doi:10.5455/vetworld.2013.433-439 Prevalence of gastrointestinal parasites and Cryptosporidium species in extensively managed pigs in Mekelle and urban areas of southern zone

Aim: The objective of this study was to investigate prevalence of gastrointestinal parasites and Cryptosporidium species in extensively managed pigs in Mekelle and urban areas of southern zone of Tigray Region, Ethiopia during June - September, 2012. 
 
Material and methods: Seven hundred fourteen pigs of different ages and sexes were selected for fecal sample collection. Fecal samples were collected from the rectum of pigs with strict sanitation. A total of 25 soil samples were also collected from backyards of pig pens using clean zipped plastic bags. Both fecal and soil samples were examined for eggs and cysts of GIT parasites by flotation and sedimentation techniques. Modified Ziehl – Neelsen technique was used to examine oocysts of Cryptosporidium species from 276 randomly selected fecal samples. 
 
Results: Out of 714 pigs examined through flotation and sedimentation, 27.3% were infected by at least one gastrointestinal parasite. Ascaris suum (25.9%) was the most prevalent parasite followed by Fasciola hepatica (1.8%), Eimeria spp. (1.7%) and Trichuris suis (0.3%). There was no significant association between sex and prevalence of parasites ÷2[df 1] = 1.921; P=0.166). Contrary to this, age of pigs had effect on prevalence of parasites ÷2[df 2] = 8.376; P=0.015). About 7% of pigs examined were positive for oocysts of Cryptosporidium spp. Moreover, 72% of the soil samples found to be contaminated with eggs of Ascaris spp. in the study area. Apart from causing morbidity in the infected pigs, the potential of Ascaris of pigs to infect man and vice versa together with poor environmental hygiene, may complicate the epidemiology and control of Ascariasis in the study areas. Extensively managed pigs may also act as potential reservoirs for zoonoses of Cryptosporidium species. 
 
Conclusion: It is concluded that further investigations are crucial on molecular characterization of Ascaris and Cryptosporidium isolates of extensively managed pigs to determine the parasites upto species level so as to suggest proper control strategies.


Introduction infection of pigs with GIT parasites [1]. Gastrointestinal helminths including Oesophagastomum dentatum, Infection of pigs with gastrointestinal (GIT)
Trichuris suis, Ascaris suum, Oesophagastomum parasites is widely reported from all corners of the quadrispinulatum, Trichostrongylus axei, Strongyloides world and shown to be influenced by the type of pig ransomi, Hyostrongylus rubidus and Physocephalus management practiced [1].Extensive production of sexalutus have been identified in pigs raised under pigs for commercial purpose is widely practiced in extensive production in Kenya, Ghana and Burkina developing countries of Africa.Availability of cost free Faso [3-5].Semi-extensively managed pigs are feed (household or municipal garbage) and reported to harbor Taenia solium, in Nigeria [6].Pigs possibilities for the animals to get better nutrition rose under semi-intensive and extensive husbandry were through scavenging initiate pig growers of developing found to be infected with Ascaris suum and Oesophacountries to prefer extensive management.In this gostomum spp. in Holeta, Ethiopia [7].In addition to regard 70-95% of pig raising sectors of developing helminths, extensively managed pigs are also reported countries practice extensive husbandry [2].
to harbor intestinal protozoans including Cryptosporidium Poor environmental hygiene coupled with spp., Giardia lambelia, Balantidium coli and Eimeria extensive managements is reported as risk factors of spp. in developing countries [8].Among other intestinal protozoans Cryptosporidium has been given much attention because of its occurrence in various domestic and wild animals and and its potential for causing emerging cryptosporidiosis [9].average 600 mm of rainfall annually and more than have been identified [9] however, over 60 genotypes 70% of it falls between July and August, followed by remain undefined [10].Recent genetic characterization long dry season (October to May) [16].Mekelle has an studies revealed that pigs are infected with genetically estimated total human population of 215,546 [17].distinct and host-adapted form of Cryptosporidium, Urban agriculture is a common practice in Mekelle Cryptosporidium "pig"genotype [11].However, pigs [18].The livestock management system practiced in can also be infected with the zoonotic Cryptosporidium Mekelle includes extensive, intensive and semiparvum, "cattle" genotype, indicating their potential intensive [19].Pigs are recently introduced into as reservoirs of zoonotic infection to humans [11].

