cattle in comparison with conventional parasitological techniques. Using unfractionated whole worm extract of F. gigantica as the antigen, it was observed that there was a good correlation between ELISA positivity and positive diagnosis of fascioliasis by post mortem liver examination, bile egg sedimentation and faecal egg sedimentation. There was, however, a disparity between ELISA results and faecal egg flotation results.     

Classification of the parenchymal cells in Fasciola gigantica based on ultrastructure and their expression of fatty acid binding proteins (FABPs)

Parenchymal cells in adult Fasciola gigantica can be classified into three types based on their ultrastructural features and different quantities of fatty acid binding protein (FABP) being stored. Parenchymal cell type 1 (Pc1) has pale cytoplasm consisting largely of a loose network of fine fibers, and it contains few mitochondria but numerous glycogen particles. This cell type may be specialized in the storage and metabolism of glycogen and glucose. Parenchymal cell type 2 (Pc2) has similar cytoplasmic features as Pc1 but contains more numerous mitochondria, and high concentration of FABP as reflected by high density of immunostaining and immunogold labeling using specific monoclonal antibody (MoAb) to FABP as probe. Pc2 may, thus, specialize in the storage and metabolism of fatty acids and other lipids. Parenchymal cell type 3 (Pc3) has dense cytoplasm containing large amount of rough endoplasmic reticulum, Golgi complex and mitochondria, which is typical of a secretory cell. Furthermore, Pc3 has very little glycogen particles and is not stained by MoAb against FABP. It could, thus, be concerned with the synthesis of fibers, which form the scaffold of the parenchyma.     

Molecular prevalence of Coxiella burnetii in milk in Iran: a systematic review and meta-analysis

Tropical Animal Health and Production - Q fever is a major zoonotic disease in the world. The aim of this meta-analysis was to estimate the prevalence of Coxiella burnetii in animal milk in Iran....     

Evaluation of dipstick-ELISA using 28 kDa Fasciola gigantica cathepsin l cysteine proteinase (FgCL3) for serodiagnosis of fasciolosis in naturally infected goats

In the present investigation 100 goats presented for slaughter were used to evaluate the 28kDa cathepsin l cysteine proteinase (FgCL3) dipstick-ELISA for the diagnosis of fasciolosis. Presence of Fasciola gigantica worms in liver at the time of slaughter was taken as a gold standard for the evaluation of the assay. Faeces, blood and liver were taken from all slaughtered goats. Biochemical parameters and coprological examinations using sedimentation technique were also taken into consideration. Total serum proteins, albumin, mean values of aspartate aminotransferase and alkaline phosphatase of positive group were not significantly different from those of the negative group. The diagnostic sensitivity of dipstick-ELISA was higher than coprological detection of fluke eggs. However, specificity was equal for both the tests. The accuracy of dipstick-ELISA in detecting circulatory antibodies during the course of disease was higher than coproscopic examination. In conclusion, the FgCL3 antigen dependent dipstick-ELISA was more reliable and has better scope for field application in the endemic areas.     

Fasciola in horses in the Republic of South Africa: a single natural case of Fasciola hepatica and the failure to infest ten horses either with F. hepatica or Fasciola gigantica

The faeces of 11 horses were examined for Fasciola spp. eggs. One of them was positive for Fasciola hepatica, a finding which was confirmed post-mortem. The 10 negative horses were subsequently infested with either F. hepatica or Fasciola gigantica, each animal receiving orally from 500-9,500 metacercariae. No clinical signs were observed and no fluke eggs were detected in the faeces, and neither immature nor adult Fasciola worms were recovered from the horses slaughtered 16-26 weeks post-infestation with F. hepatica and 28-34 weeks post-infestation with F. gigantica, respectively. The results indicate that horses have a high level of resistance to both South African Fasciola spp. Factors which can probably explain the conflicting reports for the prevalences of fascioliasis in horses throughout the world and the varying results obtained in experimental infestations are briefly considered.     

A comparison of serum biochemical changes in two breeds of sheep (Red Masai and Dorper) experimentally infected with Fasciola gigantica.

Twelve Red Masai and 12 Dorper sheep aged between 6 and 9 months, were acquired from a fluke-free area and sheep of each breed divided into two equal groups of six. Each animal in one group of each breed was experimentally infected with 400 viable metacercariae of Fasciola gigantica. The other groups acted as uninfected controls. Blood samples were taken at weekly intervals for the determination of serum bilirubin, albumin, and gamma glutamyl transferase levels. Following the establishment of infection, albumin levels declined in both breeds of infected animals without any significant difference between the two breeds. However, serum bilirubin and gamma glutamyl transferase (GGT) in the infected animals were elevated significantly more in the Dorper than in the Red Masai sheep. Based on these findings, it would appear that Dorper sheep are more susceptible to the infection than Red Masai sheep.