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Molecular Diagnosis And Therapeutic Trials Against Bovine Fasciolosis In And Around Okara

By: Hafiz Farooq Ahmad (2008-VA-93) | Dr. Muhammad Ijaz.
Contributor(s): Dr. Waseem Shahzad | Dr. Jawaria Ali Khan | Dr. Shehla Gul Bokhari.
Material type: materialTypeLabelBookPublisher: 2015Description: 77p.Subject(s): Department of Clinical Medicine and SurgeryDDC classification: 2256-T Dissertation note: Agriculture sector plays a pivotal role in the economy of Pakistan. At present it participates 11.4% to GDP. Agriculture provides 45% employment to labour of our country and contributes in the development of other sectors of the economy. The livestock sector occupies a distinguishing position in the National Agenda of economic development. It provides net source of foreign income. In the history livestock has been dominated by small holders to meet their needs of milk, food safety and cash earnings on daily basis. Besides, livestock is considered a source of employment generation at rural level. It plays an important role in poverty mitigation and keeps in elevating the socio-economic situation of our rural commonalities. Livestock share approximately 55.4 percent to the agricultural value added and 11.9 percent to national GDP during 2013-14. Anonymous (2013-14). Parasitism is one of the major problems lowering livestock productivity round the earth Vercruysse and Claerebout et al. (2001). The significance of helminthes infections is increased manifold in developing countries like Pakistan where 65.2% population is rural (Population Census Organization (2007) depending upon livestock for their incomes and where parasitism acts as a serious trouble for livestock economy Chaudhry et al. (1984). Among helminthes infections, fasciolosis commonly called as liver fluke disease is of vital importance due to its wider spectrum of ultimate hosts Rondelaud et al. (2001) causing acute and chronic infections Sampaio-Silva et al. (1996). The disease is primarily caused by Fasciola hepatica and Fasciola gigantica Soulsby et al. (1987). Fasciolosis is a parasitic liver infection of wild and domestic ruminants caused by genus Fasciola, which has worldwide distribution Soulsby et al. (1986). Fasciolosis causes economic losses as a result of mortalities, abortions, retard growth, reduced milk and meat production, condemnation of infected liver and emaciated carcasses and cost of animal treatment Gracey and Collins et al. (1992). Fasciolosis caused by F. hepatica is a cause of important economic losses in ruminants. The disease is widely spread throughout in Africa, Asia and the Pacific. Although mostly a problem of young stock, the disease is also found in older animals where it adversely affects health, growth rate and development. Apart from its great veterinary importance throughout the world, fasciolosis caused by both F. hepatica and F. gigantica has recently been shown to be a re-emerging and wide spread zoonosis affecting a number of human populations Mas-Coma and Bargues et al. (1997); Esteban et al. (2003). Prevalence of fasciolosis infestation in buffaloes is 49.01%. The infestation of fasciolosis was optimal from January to September, while least during October to December. Fecal samples indicated F. hepatica prevalence 65%. Whereas prevalence of F. gigantica was observed higher in adult cattle as compared to cattle calves by fecal egg counts Khalil et al. (2009). 4% Buffalo calves infested with trematodes. Availability of intermediate host and the grazing habits of the final host determines the epidemiology and seasonal pattern of infection with trematodes Pfukenyi et al. (2005); Bhutto et al. (2002). The life cycle of F. hepatica includes many stages which develop in the environment or the intermediate host. Metacercariae which is the infective stage encysted and ingested by grazing herbivores. Temperature and rainfall are the important factors affecting the life cycle and the prevalence of F. hepatica infection McCann et al. (2010). Many studies revealed that F. hepatica is endemic in buffaloes, cattle, goats, sheep and humans in Pakistan Ijaz et al. (2009); Iqbal et al. (2007); Khan et al. (2010) and Qureshi et al. (2005). Fasciolosis is the disease by which about 250 million sheep and 300 million cattle are potentially affected worldwide Boray et al. (2005), and more significantly infects all ruminants Iqbal et al. (2007). F. hepatica and F. gigantica are the two liver flukes commonly reported to cause fasciolosis in ruminants Walker et al. (2008). Infected cattle can exhibit Introduction 3 poor weight gain and dairy cattle have lower milk yield, and possibly metabolic diseases Mason et al. (2004). Losses due to fasciolosis in the United Kingdom and Ireland alone are