Prevalence of gastrointestinal parasites and
Mekelle and raised by few households under extensive So far Haileyesus [12], reported prevalence of management.Cryptosporidium parvum and C. hominis in extensively managed cattle and HIV/AIDS patients in Ethiopia.However, there is lack of data regarding the prevalence of Cryptosporidium spp. in extensively managed pigs in Ethiopia.Hence, it is crucial to determine the prevalence of Cryptosporidium species in pigs before classifying them as one of the reservoirs for zoonotic cryptosporidiosis in Ethiopia.
Pig infected with GIT parasites had poor feed conversion rate and delays in achievement of market weight.Some of the GIT parasites of pigs result in condemnation of organs or entire carcasses causing economic losses in pork industry [13].In addition, pigs infected with GIT parasites may act as source of zoonoses through contaminating the environment with infective stages of intestinal parasites present in their excreta [11].The epidemiology of pig born GIT parasites may further be complicated when some of the parasites of pigs infect man and vice versa.Ascaris spp. is typical example [14].its infancy in the country with a total population of northern Ethiopia (longitude 39 5'-39 8' and latitude 0 0 about 1900 [15].Recently small scale pig husbandry is 12 3'-13 7' (Fig- 1).Here agriculture involves mixed being practiced in urban areas of Ethiopia, however, crop and livestock farming.The climate is the management is mainly extensive whereby pigs are characterized by bimodal rainfall.About 70-80% of allowed to scavenge on household wastes at backyards the rain falls in the Kiremt season (June-September) and municipal garbage dumping sites at peri-urban [20].The mean minimum and maximum temperature 0 0 areas [7].Such extensive pig husbandry coupled with ranges from 8 C to 30 C, with small annual variations.poor environmental hygiene and pigs' voracious This zone is composed of 11 districts (Weredas).feeding behavior may render infection of the animals According to the 2007 census, the total urban and rural with helminth and GIT parasites in Ethiopia.human population of the zone is estimated to be Nevertheless, few studies have been done on 1,004,558 [17].This zone includes major urban areas prevalence of GIT parasites in extensively managed of Alamata, Mekoni, Korem, Maychew and Adigudom.pigs in Ethiopia in general and Tigray Region in Recently the number of households engaged in extensive particular [7].Therefore, this study was initiated to pig husbandry are increasing in urban areas of southern investigate prevalence of GIT parasites in extensively zone of Tigray region.managed pigs and associated environmental health Animals studied: This study was conducted during risks in Mekelle and urban areas of southern zone of June -September 2012.Seven hundred fourteen Tigray Region, Ethiopia.poured through a tea strainer into a beaker.Then owned by a total of 38 households in Mekelle and urban suspension was poured from the beaker into a test tube areas of southern zone of Tigray Region were included leaving a convex meniscus at the top.A cover slip was in the study.On the basis of the pig owning household then carefully placed on top of the test tube and the test head's declaration, the animals were categorized into tube was let to stand for 20 minutes and after that cover three age groups of < 5 months old, 5-12 month old and slip was carefully lifted from the tube together with a > 12 months old.Distribution of the pigs with respect to drop of fluid adhering to it and immediately placed on a sex and age is given in Table-1.
glass slide for microscopic examination [21].These procedures were used to recover eggs intestinal Collection of fecal and environmental samples: Fecal helminths and oocysts of Eimeria spp.samples were collected directly from the rectum of pigs with strict sanitation in the morning, when households Sedimentation: In case of using NaCl as a flotation solution, the flotation technique was supplemented release the animals from their pens.About 5 gram of with sedimentation to recover eggs of intestinal the collected fecal sample was put in screw cap bottles helminths such as Fasciola hepatica which do not float containing 10% formalin.Soil samples were collected well in sodium chloride solution.For this purpose, 3 g from backyards of pig pens using clean zipped plastic of fecal or soil samples was put in a conical flask and bags.Fecal and soil samples were transported to the mixed with 30 ml of water; the mixture was then sieved Parasitology Laboratory, Department of Biology, through a tea strainer into a beaker and transferred into Mekelle University (Ethiopia) on same day and stored a centrifuge tube for centrifuging at 1500 rotation per at -20 °C until processed and examined for eggs and minute for 3 minutes.The supernatant was then oocysts of gastrointestinal helminths and protozoans, discarded and sediment was mixed with 1% of respectively.
methylene blue and examined under the microscope Processing and examination fecal and soil samples using 40objective lens [22].Zinc sulphate flotation: Three grams of fecal samples were mixed with 50 ml of 0.4g/ml Zinc Sulphate Modified Ziehl-Neelsen technique: A total of 276 (flotation fluid) using mortar and pistil.Similarly soil randomly selected pig fecal samples from Mekelle and samples were also mixed with 50 ml of 0.4g/ml Zinc urban areas of southern zone of Tigray Region were Sulphate.The resulting fecal or soil suspension was examined for Cryptosporidium oocyts by the modified  Ziehl -Neelsen technique [21].Thin fecal smear was households.Fifty percent of the pig owning houseprepared from each of the fecal samples, air-dried, holds dump slurry of the animals in the nearby agricultural fixed with absolute methanol and stained with a carbol-farms as manure (Table -2).The mean number of family fuchsin (primary stain) for 30 minutes.The smear was size and pigs owned by households were found to be then washed with tap water and decolorized with 1% 4.05+0.379and 20.82+2.89,respectively.acid-alcohol (1ml HCl and 99ml of 96% ethanol) for 2 Out of 714 coprological samples of pigs minutes; washed again with tap water and examined, 195 (27.3%) were positive for at least one of the gastrointestinal parasites in Mekelle and southern counterstained with 1% methylene blue for another 2 zones of Tigray Region (Table -3).During microscopic minutes, then rinsed again in tap water and air-dried.
On the other hand, 3 (1.69%)pigs were found to

