Albanian j. agric. sci. 2017; (Special edition)
Agricultural University of Tirana
(Open Access)
RESEARCH ARTICLE
Larval Ascaris suum migration and diagnostic value in pigs YLLKA (MIJA) ÇANI1*, BEJO BIZHGA2 1
PhD Candidate, Faculty of Veterinary Medicine, Agricultural University, Tirana, Albania
2
Faculty of Veterinary Medicine, Agricultural University, Tirana, Albania
*Corresponding author; E-mail:
[email protected]
Abstract Study focused on diagnostic alternatives of Ascaris suum infection in 162 pigs in slaughterhouse. The range and infestation intensities of Ascaris suum infection depend on age, period and method of examination. Coproscopic examinations from 162 samples showed 124 of them (76.54%) positive for Ascaris suum eggs. Coproscopic examinations resulted the most efficient and offer the possibility of epidemiological estimates. From the samples were found with injuries as a result of migration process 49 pigs or 30.24% of the sample surveyed. During the macroscopic examination of the intestine of pigs were found with the adult parasite in 78 heads or 48.14% of the examined pigs. This value was about 20% lower than the value of coproscopic examination, but about 20% higher than the value of the examination of the milk liver spots. During of the macroscopic and microscopic examination in lungs were found with signs of pneumonia and parasitic larva migration in the lung parenchyma and the bronchi, 58 heads or 35,8% of the examined pig lungs. In pigs when parasitic pneumonia detected, the nose leaks were examined for the presence and number of larvae. From 162 nose tampons examined resulted positive for the presence of Ascaris suum migration larvae 26 pigs (16.04%) from the total of examined heads. Post-mortem examinations in slaughterhouses at pigs resulted efficient, low cost and provide sufficient data for Ascaris suum infection. Keywords: Ascariasis; swine; method; larvae; diagnose.
1. Introduction
in their place is created the second stage larvae. At the temperature 20
0
C they become second-stage
Ascaris suum is a nematode that infects pigs and
larvae and they infest pigs when they eat food or
randomly it can affect people. A. suum is present
swallow water and invasive eggs, from which the
across in our country and causing major damage to the
small intestine out of inexpensive stage larvae II, to
growth economies of pigs and the disease causes
begin phased hepato-entero-pulmonary migration [10,
called ascariasis [4, 6, 13, 17]. Adult ascarids living in
12, 15, 17]. The stage II larvae pierce the intestines
the small intestine and the mature females produce
and enter the blood vessels (veins), beginning their
100,000-200,000 a day eggs that come along with the
migration to the liver, where they stay 4-5 days and
feces in the external environment. The male is 15-25
transformed into L 3 larvae that reach the liver
cm long and 0.3-0.4 cm wide, while the female is 25-
through the hepatic veins, pass in vena cava caudalis
41 cm long and 0.3-0.6 cm wide. It is a long worm,
and the right half of the heart, from which through the
the shaft-shaped yellow. Mouth are surrounded by
pulmonary artery into the lungs arrive 4-7 days after
three edge equipped with scarring little theeth. Eggs
infestation [3, 7, 14]. Pierce blood vessels and walls of
are 50-70x40-50μ, they have ellipsoidal shape, thin-
the alveoli, hit the alveoli and begin to climb in the
walled and small granules. They have brown to
airways, continuing movement and assisted by the
yellow colour, are non-segmented and very resistant.
mucociliary apparatus, the larvae emerge in the
Eggs maintain vitality in the land, but can be
pharynx, where together with saliva and bronchial
destroyed by the direct rays of the sun [4, 6, 8, 13,
fluids ingested and down to the intestine where it
27]. Eggs newly emerging with the feces are not
begins the second phase or intestinal stages. The
infestive, they can be transformed invasive eggs when
larvae reappear on the day of the casing 8 and 9 after
369
Çani et al., 2017
infestation there doing stripping III and transformed
migration of the ascarids larvae [6, 12, 16, 18, 22].