more than £18 million a year Mulcahy and Dalton et al. (2001); €52 million a year or €299 per infected animal in Sweden Schweitzer et al. (2005); 0.26 million USD annual due to fasciolosis associated liver condemnations in cattle slaughtered in Kenya Kithuka et al. (2002). In Pakistan, 35697 USD (3141360 PKR) are reported in only one tehsil of Punjab province, the Sargodha T.U. Rehman et al. (2013). While Fasciolosis is prevalent in whole Punjab province of Pakistan but the marshy areas such as regions of Gujranwala, Lahore, Multan, Faisalabad, Jhang, Muzaffargarh, Sheikhupura, and Sargodha are of major concern. Fascioliasis in ruminants causes substantial economic losses, estimated at US$ 2 billion per annum worldwide, to rural agricultural communities and commercial animal producers due to death of infected animals, condemnation of affected livers and production losses associated with reduced feed conversion efficiency Urquhart et al. (1996); Spithill and Dalton et al. (1998). In tropical countries, fascioliasis is prevalent up to 90% and is considered the most important helminthes infection of cattle Spithill and Dalton et al. (1998). Most important, human can also become infected with Fasciola spp. and the disease is referred to as human fasciolosis. It is estimated that 2.4 million people in more than 60 countries are infected and the number of people at risk is more than 180 million throughout the world Haseeb et al. (2002); Mas-Coma et al. (1999); Ishii et al. (2002). Khalil et al. (2009) recorded 49.01% prevalence of fasciolosis infestation in buffaloes. He also observed that optimum infestation was seen from January to September, while minimum during October to December. Whereas Bhutto et al. (2002) recorded 4% buffalo calves infested with trematodes. Introduction 4 Morphological identification of Fasciola species requires significant parasitological skills and is not an accurate method of characterization, especially for the ‘intermediate’ form Kendall et al. (1965); Lin et al. (2007); Le et al. (2008). Hence, different molecular tools have been developed during the last decade for the accurate identification of Fasciola spp. Marcilla et al. (2002); Velusamy et al. (2004); Cucher et al. (2006); Magalhaes et al. (2008); Ai et al. (2010). Conventional techniques like fecal examination used for its diagnosis are not comprehensive because of Species similarity in distribution in many countries of East and North Africa, and Southeast and Central Asia, and are similar in egg morphology. Coprological methods continue to be the most widely used approach for the detection and quantification of Fasciola and other helminth eggs in laboratory animals, domestic and sylvatic animals and humans Cringoli et al. (2004, 2010); Bergquist et al. (2009), with a more recently developed method that is currently undergoing broad-scale validation the FLOTAC techniques Cringoli et al. (2010) for the detection and quantification of F. hepatica eggs in feces obtained from experimentally-infected rats before and after drug administration. FLOTAC has been used successfully in the diagnosis of F. hepatica infections in naturallyinfected sheep, which underwent treatment with standard Cringoli et al. (2006). However PCR based diagnosis using specie-specific primers has been proved more accurate and successful in diagnosis and distinction of Fasciola species Le TH et al. (2012). The molecular technique such as PCR is used for the recognition of parasite. These molecular techniques are largely accepted all over the world. These are more exact methods than traditional in epidemiological studies Heckeroth and Tenter et al. (1999). Herbal drugs have been used since ancient times to cure diseases and medicinal plants have been used to treat fasciolosis among these Nigella sativa and Allium sativum have been used to treat the worm infestation. No report of resistance development to plant anthelmintic although many have been used in veterinary medicine for many years Ashaal et al. (2010); Introduction 5 Waller et al. (1995); Koko et al.(2000); Thienpont et al. (1979); Keiser et al. (2011). Herbal (indigenous) drugs have been used since ancient times to cure diseases, and several medicinal plants have been used to treat fasciolosis Satyavati et al. (1987). Among these, Nigella sativa has been used to treat worm infestation Nadkrani et al. (1954; Said et al. (1969); Akhtar et al. (1988). N. sativa can be used as a therapeutic agent against helminthes Khan et al. (2013). Neem leaf powder can be used as therapeutic agent against gastrointestinal parasites of camel, equine, cattle, buffalo and small ruminants Mehmood et al. (2013). Garlic powder can also be used as herbal product against the gastrointestinal helminthes of livestock Ijaz et al. (2008).
