ABSTRACT
The study was conducted in Boalia thana under Rajshahi district to identify and analyze the risk factors associated with fascioliosis, during the period of 17th December,2008 to 16th February,2009. In total 34 animals were randomly selected for the study. Among them 22 animals were infestated with fascioliosis. The study showed that fascioliosis was associated with several demographic (sex and age), environmental (grazing area) and managemental factors (farming system and nutritional status). The study showed that animals affected fascioliosis at 1-2 years old (31.82%), 2-3 years old (40.91%) and 3-4 years old (27.27%). The study also revealed that male were at high risk in fascioliosis than female. It was revealed that mixed farming had more importance in infestation of fascioliosis than single farming.
The study showed that malnourished animal were more affected with fascioliosis than healthy animal. It was also revealed that animals reared in highland (27.27%) and animals reared in lowland (72.73%) were infestated with fascioliosis. These result clearly indicated that the managemental factors had more importance in infestation of fascioliosis than other factors.
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Key words: Analysis, Risk factors, Fascioliosis.
CHAPTER-I
INTRODUCTION
Bangladesh is an agrobased country and as a part of agriculture sector livestock have an important role in its economy. Their contribution to GDP is approximately 5.85%.
Bangladesh earns about 13% of total foreign currency by contributing the livestock sub sector (Alam, 1993). There are about 2 crore and 29 lac caltle, 12 lac, 60 thousands buffalo, 2 crore, 15 lac and 60 thousands goat and 27 lac, 80 thousands sheep in this country. But most of the animals are unhealthy, emaciated and their productive performances are not satisfactory due to malnutrition and diseases.
Parasitism is one of the most vulnerable causes of livestock diseases which claims to be the main obstacle in livestock rearing in Bangladesh. (Bhuiyan,1970). The endoparasitic infestation was common problem in this country & causes substantial economic loss. (Rahman and Razzak, 1973; Qadir, 1974; Garrels, 1975).
In Bangladesh among the parasitic diseases Fascioliosis is most important. It is caused by Fasciola gigantica (Bhuiyan, 1970; Amin & samad, 1988; Islam & samad, 1989).
A vast number of livestock population were infected with fascioliosis. Bhuiyan (1970) reported that about 21-22.4% Fasciolioiss in cattle and 12.92 % Fascioliosis in goat were found in Bangladesh.
In Asia and Africa tropical fascioliosis is regarded as of the most important single helminth infections in ruminants (Boray, 1985; Fabiyi, 1987; Murrell 1994; Anon 1995; Harrison et al., 1996; Roberts and Suhardono, 1996; Malone, 1997). Together with major nematode infections, fascioliosis is a signiticant constraint on the productivity of domestic ruminants throughout Asia, South- East Asia and Africa and is thus a significant impediment to global food production (Dargie, 1987; Fabiyi, 1987; Murrell, 1994). Fascioliosis is a major constraint to agricultural and animal production in most countries located in the wet tropics (Spithill et al., 1999). It causes a significant reduction in meat and milk production, growth rate, fertility and draught power in infected animals (Dargie, 1987). From 1975 to 1997, it has been estimated that economic losses in tropical countries reached more than US $ 3200 million
(Spithill et al., 1999). In Bangladesh, Bhuiyan (1970) reported that the economic loss caused by Fasciola gigantica infection in domesticated ruminant in terms of liver damage was taka 3.2 million. The total loss due to Fascioliosis world wide was accounts to two thousands million US dollar per year (Boray, 1985).
Fasciola gigantica requires a suitable place and environment for spreading it’s infection, that’s why the epidemiology of Fasciola gigantica is very important.
Arambulo and Moran (1981) reported that the climatic condition and geographic position of locality influence the type and severity of parasitic infestation in grazing animals. Fasciola gigantica found in low land tropical areas. and several factors are responsible for its infection these are known as risk factors. These are availability of intermediate host snail (Lymnea auricularia), temperature, moisture, season, sex, parity, health status of animal, availability of flooded area or irrigated land which are used as grazing land of animal. A very few limited studies were done on this topic.
For this reason the present study was conducted with the following objective:
i) To identify and analyze the risk factors for fascioliosis.