into L4 and around day 30 to make stripping IV and
To differentiate parasitic stains from stains caused by
transformed into L5. The entire cycle of pig organs
fungi and bacteria samples from the liver in the
lasts 60-70 days and the life of A. suum lasts about
laboratory were stained with Wright Gimsae and Ziehl
one year [1, 9, 21]. A. suum exists everywhere where
Neelsen [16]. In the same piglets were observed also
pigs grow and the level of infestation ranges routine
the
from 20-70%. Infestation can be caused at all ages,
pneumonia or suspected cases of migration of larvae
but more serious are age 1-5 months. More infested
in the lungs when the hemorrhagic lesions were noted
are the pigs who do not eat well, and when lacking
and eosinophils infiltration around alveoli (the larvae
hygiene is bigger [12, 23, 27]. The source of the
migrate to the bronchial tree) were noted hemorrhage,
disease are infested pigs. The greatest danger comes
edema and emphysema [3, 8, 13, 25].
from manure from stalls without leave and employees
resulting suspicious or affected, were sampled from
who can spread the eggs wherever they move [1, 4, 6].
damaged areas, or to be fully examined in the perfusion method [17, 25, 28].
The examinations were conducted in 162 pigs in the slaughterhouse. Scope of work during coproscopic examination was to determine the prevalence and parasite load of A.suum in pigs in the according
to
epidemiological
alignment criteria, sampling and evaluation [2, 4, 6, 17, 23]. Fecal samples were taken directly to the right intestine which often were completely campionated. In this case the right hose broke away from the rest connected on both sides and the plastic bag was transported
in
laboratory.
For
coprologic
examinations were used qualitative and quantitative methods of sedimentation and flotation [4, 6, 17]. Examinations post mortun to the peculiarities of biology A.suum highlighted changes in the lungs and the liver and damages during the phase of hepatopulmonary migration of slaughtered pigs in the slaughterhouse. These surveys were carried out in these organs which often were sampled completely. We routinely by these organs were sampled and samples
for
macroscopic,
Lungs
laboratory. In lab full lungs were examined with
2. Material and Methods
slaughterhouse
lungs. After an observation of parasitic
histological
and
As for the debris
became microscopic examination of smears prepared by the
fluid in the bronchial
tree. At
the
slaughterhouse was prepared swabs from the nose leaks from all piglets that were used in the experiment [3, 14, 19].
Tampons were observed to find
migratory larvae. In cases when larvae were found calculates their number. To observe, intestines were divided into several parts by ligature and were cut into pieces 1-2 meters long and trasported introduced in plastic boxes. For examination of the intestine parts were washed in pure water and the water was collected and was surveyed after sedimentation [2, 7, 21].
After each rinse portion of the intestine was
opened by enteretom was surveyed and the interior to discover fixed worms mucus. To be separated from the mucous membrane do not retreat with pliers but torn mucous fixed around the country to favor secession. Content and mixed water filtered and the remaining filter disposed in a container to be examined [7, 9]. In the lab was raided all bowels and sediment rinse was tested on a glass tray located on a black sfod. For detection of small nematode was used
microscopic examination [11, 12, 15].
artificial digestion and accomplished overnight in 2.1. Procedures at slaughter
and post
thermostat at 37°C. After that was observed with low
mortum examinations
magnification dissolved material from which were
In slaughterhouses in livers of piglets was conducted
separated parasites [6, 17, 21, 26]. During micro and
evaluation of the milk spots. Livers were carefully
stereomicroscopic
observed for the evaluation of hot spots due to
collected, differentiated and counted young and adult
370
examination
was
observed,
Larval Ascaris suum migration and diagnostic value in pigs
parasites in the intestines of examined piglets [1, 3, 5,
Coproscopic examinations were conducted
24, 28]. Coproscopic examinations, the nasal and the
with alterations of the method of sedimentation and
majority of the organs were performed in the
flotation (Mc Master). In 72 (44%) stoll samples from
laboratory of veterinary parasitology, FVM.
the intestine (rectum and right intestine) were applied to both quantitative methods for the same sample [2,
3. Results and Discussion
4, 17, 23].
Table 1. Results of post-mortem examinations of pigs in the slaughterhouse.
No
Examination method
Sample piglets no
Positive no
Positive %
1
Coprology
162
124
76.54
2
Milk spots
162
49
30.24
3
Larval pulmone migration
162
58
35.8
4
Nasal swabs
162
26
16.04
5
Ascaids in intestine
162
78
48.14
Table 2. Parasitic loud under quantitative coproscopic examinations.
