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1.
Fasciola hepatica metacercariae were purified in high yield, removing contaminating cyst walls and plant material by step gradients consisting of 10 ml of 60% Percoll (density = 1.08 g ml-1) and 10 ml of 50% Metrizamide (density = 1.25 g ml-1). Greater than 90% of the metacercariae applied to the density gradients were recovered. These isolated metacercariae had an in vitro excystment rate of greater than 80%, which was the same excystment rate as metacercariae not subjected to density gradient centrifugation.  相似文献   

2.
An enzyme-linked immunosorbent assay (ELISA) was evaluated as a method of determining relative immunoconversion rates in calves and how immunoconversion rates and strength of optical density values correlate with prevalence of Fasciola hepatica fecal egg shedding. Ten to 55 calves and cows were examined from each of 10 separate beef cattle herds in central and southern Louisiana. Infection prevalence rates for calves averaged 8% higher when ELISA optical density values were used than those when fecal egg count data were used. Of 55 calves in 8 herds that were ELISA positive, 39 were shedding F hepatica eggs; of 53 calves that were shedding eggs, 14 were ELISA negative. Significant correlation of calf fecal and ELISA prevalence was observed for 8 herds by linear regression analysis. A chi 2 analysis showed that calf ELISA and fecal egg shedding data were not independent. Results indicate that positive ELISA reactions for as few as 10 to 15 calves from the fall calf crop of a given herd are sufficiently accurate to be used to assess F hepatica herd infection rates, the likelihood of liver condemnations at feedlot destinations, and variation between individual farms in fascioliasis infection risk. The test was less valuable as a diagnostic test when used in adult animals previously exposed to F hepatica or on an individual animal basis.  相似文献   

3.
A coprological sedimentation method is evaluated for quantification of egg shedding in bovine faeces. Through the inclusion of different number of Fasciola hepatica eggs in negative faeces, egg recovery rate and the sensitivity of the method were determined. The mean egg recovery rate of the technique was 76.72+/-15.42%. The sensitivity of the method was 33.3% whenever eggs/g of faeces (EPG) are less than 1.5 and 100% for higher values. To improve the diagnostic accuracy with this technique, it is advisable to increase the sample from 10 to 30g of faeces when manipulating low egg shedding, which allowed for a sensitivity of 83.3%. Regression equations were calculated to quantify the relationship between the number of recovered and incorporated eggs.  相似文献   

4.
We have evaluated the efficacy in sheep of a combination drench formulation at the recommended dose rate of 0.2 mg moxidectin/kg bodyweight and 10 mg triclabendazole/kg bodyweight against an experimental infection with Fasciola hepatica and a natural infection with gastrointestinal nematodes. We confirmed that the efficacy of reducing fecal egg output was 98.3% for trichostrongyle eggs and 100% for F. hepatica eggs. Based on adult worm and fluke recovery, the efficacy varied according to the target species. A reduction was found in the number of Teladorsagia circumcincta, Trichostrongylus spp., Nematodirus spp., and Trichuris spp. greater than 95%, but the efficacy for Oesophagostomum spp. varied, with values below 90%. The reduction in F. hepatica was higher than 95% for all stages. The effectiveness of the formulation was also confirmed by an increase in total proteins and albumin following treatment.  相似文献   

5.
A method was developed for percutaneous ultrasound-guided cholecystocentesis in cattle. The procedure was performed on the right side in the 9th, 10th, or 11th intercostal space of 30 cows. Of the 30 cows, 20 were slaughtered 24 hours after cholecystocentesis and the remaining 10 cows were slaughtered after a 10-day observation period. Changes in the peritoneum and gallbladder wall, observed at slaughter, were minimal. During the 10-day observation period, general behavior, attitude, and appetite of the 10 cows were normal. A transient, slight increase in rectal temperature was observed in 6 cows at 4, 5, or 8 days after cholecystocentesis. Total and differential WBC counts and total protein and fibrinogen concentrations, determined daily, were all within normal ranges. Bile samples from 20 cows were examined microscopically and biochemically. Fasciola hepatica and Dicrocoelium dendriticum eggs were observed in bile from 7 and 12 cows, respectively. Fecal examination revealed F hepatica eggs in 4 cows; D dendriticum eggs were not identified in any of the fecal samples. In 1 cow, F hepatica eggs were observed in the feces, but not in the bile. Bile acids concentration in bile varied from 12.5 to 68.5 mmol/L (mean +/- SD, 45.3 +/- 3.05 mmol/L) and in serum from 3.8 to 281.0 mumol/L (41.6 +/- 17.24 mumol/L). Negative correlation was obtained between bile acids concentration in bile and that in serum (r = -0.60, P less than 0.01). It was concluded that percutaneous ultrasound-guided cholecystocentesis in cows is a safe procedure and that microscopic and biochemical examinations of obtained bile can be useful diagnostic aids.  相似文献   