CHAPTER-II
REVIEW OF LITERATURE
The life cycle and risk factors of Fascioliosis are presented below:
Fig-1: Life cycle of Fasciola gigantica
Risk factors:
1. Availability of host of parasite:
All ruminants are affected by Fasciola gigantica and Fasciola hepatica (Chakrabarti, 1998). In Bangladesh there was an investigation on fascioliosis in which 60% cattles, 90.9% Buffaloes, 19.92% goats and 8.4% sheep were infested with
Fasciola gigantica (Bhuiyan,1970).
2. Availability of intermediate host:
Where a parasite requires intermediate host, its distribution is greatly influenced by availability of intermediate host (Rahaman and Hafezur, 1996).
In South, West and East Africa Lymnaea natalensis is the intermediate host of Fasciola gigantica whereas Lymnaea auricularia rufescens is intermediate host of Fasciola gigantica in Bangladesh, India and Pakistan (Kendall, 1954). Lymnaea ollula is host to Fasciola gigantica in Japan (Ueno et al., 1975). The intermediate host of Fasciola gigantica live deeper water and are close to being true aquatic snails in there behaviors ( Hansen et al., 1994). In Nepal, Morel and Mahato (1987) concluded that both Lymnaea auricularia rufescens and Lymnaea luteola act as intermediate hosts for Fasciola gigantica .
3. Climatic condition and Geographical distribution:
The climatic condition and geographical position of locality influenced the types and severity of parasitic infestation in grazing animals (Arambulo and Moran, 1981).
a) Effect of temperature:
On fluke eggs:
The time required for development of miracidia in eggs of Fasciola gigantica varies with temperature. It was 10-11 days at 37-38ºC, 21-24 day at 25ºC and 33 days at 17- 22ºC (Guralp et al., 1964). Grigoryan (1958) considered 24-26ºC pH 6.5-7 optimal.
and found that under such conditions, 70-80% of eggs would develop. Eggs of Fasciola gigantica do not all develop at the same rate sothat from the same batch, miracidia may hatch over a period up to 14 weeks, thus enhancing their opportunity to infect a snail (Guralp et al., 1964).
On snail development:
A mean day/ night temperature of 10ºC or above is necessary for snails to breed and when temperatures rise to 15ºC or above that level, a significant snail multiplication takes place( Urquhart et al., 1996).
On parasite development in the snail:
Grigoryan (1958) considered 24°C to 26°C optimal for development of miracidia, found that eggs did not survive temperatures more than 43°C and desiccation was rapidly fatal. At 27°C cercariae may develop as early as 26 days after snails are infected (Al- Kubaisee and Altaif, 1989). Development of larvae in the snail becomes slower as the temperature drops and it eventually stops. Below 16°C only a succession of daughter redial generations are produced, but they switch to production of cercariae when the mean temperature is raised to 20°C (Dinnik and Dinnik,1964).
Cercariae are shed in up to 15 waves (usually three or fewer) one to eight days apart over a period of up to 50 days (Grigoryan 1958; Da Costa et al., 1994; Dreyfuss and Rondelaud, 1994). Maximum shedding at the optimum temperature range of 25°C to 27°C occurs 46–50 days after infection (Dinnik and Dinnik, 1963; Asanji, 1988;
Sharma et al., 1989). The number of cercariae produced per snail is usually a few hundred, but varies from fewer than one hundred to over a thousand.
On Metacercarial survival:
The duration of survival of metacercariae is inversely related to temperature of storage. In Japan, Kimura and Shimizu (1978) showed some metacercariae remained viable over on rice stems in a barn (2-28ºC and relative humidity 37-88%) for 120 days but not 150 days.A few metacercariae survived in lucerne hay for 15 days at ambient temperature of 21-32ºC and relative humidity of 30-50% but all were dead by day 35 (Grigoryan,1959).
A similar result was obtained for survival of metacercariae in silage (Grigoryan, 1959;
Gupta and Kamra, 1987).