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Agriculture sector plays a pivotal role in the economy of Pakistan. At present it
participates 11.4% to GDP. Agriculture provides 45% employment to labour of our country
and contributes in the development of other sectors of the economy. The livestock sector
occupies a distinguishing position in the National Agenda of economic development. It
provides net source of foreign income. In the history livestock has been dominated by small
holders to meet their needs of milk, food safety and cash earnings on daily basis. Besides,
livestock is considered a source of employment generation at rural level. It plays an important
role in poverty mitigation and keeps in elevating the socio-economic situation of our rural
commonalities. Livestock share approximately 55.4 percent to the agricultural value added
and 11.9 percent to national GDP during 2013-14. Anonymous (2013-14).
Parasitism is one of the major problems lowering livestock productivity round the
earth Vercruysse and Claerebout et al. (2001). The significance of helminthes infections is
increased manifold in developing countries like Pakistan where 65.2% population is rural
(Population Census Organization (2007) depending upon livestock for their incomes and
where parasitism acts as a serious trouble for livestock economy Chaudhry et al. (1984).
Among helminthes infections, fasciolosis commonly called as liver fluke disease is of vital
importance due to its wider spectrum of ultimate hosts Rondelaud et al. (2001) causing acute
and chronic infections Sampaio-Silva et al. (1996). The disease is primarily caused by
Fasciola hepatica and Fasciola gigantica Soulsby et al. (1987). Fasciolosis is a parasitic liver
infection of wild and domestic ruminants caused by genus Fasciola, which has worldwide
distribution Soulsby et al. (1986). Fasciolosis causes economic losses as a result of
mortalities, abortions, retard growth, reduced milk and meat production, condemnation of
infected liver and emaciated carcasses and cost of animal treatment Gracey and Collins et al.
(1992). Fasciolosis caused by F. hepatica is a cause of important economic losses in
ruminants. The disease is widely spread throughout in Africa, Asia and the Pacific. Although
mostly a problem of young stock, the disease is also found in older animals where it
adversely affects health, growth rate and development. Apart from its great veterinary
importance throughout the world, fasciolosis caused by both F. hepatica and F. gigantica has
recently been shown to be a re-emerging and wide spread zoonosis affecting a number of
human populations Mas-Coma and Bargues et al. (1997); Esteban et al. (2003).
Prevalence of fasciolosis infestation in buffaloes is 49.01%. The infestation of
fasciolosis was optimal from January to September, while least during October to December.
Fecal samples indicated F. hepatica prevalence 65%. Whereas prevalence of F. gigantica
was observed higher in adult cattle as compared to cattle calves by fecal egg counts Khalil et
al. (2009). 4% Buffalo calves infested with trematodes. Availability of intermediate host and
the grazing habits of the final host determines the epidemiology and seasonal pattern of
infection with trematodes Pfukenyi et al. (2005); Bhutto et al. (2002). The life cycle of F.
hepatica includes many stages which develop in the environment or the intermediate host.
Metacercariae which is the infective stage encysted and ingested by grazing herbivores.
Temperature and rainfall are the important factors affecting the life cycle and the prevalence
of F. hepatica infection McCann et al. (2010). Many studies revealed that F. hepatica is
endemic in buffaloes, cattle, goats, sheep and humans in Pakistan Ijaz et al. (2009); Iqbal et
al. (2007); Khan et al. (2010) and Qureshi et al. (2005).
Fasciolosis is the disease by which about 250 million sheep and 300 million cattle are
potentially affected worldwide Boray et al. (2005), and more significantly infects all
ruminants Iqbal et al. (2007). F. hepatica and F. gigantica are the two liver flukes commonly
reported to cause fasciolosis in ruminants Walker et al. (2008). Infected cattle can exhibit
Introduction
3
poor weight gain and dairy cattle have lower milk yield, and possibly metabolic diseases
Mason et al. (2004). Losses due to fasciolosis in the United Kingdom and Ireland alone are
more than £18 million a year Mulcahy and Dalton et al. (2001); €52 million a year or €299
per infected animal in Sweden Schweitzer et al. (2005); 0.26 million USD annual due to
fasciolosis associated liver condemnations in cattle slaughtered in Kenya Kithuka et al.