6.
The results of a study testing the susceptibility of the hispid cotton rat, Sigmodon hispidus, to infection with Fasciola hepatica are described. Ten cotton rats were placed into one of four test groups, one uninfected control or three animals each receiving two, five, or 10 F. hepatica metacercariae. Three of nine (33%) became infected, and two of these animals carried the infection to patency. At necropsy one animal had two mature F. hepatica within the common bile duct, and the other harbored a single fluke. Gross and microscopic changes, primarily of the liver, associated with infection are described, as are fecal egg counts throughout the duration of infection.  相似文献   

7.
The highest prevalences and intensities of human fasciolosis by Fasciola hepatica are found in the northern Bolivian Altiplano, where sheep and cattle are the main reservoir host species and pigs and donkeys the secondary ones. Morphometric comparisons of many linear measurements, areas and ratios of F. hepatica adults (from sheep, cattle and pigs) and eggs (from sheep, cattle, pigs and donkeys) in natural liver fluke populations of the Bolivian Altiplano, as well as of F. hepatica adults and eggs experimentally obtained in Wistar rats infected with Altiplanic sheep, cattle and pig isolates, were made using computer image analysis and an allometric model. Although morphometric values of adult flukes from natural populations of sheep, cattle, and pigs showed great overlap, there were clear differences in allometric growth. The allometries analyzed were: body area (BA) versus body length (BL), BA versus body width (BW), BA versus perimeter (Pe), BA versus distance between posterior end of body and ventral sucker (P-VS), BL versus BW, BL versus Pe, and BL versus P-VS. These allometries show a good fit in the seven pairs of variables in all the populations examined. Comparative statistical analysis of the allometries shows that fluke adult populations from sheep, cattle and pigs significantly differ in BL versus BW and BL versus P-VS functions. Statistical analysis of F. hepatica egg size shows characteristic morphometric traits in each definitive host species. In experimentally infected rats, fluke adult allometry and egg morphometry do not vary depending on the Altiplanic definitive host species isolate. Our study reveals that the definitive host species decisively influences the size of F. hepatica adults and eggs, and these influences do not persist in a rodent definitive host model.  相似文献   

8.
We evaluate the ability of a Fasciola hepatica FABP native antigen (Fh12) with a new vaccination system called ADAD to protect mice and sheep against an experimental F. hepatica infection. The vaccination protocol consists of a set of two injections. The first injection contains a micelle in which two components are included, saponin from Quillaja saponaria (Qs) and/or Anapsos (A) a Polypodium leucotomos hydroalcoholic extract, both emulsified in a non-mineral oil (Montanide) in a water/oil emulsion (30/70). This is subcutaneously injected to achieve the "adaptation" of the immune system to subsequent stimuli. The second injection contains in addition the Fh12 antigen. Two different experiments were carried out using two mouse strains (BALB/c and CD-1). Mice vaccinated with Qs+A+Fh12 presented a survival rate of 40%, when compared with control groups. Furthermore, we evaluated the efficiency of the vaccination in sheep against an experimental F. hepatica challenge. The vaccinated sheep presented lower fluke recovery (24.5%), number of eggs in bile fluid (58.1%) and faeces (40.3%) than control groups. The recovered flukes were shorter (32.7%), immature (34.0%) and with lower body mass (31.6%) than non-complete vaccinated sheep. Thus, the new ADAD system could be a good alternative for future vaccination experiments against fasciolosis.  相似文献   

9.
The objective of this study was to evaluate selected morphological characteristics and the chemical composition of turkey eggs of different weight classes. The percentage of eggs weighing ≥100 g increased to 46.66% in wk 23. Eggs weighing 95 and 110 g had lower yolk content (3.05 and 4.86%) and higher albumen content (by 3.16 and 5.21%) compared with eggs weighing 80 g. The total protein content of the albumen increased (from 9.93 to 10.71%) and the water content of the albumen decreased (from 88.30 to 87.59%) with an increase in egg weight. Albumen weight per gram of yolk increased from 1.87 to between 2.18 and 2.42 g in larger eggs. The energy density of the egg decreased from 5.68 to 4.90 kJ/g with an increase in egg weight, and the energy density of the yolk remained at a similar level in all egg weight classes. The other analyzed yolk and shell parameters were not affected by egg weight. The highest energy and total protein content of the yolk was observed in wk 14 of the laying season. No correlations were found between the progress of the laying cycle and eggshell percentage and the water content of the yolk. The values of the remaining egg parameters varied over the laying season.  相似文献   