Desiccation of metacercariae in a dish was ever more rapidly fatal, all being found dead by day 15 at 24-29ºC (Grigoryan, 1959).
b) Moisture:
Fasciola spp. require moisture for transmission, proliferation and survival; fluke eggs need moisture to break up the fecal mass for their develop; miracidia need wet surfaces to find snail hosts; snails need moisture to develop; cercariae donot normally emerge from snails unless there has been recent rainfall; and metacercariae need humidity to survive (Dalton,1999). The ideal moisture conditions for snail breeding and the development of Fasciola sp. with in snail are provided when rainfall exceeds transpiration and field saturation is attained (Urquhart et al., 1996). In Bangladesh optimum development (egg to miracidium) start at wet season (late June to early July). Development in snail is complete by the end of rainy season, shedding of cercariae from snail start in dry season. and clinical sign occur at the end of dry season (October- November). The habits of the host may allow it to come – in- contact with the infected material thus completing the methods for the completion of the life cycle of a parasite for its survival (Bosak, 1999).
c. Effect of Season:
It is another important factor for infestation. Schillhorn van veen (1980) reported that the population of snails and their prevalence of infection with Fasciola gigantica were highest at the beginning of the dry season, leading to outbreaks of fasciolosis in sheep at the end of the dry season.
Ogunrinade (1985) in Nigeria in an abattoir study which revealed that prevalence of infection in cattle was highest in April to June:
In Pakistan, Swarup and Pachauri (1987) found the highest prevalence of infection of snails after the end of the wet season in October, with larval development in snails during the following spring months followed by infection of stock in summer from March onwards.
Sharma and Lal (1983) reported 15% infestation rate with Fasciola gigantica in buffaloes in Mecrute district of India. The highest incidence was recorded in rainy season.In Bangladesh by Chowdhury et al. (1994) who found the highest prevalence of infeection in snails occured after the end of the monsoon season, from July to Spetember. In Bangladesh Animal infected during dry season and clinical sign occure
at the end of dry season (October- November) (Bosak, 1999). In India Nation wide survey in dairy animals organized by the National Dairy Development Board (NDDB) indicated two critical periods in the year: July- September and February – March (Sanyal and Singh, 1995).
d) Effect of low land, flooded area and irrigated land:
In lowland tropical areas Lymnea auricularia (the intermediate host snail) breeds throughout the year in favourable habitats (Chartier et al., 1990). The population of these snails in rivers may be negatively related to rainfall, reflecting the disruption of the habitat of the snails by flooding and also their dispersion by floodwater, as they spend up to 70% of their time floating at the surface (Widjajanti, 1989). Once water levels stabilise, the population level of snails increases and is most numerous while these conditions persist. The duration and timing of this favourable period for snails may be only a few months at the end of the wet season in closed water bodies, or persist throughout the dry season in slow moving rivers (Mzembe and Chaudhry, 1979; Chartier et al., 1990; Tembely et al., 1995). Once the water level drops too fast during the dry season for the fringing aquatic vegetation to persist or the oxygen level drops too low, the habitat will be rendered unsuitable for these snails (Kendall, 1954).
In irrigated rice fields the population of snails is influenced by the availability of water for irrigation and the stage of growth of the crop. Snails and their eggs surviving from the previous crop may colonize recently planted rice fields or they may enter with water introduced to flood the field after planting. Their numbers then increase over the next few months before declining again a few weeks before harvest (Widjajanti, 1989). Despite being aquatic, the snail hosts of F. gigantica are able to survive desiccation in the shade on the surface of the soil for some weeks. After one month, Widjajanti (1989) observed that mean survival time was reduced by about one-third, and egg masses by about one-half relative to hydrated controls. However, she found no adverse affect of desiccation for one month on subsequent hatchability of eggs. The eggs did not hatch until rehydrated, prompting her to propose this as a possible mechanism for survival of the population in habitats subject to periodic desiccation.
4. Factors related to host:
a) Sex of host:
It may be another factor of fascioliosis. Okiluddin (1996) examined fecal sample of cattle in Kishoregonj district. By fecal examination, the prevalence of fascioliosis in female animal was 26.22% and in male animal 17.55%. The prevalence of fascioliosis was 72% in female and 62% in males in Philippines (Copeman et al., 1999).
b) Age of host:
Young animal are more susceptible to parasitic infection than adult, in goat it range from birth to 1.5 years of old (Blood et al., 1983).