Survey for demographic information and pig husbandry
be positive for oocysts of Eimeria spp.(Fig. 3, Table 4).conditions: A close ended interview questionnaire About 26% of the parasitized pigs were infected with containing variables including characteristics of pig A. suum either as a single or multiple infections with owning households and demographic information was other parasite species.However, there is no statistically first prepared in English and then translated into the significant association in the prevalence of intestinal local language of Tigriyna, to collect data on pig parasites in pigs between Mekelle zone and southern owning households.Interviews were administered to a 2 zone of Tigray Region (χ [df 1] = 0.320; P=0.572).

total of 38 household heads encountered during the
There is also no association in the prevalence of study.Observational check list containing variables parasites between male and female pigs in the study such as presence of toilet and tap water supply for the 2 areas (χ [df 1] = 1.921;P=0.166).On contrary, there is household, site for dumping of pig slurry and floor significant association between different age condition of pig pens was prepared to collect data on categories and prevalence of parasites, where majority pig husbandry condition at household level.
of infection occurring in pigs with ages ranging from 5-2 Statistical analysis: Descriptive statistics was used to 12 months (χ [df 2] = 8.376; P=0.015).summarize data on prevalence of gastrointestinal Moreover, Out of 276 fecal samples examined for parasites in pigs and contamination of soil by helminth oocytss of Cryptosporidium spp.(Fig. 4), 20 (7.2%) eggs in Mekelle and urban areas of southern zone of were found to be positive.Tigray Region, Ethiopia.Chi-square test using SPSS10 Regarding parasitological contamination of the [23] software was conducted to see if there is any environment in the study areas, soil samples collected difference between prevalence of gastrointestinal from backyards of pig pens were found to be parasites with respect to age and sex of the pigs as well contaminated with eggs of Ascaris spp.(Table -5).as among different study sites.

Discussion Results
In this study it was possible to recover eggs and A total of 38 male household heads with ages oocysts of gastrointestinal helminths and protozoans, ranging from 15-69 years were identified engaged in respectively.Twenty seven percent of the pigs were extensive pig management in Mekelle and urban areas infected with one or more gastrointestinal parasite of southern zone of Tigray Region.Pig husbandry was species.Ascaris suum was the most common helminth a fulltime occupation for about 22 (58%) of the in all age categories of pigs examined in the present study.This is in agreement with similar studies in different districts of the region for the Abregele meat extensively managed pigs [24,25].On the contrary, factory.Irrigated urban and pre-urban crop production Tiwari et al. [26] reported no evidence of A. suum practice through ditching urban wastewater may render infection in pigs in Grenada, West Indies.Though, A.
suitable habitats for snail intermediate hosts of F suum is naturally a parasite of pigs, but it can also infect hepatica in the present study areas.From our personal human [14].The potential of A. suum to infect human observation, pigs in the study areas browse grasses might be due to the fact that it shares similar protein along the irrigation water ditches which may expose molecules with Ascaris lumbricoides for which man is them to infection with the metacercaria of F hepatica the natural host [27,28].Therefore, it is not surprising [33].This may also contributes for contamination of to say extensively managed pigs in the present study the ditched water with eggs or oocyts of gastroareas may act as potential reservoir hosts of human intestinal parasites contained in pig feces.ascariasis.
The prevalence of Eimeria spp. in this study was The prevalence of the second nematode parasite, much lower than a similar survey in Ethiopia [7]   municipal garbage in both Mekelle and urban areas of managed pigs in Mekelle and southern zone of Tigray southern zone of Tigray Region.Even in few sites of Region, Ethiopia.A. suum was most prevalent either as the study areas it was recorded that children were a single or multiple infections with other parasites in defecating outside to their home and pigs were allowed pigs examined.The possibilities of Ascaris spp. of pigs to feed on children's fecal material.Consumption of to infect man and vice versa coupled with unhygienic human fecal material by pigs may complicate the environmental condition may complicate the epidemiology of gastrointestinal parasites such as epidemiology and control of ascariasis in the study Ascaris suum and Cryptosporidium spp. that can infect areas.In the present study extensively managed pigs both pigs and man [10].
are also reported to harbor Fasciola hepatica and There was also no statistically significant Cryptosporidium spp.suggesting the potentials of pigs difference in the prevalence rate of gastrointestinal as reservoirs hosts of the parasites.Further investigations 2 parasites between male and female pigs (χ [df 1] = are recommended for molecular characterization of 1.921; P=0.166).This result is consistent with the Ascaris and Cryptosporidium isolates of pigs to clearly finding of previous study in Holeta, Ethiopia [7].
determine the species of the parasites responsible for However, Tamboura et al. [4] reported higher swine ascariasis and cryptosporidiosis, respectively in prevalence of gastrointestinal nematodes in female the study areas.pigs than in males in Burkina Faso.The reason for