10.
Diagnostic antigens in bile and feces from Fasciola hepatica infected cattle were detected and characterized by enzyme-linked immunotransfer blot (EITB) techniques. As sources of antigen, samples of bile, intestinal contents and feces were collected from five uninfected calves and from 10 calves with known Fasciola hepatica burdens. A band detected by EITB using a densitometer in the area corresponding to 26 kDa reacted with rabbit anti-fresh fluke antigen and infected cattle sera but not with fluke-negative rabbit sera, rabbit anti-Fasciola hepatica egg sera, Fascioloides magna positive or negative cattle sera. This band was not detected by Coomassie blue in sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) gels or by Ponceau-S stained nitrocellulose strips. Band groups located at 104-66, 66-42, 42-26 and 25-16 kDa reacted inconsistently with the above sera. Sera from mice hyperimmunized with Fasciola hepatica excretory-secretory (ES) products detected only the 26 kDa band by EITB, without cross-reactivity with bands in the other molecular weight (MW) ranges. The results suggest that the 26 kDa antigen may consist of a stable component of ES products and/or tegument-related worm antigen. Diagnosis of Fasciola hepatica through detection of specific, stable antigens in feces of infected animals offers potential advantages over serum-based tests of better sample accessibility, discrimination between previous and current infections, and possible semi-quantitation of fluke burdens.  相似文献   

11.
Sera from 100 herds of cattle located in the state of Washington were examined with an enzyme-linked immunosorbent assay for antibody to Fasciola hepatica in a screening procedure that included 5 to 10 samples/herd. Twenty-eight herds contained infected cattle and F hepatica was most prevalent in 3 distinct geographic areas. Subsequent retesting of all sera available from 14 herds (mean of 109 samples/herd) revealed that the screening procedure correctly detected 7 of 7 operations in which greater than 40% of samples were positive or suspect and 3 of 3 operations in which 12% to 13% of the samples were positive or suspect. One of 3 herds considered negative after screening was found to contain a few (7%) positive samples and 1 herd considered possibly infected was negative on retest. These results were compared with those obtained by fecal examination for F hepatica eggs in 9 of the 14 herds. A good correlation (5 of 5) was found in which a high percentage (48% to 85%) of sera were positive or suspect. Fasciola eggs were not found in samples from 2 herds with few (7% to 12%) positive or suspect sera or in 2 herds that were negative by an enzyme-linked immunosorbent assay.  相似文献   

12.
Circulating antibody against Fasciola hepatica antigens was determined by enzyme-linked immunosorbent assay (ELISA) and immunoelectrophoresis in alpacas naturally exposed to F. hepatica. Serological assay parameters were established by using sera from eight infected animals and seven controls with no record of this parasitic infection. Excretory--secretory (ES-) products, Fas1- and Fas2-ELISA were used to survey 307 alpacas from a F. hepatica endemic area in the Peruvian Andes. Seroprevalence of F. hepatica infection varied from 56.7, 64.8 and 66.8% measured by Fas1-, Fas2- and ES-ELISA, respectively. The sensitivity for ES-ELISA was 95%, corresponding Fas1- and Fas2-ELISA sensitivity values were 90 and 95%. In this population, 7% of animals were positive for F. hepatica eggs in faeces, other parasites detected were Trichuris sp. (40%), Nematodirus sp. (34.6%), Lamanema sp. (12.8%) and Eimeria sp. (11.8%). The results show that F. hepatica infected animals elicit circulating antibodies against ES, Fas1 and Fas2. Fas2-ELISA may be proposed as a sensitive assay for the immunodiagnosis of fasciolosis in alpacas.  相似文献   

13.
A controlled trial was conducted to evaluate the current efficacy of albendazole (ABZ), rafoxanide (RFX), triclabendazole (TRC) or clorsulon (CLS) against Fasciola hepatica in naturally infected cattle. This trial was conducted in Turkey during the spring, the time of year when liver fluke infection is endemic. Fifty crossbred cattle were selected for inclusion in the trial based on finding eggs of F. hepatica in the feces. The cattle were weighed and randomly allotted into five groups of 10 cattle and treatments were as follows: Group 1 served as non-treated control (CONT), Group 2 was treated orally with ABZ at 12 mg/kg, Group 3 was treated orally with RFX at 10 mg/kg, Group 4 received TRC orally at 12 mg/kg and Group 5 received CLS administered subcutaneously (s.c.) at 2 mg/kg. On day 0 (inclusion day), individual fecal samples were collected on days 0 (inclusion day), 7, 14, 28 and 56, after treatment. The drug efficacy was assessed as a percentage of the egg or fluke reduction and body weight gain relative to the untreated control. The results in the study showed a mean reduction of egg counts by 66.7%, 68.2%, 78% and 84.2% in Groups 2-5, respectively. In conclusion, our results indicate that CLS is a highly effective compound for the treatment of F. hepatica in cattle under these field conditions.  相似文献   