The older host of parasite are resistant than the young host, due to appearance of same physiological activities in the older host that are normally absent in young (Rahman and Hefizur, 1996).
Cattle older than four years had the highest prevalence (82%), for those two to four years old the prevalence was 78% and for those five months to two years, prevalence was 53% (Copeman et al.,1999).
c) Nutritional status of host:
A well nourished animal is more resistance to infection than a poorly nourished animal. Pathogenic effect of parasitic infections in grazing ruminants is manifested due to reduction of voluntary feed intake specially concentrate feeding. (Tibor Kassai, 1999).
Other factor such as stagnant pond bathing or river/ canal bathing, mixed farming of small and large ruminants and pasture grazing also influenced fascioliosis (Kasib et al., 2009).
CHAPTER-III
MATERIALS AND METHODS
Study area and periods of study
This study was conducted in Thana Veterinary Hospital, Boalia Rajshahi. The study was conducted for the period from 17th December, 2008 to 16th February, 2009.
Determination of minimum sample size required for the study
The sample size of 34 animals was determined by the following formula (Cannon and Roe, 1982):
Required minimum sample size, n = [1-(1- a) 1/D] [(N-(D-1)/2] where, n= Required sample size, N= Population size, D = Number of diseased animals, a = Desired confidence interval. By this formula, a population that contains 10000 animals with 10% expected prevalence of fascioliosis; the range of calculated minimum required sample size in 95% confidence interval was 29-44. In this study 34 no. sample were selected by random sampling method.
Case definition, categorization of risk factors and data collection
The feces examination and clinical signs were considered to determine the cases of fascioliosis.
Feces examination
The direct microscopic examination method was used for fecal examination. The direct microscopic examination method is a qualitative test for the detection of eggs in the feces.
Equipment
Slides and coverslip
Microscope
Saline solution
Sample (faeces)
Cotton plug Procedure
1. Taking trace amount of feces in the slide
2. Then mixed trace amount saline water in the slide.
3. Then well mixed the fecal sample with the saline solution in the slide and preparing a smear.
4. The preparing slide fixed under the Microscope.
5. Then observed the eggs in the focus.
Microscopic findings
The chronic fascioliasis can be confirmed by the detection of large number of characteristics operculated fluke eggs in the faeces under the microscope. Fasciola eggs are thin walled and stained yellow-brown by biliary pigments. Operculated fluke eggs are also characteristics of paramphistomum but somewhat larger and don’t stained yellow, have a transparent shell, a much more distinct operculum and well defined embryonic shell.
Clinical signs
Following clinical signs were considered important in diagnosing fascioliosis, such as- inappetance, depression and weakness, loss of body weight, loss of production, diarrhoea, anaemia, submandibular edema (bottle jaw).
Fig-2: Bottle jaw in goat Fig-3: Microscopic examination of feces
Epidemiological information was collected about the possible risk factors for fascioliosis through a preset questionnaire. The risk factors are categorized as: (1) Demographic variables, such as – age and sex. (2) Managemental factors, such as- mixed farming and malnutrition. (3) Environmental factors, such as- grazing on low lying areas and grazing on highland pastures.
Analysis of data
Data were sorted in the Microsoft Excel spreadsheet and analyzed by the computer software STATA 7 according to different variables. The measures of association and strength of association between fascioliosis and possible risk factors were calculated and their significance levels were tested.
CHAPTER-IV
RESULTS AND DISCUSSION
The distribution of different variables among the 34 animals with their frequencies and percentage are presented in Table 1. Among the observed 34 no. of animals, 22 animals were found infested with fascioliosis. The highest rate of infestation was found (41.18%) in the 2-3 years age group (Table 1). The measures of association between fascioliosis and different variables are presented in Table 2. Among the affected animals, 2-3 years age group was highly susceptible to infection (40.91%) than 1-2 years age group (31.82%). The infestation rate was comparatively lower (27.27%) at 3-4 years (Table 2), perhaps due to build up of active immunity. This measures of association was not statistically significant (P=0.98). The chance of infection rate in 2-3 years age group and 3-4 years age group were 1.03 and 1.14 times higher respectively than 1-2 years age group (Table 3). But this was not statistically significant (P=0.89).The analyzed value mismatch with previous information of 78% prevalence in 2-4 years of age reported by Copeman et al. (1999).