Fig. 1 :
Fig.1: Map of the study area In Ethiopia the population of pigs has shown slight increment since 1980.However, pig raising is at Southern zone of Tigray Region is located in 0 0

Fig. 2 :
Fig.2: Eggs of gastrointestinal helminths isolated from extensively managed pigs in Mekelle and urban areas of Southern Zone of Tigray Region, Ethiopia; Ascaris suum (a), Fasciola hepatica (b) and Trichuris suis (c)

Fig. 3 :Fig. 4 :
Fig.3: Oocyst of Eimeria species isolated from extensively managed pigs in Mekelle and urban areas of Southern Zone of Tigray Region, Ethiopia Fig.4: Oocysts of Cryptosporidium species detected from extensively managed pigs in Mekelle and urban areas of Southern Zone of Tigray Region, Ethiopia.

Cryptosporidium species in extensively managed pigs in Mekelle and urban areas of southern zone of Tigray region, Northern Ethiopia
This study was carried out in Mekelle and that causes severe diarrhea, anorexia and weight loss, Southern Zones of Tigray Region, Ethiopia (Fig-1).especially in immunocompromised animals and humans Mekelle is the capital city of Tigray Region located 780 (mainly children bellow five years of age and HIV/ km north of Addis Ababa, the capital city of Ethiopia.AIDS patients).Cryptosporidiosis is caused by severalIts geographic location is 13° 32' N and 39° 33' E. It has genotypes and phenotypically diverse Cryptosporidium an average altitude of 2200 m.a.s.l with a mean species and has been emerging over the past decade.

Table - 1
. Distribution of sample pigs (n=714) with respect to sex and age in Mekelle and urban areas of Southern Zone of Tigray Region, Ethiopia

Table - 2
. Demographic information and pig husbandry conditions practiced by households in Mekelle and Southern Zone of Tigray Region, Ethiopia and Trichuris suis, was very low in different age groups of Zimbabwe [34], respectively.This difference in pigs examined.This low prevalence agrees with the prevalence of Eimeria spp.may be attributed to the findings of earlier studies in Africa and West Indies [26, underlining difference in epidemiology of Eimeria spp.29, 30].This supports the speculations that T. suis eggs in the areas and methods used to recover oocysts from are highly susceptible to environmental factors.The fecal samples.prevalence of Fasciola hepatica was low (2.81%) in In the present study, out of 276 pigs examined 20 this study.On the contrary, Apt et al. [31] reported (7.2%) were positive for Cryptosporidium spp.This higher prevalence F hepatica in pigs of the Chilean result is consistent with the findings of Morgan et al.This may be rural villages to markets in the urban areas may due to the reason that pigs of both of the study areas are contribute for the introduction of gastrointestinal under similar risk factors of infection with gastroparasites including F hepatica to the urban areas.In intestinal parasites.During our field survey it was addition, there is huge cattle transport to Mekelle from observed that pigs were scavenging on household and Prevalence of intestinal parasites in pigs in Mekelle and Southern Zone of Tigray Region, Ethiopia (2012) A= Ascaris suum, F= Fasciola hepatica, E= Eimeria spp., T= Trichuris suis; *Single infection means, infection of pigs with any one of the rd parasites; #For example, in 3 column it means 83 pigs were infected with Ascaris suum (A), 01 with Fasciola hepatica (F) and 01 with Eimeria spp.(E) and similarly for other rows in the same columns.@Multiple infections mean, pigs infected with more than one parasite. @Table-3.

Table - 4
. Prevalence of gastrointestinal parasites in different age groups of pigs in Mekelle and urban areas of Southern Zone of Tigray Region, Ethiopia* Table-5.Contamination of soil by eggs of Asacris spp. at backyards of pig pens in the study areas.