14.
1. Successful invasion by nematode parasites is associated with several factors including egg hatching at the right time in their hosts. To determine a simple and appropriate medium for culture and egg hatching of the highly pathogenic species of the Acuariidae family, Cheilospirura hamulosa were cultured in three different media. In addition the viability of C. hamulosa eggs was determined after storage in frozen infected gizzards.

2. Eggs removed from the uteri of the female worms in infected gizzards were pooled and washed in distilled water and screened under a stereo dissecting microscope. Eggs were counted and cultured in three different media, nutrient agar, normal saline 0.9% and Bearman, at room temperature. Additionally, 10 infected gizzards were kept at ?20°C for 2 and 8 months.

3. After 4–5 d there had been no growth in the nutrient agar medium, whereas 11% of the cultured eggs in the Bearman medium contained larvae 2–3 d after culturing. In 0.9% normal saline medium the two polar knobs appeared on the two poles of the eggs at 2 d post cultivation, and 74% of the eggs contained a larva on the third day. Mature larvae gradually exited from the eggs.

4. Eggs collected from female worms in gizzards frozen at ?20°C were cultured in the same three culture media at room temperature. Larvae were visible in the eggs after 2–3 d in the Bearman and 0.9% normal saline media and hatched thereafter.

5. The 0.9% normal saline medium is recommended for egg hatching and cultivation of C. hamulosa due for simplicity, efficacy and cost effectiveness. Moreover, freezing of the infected gizzards at ?20°C is proposed for long-term storage of the eggs.  相似文献   

15.
In the winter of 2008, cattle on a farm in the province of Neuquen, Argentina died from subacute and chronic liver fluke disease despite four previous treatments with Triclabendazole (TCBZ). In the spring of 2009, a preliminary efficacy test revealed good performance using nitroxynil, whereas TCBZ efficacy was only 18% by egg counts of Fasciola eggs in the faeces. Resistance to Fasciola hepatica to TCBZ has never been reported in South America, so in January of 2010 a controlled trial was conducted to confirm and to define the degree of resistance in this herd. In a clinical trial, the fluke egg output was monitored on Days 14 and 21 and serum enzymes gamma-glutamyl transpeptidase (GGT) and glutamic-oxaloacetic transaminase (GOT) on Days 0 and 21 in 36 calves treated with TCBZ or with closantel. The results showed a reduction of 100% in fluke egg output at Days 14 and 21 for closantel. The mean epg in the TCBZ-treated groups did not decrease. Because of the fact that in this study TCBZ treatment in cattle had no effect, even at double the recommended dose, it is highly indicative that resistance of F. hepatica against TCBZ is present on this farm. The GGT and GOT levels decreased in the closantel-treated group as a result of the treatment at 21 days after dosing. To evaluate the importance of TCBZ resistance in F. hepatica in Argentina, a study on more farms from endemic areas is needed.  相似文献   

16.
The pattern of in vivo uptake of albendazole (ABZ) and its major metabolite, ABZ-sulphoxide (ABZSO), by Haemonchus contortus and Fasciola hepatica recovered from ABZ-treated sheep, was investigated. Concentration profiles of both compounds were simultaneously measured in target tissues/fluids from the same infected sheep. In addition, the proportion of the (+) and (-) ABZSO enantiomers was determined in plasma, bile and F. hepatica recovered from treated sheep. Sheep naturally infected with H. contortus were intraruminally (i.r.) treated with ABZ (micronized suspension, 7. 5mg/kg) and the plasma concentrations of ABZSO and ABZ-sulphone (ABZSO(2)) determined in addition to the concentration of ABZ and ABZSO in H. contortus, abomasal mucosa and fluid content samples. In addition, F. hepatica artificially infected sheep were treated i.r. with the same ABZ suspension (7.5mg/kg), and samples of blood, bile, liver tissue and adult flukes were collected and analysed by HPLC to determine the concentrations of ABZ and both enantiomers of ABZSO. ABZSO and ABZSO(2) were the analytes recovered in plasma with ABZ and ABZSO present in H. contortus. ABZ was the analyte recovered at the highest concentration in H. contortus and abomasal mucosa, whereas higher concentrations of ABZSO were measured in abomasal fluid content. Only low concentrations of ABZ were detected in F. hepatica and bile, but markedly higher concentrations of ABZ were measured in liver tissue. ABZSO was the main molecule recovered in F. hepatica, plasma and bile samples collected from ABZ-treated sheep. The (+) enantiomer of ABZSO was recovered at a higher proportion in plasma (75%), bile (78%) and F. hepatica (74%) after ABZ administration to infected sheep.  相似文献   