Table 1. Distribution of different demographic, managemental and environmental factors
Variables Category Infestation
(%) Non-infestation
(%) Total
(%) Demographic
Age (n=34) 1-2 years 07 (63.63) 04 (36.36) 11 (32.35)
2-3 years 09 (64.29) 05 (35.71) 14 (41.18)
3-4 years 06 (66.67) 03 (33.33) 09 (26.47)
Sex (n=34) Male 18 (78.26) 05 (21.74) 23 (67.65)
Female 04 (36.36) 07 (63.64) 11 (32.35)
Management Farming system (n=34)
Single 10 (50.00) 10 (50.00) 20 (58.82)
Mixed 12 (85.71) 02 (14.29) 14 (41.18)
Nutritional status (n=34)
Malnourished 17 (77.27) 05 (22.73) 22 (64.71)
Healthy 05 (41.67) 07 (58.33) 12 (35.29)
Environmental Grazing area (n=34)
Highland 06 (42.86) 08 (57.14) 14 (41.18)
Lowland 16 (80.00) 04 (20.00) 20 (58.82)
Graph-1: Infestation rate of fascioliosis according to age
Graph-2: Infestation of fascioliosis according to sex
Graph-3: Infestation of fascioliosis according to farming system
63.63, 33%
64.29, 33%
66.67, 34% 1-2 years
2-3 years 3-4 years
78.26
36.36
0 20 40 60 80
Male Female
Male Female
50
85.71
0 20 40 60 80 100
Single farming
Mixed farming
Single farming Mixed farming
Graph-4: Infestation of fascioliosis according to grazing area
Graph-5: Infestation of fascioliosis according to nutritional status
In the sex variable, out of 34 animals, 78.26% male and 36.36% females were infected (Table 1). But among the infected animals, rate of infection exceeded in males (81.82%) than in females (18.18%) and this was statistically significant (P=0.01) (Table 2). The chance of infection in male was 6.30 times higher than female (Table 3) and it was also highly significant (P=0.01). This observation does not support the earlier observation of Okiluddin (1996) and Copeman et al. (1999) who reported higher prevalence of fascioliosis in female.
Table 2. Measures of association between fascioliosis and different demographic, management and environmental exposure variables
Variables Category % of
Infestation
F/chi2 value P value (<0.05) Demographic
Age (n=22) 1-2 years 31.82 (7)
F = 1.000 0.98
2-3 years 40.91 (9) 3-4 years 27.27 (6)
Sex (n=22) Male 81.82 (18) F = 1.000 0.01 *
Female 18.18 (4)
Managemental Farming system (n=22)
Single 45.45 (10) F = 0.066 0.03*
Mixed 54.55 (12)
Nutritional status (n=22)
Malnourished 77.27 (17) Chi2 = 4.310 0.03*
healthy 22.73 (05) Environmental
Grazing area (n=22)
High land 27.27 (6) F = 0.036 0.02*
Low land 72.73 (16)
* (Statistically significant)
In the farming system variable, out of 34 calves, 20 calves were reared on single farming system and 14 calves were reared on mixed farming system and the higher percentage of infestation (85.71%) occurred in animals, which reared on mixed farming system than those reared on single farming system (50.00%) (Table 1).
Among the infected animals, the higher infection (54.55%) occurred in animals reared on mixed farming system than those reared on single farming system (45.45%) (Table 2). This was statistically significant (P=0.03). Mixed farming system can cause higher rate of infestation of fascioliosis which supports the earlier observation of Kasib et al.
(2009).
Table 3. Strength of association between fascioliosis and related risk factors (At 95% confidence interval)
Variables Category Odds Ratio Chi2 value P value (<0.05)
Age 1-2 years 1
2-3 years 1.03 0.02 0.89
3-4 years 1.14 0.00 0.97
Sex Male 1 5.55 0.01*
Female 6.30
Farming system Single 1 4.46 0.03*
Mixed 6.00
Nutritional status
Healthy 1 4.18 0.04*
Malnourished 4.76
Grazing High land 1 4.83 0.02*
Low land 5.33
* (Statistically significant)
The rate of infestation was higher (77.27%) in malnourished animals than those in healthy animals (22.73%) (Table 2). This measures of association was statistically significant (P=0.03). The chance of infestation was 4.76 times higher in the group of malnourished animals than the healthy group of animals (Table 3). It was also found statistically significant (P=0.04). This result agreed with Tibor Kassai (1999).