(2002). In Pakistan, 35697 USD (3141360 PKR) are reported in only one tehsil of Punjab
province, the Sargodha T.U. Rehman et al. (2013). While Fasciolosis is prevalent in whole
Punjab province of Pakistan but the marshy areas such as regions of Gujranwala, Lahore,
Multan, Faisalabad, Jhang, Muzaffargarh, Sheikhupura, and Sargodha are of major concern.
Fascioliasis in ruminants causes substantial economic losses, estimated at US$ 2
billion per annum worldwide, to rural agricultural communities and commercial animal
producers due to death of infected animals, condemnation of affected livers and production
losses associated with reduced feed conversion efficiency Urquhart et al. (1996); Spithill and
Dalton et al. (1998). In tropical countries, fascioliasis is prevalent up to 90% and is
considered the most important helminthes infection of cattle Spithill and Dalton et al. (1998).
Most important, human can also become infected with Fasciola spp. and the disease is
referred to as human fasciolosis. It is estimated that 2.4 million people in more than 60
countries are infected and the number of people at risk is more than 180 million throughout
the world Haseeb et al. (2002); Mas-Coma et al. (1999); Ishii et al. (2002). Khalil et al.
(2009) recorded 49.01% prevalence of fasciolosis infestation in buffaloes. He also observed
that optimum infestation was seen from January to September, while minimum during
October to December. Whereas Bhutto et al. (2002) recorded 4% buffalo calves infested with
trematodes.
Introduction
4
Morphological identification of Fasciola species requires significant parasitological
skills and is not an accurate method of characterization, especially for the ‘intermediate’ form
Kendall et al. (1965); Lin et al. (2007); Le et al. (2008). Hence, different molecular tools have
been developed during the last decade for the accurate identification of Fasciola spp.
Marcilla et al. (2002); Velusamy et al. (2004); Cucher et al. (2006); Magalhaes et al. (2008);
Ai et al. (2010). Conventional techniques like fecal examination used for its diagnosis are not
comprehensive because of Species similarity in distribution in many countries of East and
North Africa, and Southeast and Central Asia, and are similar in egg morphology.
Coprological methods continue to be the most widely used approach for the detection and
quantification of Fasciola and other helminth eggs in laboratory animals, domestic and
sylvatic animals and humans Cringoli et al. (2004, 2010); Bergquist et al. (2009), with a more
recently developed method that is currently undergoing broad-scale validation the FLOTAC
techniques Cringoli et al. (2010) for the detection and quantification of F. hepatica eggs in
feces obtained from experimentally-infected rats before and after drug administration.
FLOTAC has been used successfully in the diagnosis of F. hepatica infections in naturallyinfected
sheep, which underwent treatment with standard Cringoli et al. (2006). However
PCR based diagnosis using specie-specific primers has been proved more accurate and
successful in diagnosis and distinction of Fasciola species Le TH et al. (2012). The molecular
technique such as PCR is used for the recognition of parasite. These molecular techniques are
largely accepted all over the world. These are more exact methods than traditional in
epidemiological studies Heckeroth and Tenter et al. (1999).
Herbal drugs have been used since ancient times to cure diseases and medicinal plants
have been used to treat fasciolosis among these Nigella sativa and Allium sativum have been
used to treat the worm infestation. No report of resistance development to plant anthelmintic
although many have been used in veterinary medicine for many years Ashaal et al. (2010);
Introduction
5
Waller et al. (1995); Koko et al.(2000); Thienpont et al. (1979); Keiser et al. (2011). Herbal
(indigenous) drugs have been used since ancient times to cure diseases, and several medicinal
plants have been used to treat fasciolosis Satyavati et al. (1987). Among these, Nigella sativa
has been used to treat worm infestation Nadkrani et al. (1954; Said et al. (1969); Akhtar et al.
(1988). N. sativa can be used as a therapeutic agent against helminthes Khan et al. (2013).
Neem leaf powder can be used as therapeutic agent against gastrointestinal parasites of
camel, equine, cattle, buffalo and small ruminants Mehmood et al. (2013). Garlic powder can
also be used as herbal product against the gastrointestinal helminthes of livestock Ijaz et al.
(2008).

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