17.
Resistance to thiabendazole (TBZ), fenbendazole (FBZ) and levamisole (LVM) in naturally acquired gastrointestinal nematode parasites in sheep was investigated on a farm where anthelmintic resistance was suspected. This was measured by both the in vitro egg hatch assay, and reductions in faecal egg and worm counts in treated animals. In the egg hatch assay, nematode eggs were incubated in various concentrations of either TBZ or LVM. The level of resistance was expressed as the drug concentration inhibiting 50% of the eggs from hatching (LC50). The nematode population had LC50 values of 0.26 microgram ml-1 TBZ and 3.12 micrograms ml-1 LVM. In the faecal egg and worm count reduction test, naturally infected sheep were treated with either TBZ (88 mg kg-1), FBZ (10 mg kg-1) or LVM (15 mg kg-1). Faecal egg and total worm counts from these sheep were then compared with counts from untreated sheep. TBZ, FBZ and LVM failed to reduce the faecal egg counts and total worm counts by more than 90%. Based on the identification of larvae from faecal cultures, the most predominant nematode species in the resistant population were Haemonchus (62%) and Trichostrongylus (28%). TBZ reduced faecal egg counts for both species by less than 90%. FBZ and LVM also reduced Haemonchus spp. eggs by less than 90%. Other nematode species numbers did not satisfy criteria for the determination of efficacy.  相似文献   

18.
The emission of Fasciola hepatica eggs in faeces is usually subject to oscillations along time in animals as well as humans. Thus, looking for alternative biological markers reflecting eggs shed per gram of faeces (epg) with lower oscillations may be useful. This study analyzes the possible relationship between liver-fluke uterus area and epg. Uterus area (UA) development of adult F. hepatica obtained at different days post infection (dpi) in a Wistar rat model with isolates obtained from cattle, sheep, pigs and humans from the endemic human fascioliasis zone of the Northern Bolivian Altiplano was analyzed and compared with the number of eggs shed per gram of faeces as obtained through the Kato-Katz technique. The morphometric study of the UA of liver flukes was carried out using image analysis software. The multiple regression model shows that UA is dependent on dpi and isolate. The evolution of UA vs dpi followed a damped model. This work shows a positive relationship between liver-fluke UA and egg production. The complete absence of eggs in the uteri of some parasite individuals at 300 dpi was observed, which corresponds to the cessation of egg shedding in the advanced chronic stage. The results obtained suggest the necessity to characterize the isolates employed with regard to geographical as well as host origin in fascioliasis studies in which egg production is used as a biological tag.  相似文献   

19.
Objective To describe two cases of infection with Fasciola hepatica i n young farmed emus, subacute and chronic fasci-olosis and a response to treatment of the flock with albenda-zole.
Procedure Gross lesions were found at necropsy and hepatic lesions in microscopic examination. The parasite recovered from one emu was identified by its morphological characteristics and an egg count reduction test was carried out after treatment of the flock with albendazole.
Results Hepatic lesions resembling subacute and chronic fasciolosis of ruminants were identified. An adult fluke was recovered from the liver of one of the birds and was identified as F hepatica . The eggs of the fluke were irregular in shape and size. No fluke eggs were identifiable in faeces of live emus 10 days after treatment of the flock with albendazole at a dose of 10 mg/kg.
Conclusions This is the first reported case of infection with F hepatica i n farmed emus and the first report of the occurrence of Fasciola infection is the class Aves. The irregular shape and size of the eggs may be attributable to infection of an aberrant host. Treatment with albendazole eliminated eggs from the faeces of the flock.  相似文献   

20.
In order to contribute to the knowledge of the epizootiological conditions in which fasciolosis develops in the León mountains, the elimination of Fasciola hepatica eggs in 10% of the total number of cattle (1301 samples) at five locations in the Porma river basin was recorded at monthly intervals between March 1986 and March 1987. The parasite was found in cattle [29.5% prevalence, average of 51.6 +/- 4.5 eggs g-1 (e.p.g.) faeces] throughout the year. The main egg elimination period was winter-spring, with maximum prevalence in January and maximum e.p.g. in February-March. Prevalence of infection in cattle generally increased with the host's age, whilst the average e.p.g. varied between different age groups.  相似文献   

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