Grazing on low lying areas is an important predisposing cause of fasciola infestation.
In this study 58.82% calves were found grazing on low land areas and among them 80% animals were infested by Fasciola but 57.14% animals were not infested (Table 1) which grazed on high land areas. Among the affected animals, 72.73% animals were found affected which grazed on low land areas. This infestation is significantly higher (P=0.02) than the animals (27.27%) grazed on high land areas (Table 2). It was probably because the abundant exposure of metacercariae during grazing on low lying areas. The chance of infestion was 5.33 times higher in low land grazing areas than those in high land grazing areas (Table 3). This measures of association was also statistically significant (P=0.02).This observation agreed with the observation of Mzembe and Chaudhry (1979), Chartier et al. (1990) and Tembely et al. (1995).
CHAPTER-V CONCLUSION
From the present study it can be concluded that among the risk factors management factors such as- mixed farming system and malnutrition were the main causes of fasciola infestation. Environmental factor, that is, grazing on low lying areas predisposes to cause infection Overall, all these risk factors play significant role in causing fascioliosis in ruminants.
LIMITATION
1. The study period was short (two months) and it is the main limitation of study.
2. The sample size was small. It was another limitation of the study
ACKNOWLEDGEMENTS
All praises are due to almighty Allah, who enabled the author to complete the study successfully.
The author would like to express my deepest sense of gratitude and thanks to his internship supervisor Dr. Pankaj Chakraborty, Lecturer, Department of Medicine and surgery, Chittagong Veterinary and Animal Sciences University for his scholastic guidance, constant inspiration, sympathetic supervision & valuable suggestion during the study period & also preparing this clinical report.
The author would like to express his cordial thanks to his internship coordinator Md.
Asraf Ali Biswas, Associate Professor, Dept. of Animal sciences and Animal Nutrition, Chittagong Veterinary and Animal Sciences University, for his most valuable guideline about report writing and overall co-operation during the period of this study.
The author is also grateful to Vice Chancellor Dr. Nitish Chandra Debnath specially for giving her chance of working in the field level.
Special thanks to Dr. Kobir Uddin Ahmed Veterinary Surgeon at Boalia, Rajshahi for his sympathy & outmost help, guideline during study.
The author finally, is ever indebted to her family members, friends and all well wishers.
The author
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APPENDIX
Questionnaire for risk factor analysis of fascioliosis
Sl. No. Date:
1. Name of the name:
Address:
2. Description of the animal:
a.Species: b. Breed: c. Age: d. Sex: e. BCS:
3. Clinical sign:
a. Temperature: b. Pulse: c. Respiration:
d. Appetite: f. Body coat: g. Rumination:
h. Feces: i. Urine:
4. First signs appeared:
5. Duration of illness:
6. Clinical signs:
7. Deworming :
8. Vaccinated: Y/N If Yes ……….
9. Feeding: Item a. Roughage b. Concentrate c. Other 10. Housing type: a. Intensive b. Semi Intensive c. Other 11. Type of floor of the House:
12. Environment: (Hygienic / Unhygienic) 13. Breeding history: a. Natural b. AI
14. System affected: a. Digestive b. Respiratory c. Cardiovascular d. Urinary e. Genital f. Integumentary 15. Tentative Diagnosis:
16. Previous treatment:
17. Treatment:
Signature of owner: Signature of Interviewer:
RISK FACTOR ANALYSIS OF FASCIOLIOSIS
A Clinical report submitted by
Examination Roll No: 2003/42 Reg. no: 138
Internship ID: H-40 Session: 2002-2003
This clinical report submitted for partial fulfillment of the degree of Doctor of Veterinary Medicine (DVM)
Faculty of Veterinary Medicine
Chittagong Veterinary and Animal Sciences University.
Khulshi, Chittagong-4202
June, 2009
RISK FACTOR ANALYSIS OF FASCIOLIOSIS