Examination method Parasitic loud
Sample piglets no 162
Average e/g/f 168
Variations e/g/f 10-1800
From coproscopic examinations were found
injuries were were noted in about one third of the pigs
the highest values of the infestation of Ascaris suum
examined. Liver examination for "milk spots" is valid
in piglets. The data confirm that coproscopic study
and provides enough information for the particular
regardless of the method applied is most successful
observations may be made in the slaughterhouse [4, 6,
methods for diagnosing ascariasis in piglets. It is fast,
12, 16, 26]. Examination is easy and free of cost and
easy and low cost. From 162 samples examined
status is sufficient to approvement conclusions for the
(coproscopy) 124 of them (76.54%) resulted positive
ascarids pathology in piglets. The opposite applies to
for Ascaris suum infection [4, 6].
the
From
coproscopic
observations
method
negativity.
Negativity
should
be
average
reconsidered and evaluated carefully, as not all
parasitic load in piglets resulted in 168 e/g/f with 10-
affected piglets may have the presence of milk spots
1800 e/g/f variations. During post-mortem inspection
in the liver. The data from the study showed that only
knowing the peculiarities of A. suum biology,
about a third of pigs affected by milk spots in their
examinations were carried out in the liver, lungs, nose
liver. These milk spots are classified in three models
and intestines. These surveys were carried out in
based on the macroscopic appearance. Histologically
slaughterhouses and by the same piglets that served as
the following three types of lesions are seen:
the basis of this study. From samples, were found with
interstitial hepatitis and eosinophilic intralobular
injuries as a result of liver migration 49 piglets or
necrosis, arteriolar degeneration and granuloma, and
30.24% of the observed samples. These injuries are
lymphofolicular
known as "milk spots" in the liver. This was the main
relationship between macroscopic and histological
characteristic of a chronic hepatitis caused by parasitic
patterns,
Ascaris suum larvae migrans. The spots are the result
manufactured by interstitial eosinophilic hepatitis [6,
of larval migration within the liver. These migratory
12, 16, 17]. With the naked eye, microscopic
371
compact
hyperplasia. milk
spots
Regarding are
the
generally
Çani et al., 2017
observations and histopathology were observed milk
This is followed by an intensive infiltration of
spots in the livers of pigs in the slaughterhouse. Livers
eosinophils. These lesions are visible at necropsy on
of piglets positive for milk spots were brought to the
the lung surface as white areas and are clearly
laboratory for thorough examination. Liver samples
separated from the healthy part of the body. In these
resulting with milk spots belonging to all ages of pigs
cases were found migrating larvae of A. suum [6, 12,
2-7 months. Lesions were in diameter between 0.5 and
19]. In piglets from leaks nose in 162 piglets were
1 cm distributed in whole liver parenchyma and
taken tampons in their noses and were prepared to
infiltration in hepatic parenchyma. Often larvae were
microscopic
noted in the parenchyma, or migrate freely. In cases of
observed in stereomicroscopes and microscopes as
chronic forms of larvae resulted in dead and blocked
wet and dry preparations. The technique proved to be
abscesses or granulomas. For differentiated parasitic
very present when it is known that the larvae appear
stains, or stains caused by fungi and bacteria samples
on the nose of the pig infested on day 7 and 9 after
from the liver were stained with Wright Gimsae and
infestation. Larvae in the nose and mouth swallowed
Ziehl Neelsen [12, 16].
During entero-hepato-
into the gut or leaving by sneezing or runny nose in
cardio-pnemo-intestinal migration in tissues where
the external environment. All samples proved positive
larvae pass, causing bleeding and increased volume of
for migratory larvae of A. suum. Swabs diagnostic
organs and necrotic spots. Lung larvae are reared and
technique proved to be very simple, extremely
cause multiple hemorrhages in the alveoli, the
efficient and the very fruitful outcome [14, 19, 23].
brokiola, east of edema and parasitic pneumonia in
Work is underway to convert into quantitative
pigs [3, 6, 12, 19]. The larvae produce toxins that act
technique for estimating the number of larvae and
and other organs of piglets. Lung larvae are reared and
other parasitic estimates.
cause
alveoli,
tampons tested positive for the presence of larvae 19
bronchioles, east of edema, formation of parasitic
piglets or 15.83% of the total examined tampons. This
pneumonia which is present in piglets. From samples
is the lowest value recorded for the same piglets
examined were found to damage as a result of
category. The reason is the method fragility and
migration in the lungs 58 piglets or 35.8% of the
biological features of Ascaris suum which only for a
observed samples. These migratory lung damage
short period of time can be found in the nasal leakage.
resulted in about 5% higher than milk spots that were
This is the period when he comes in after the
noted in about one third of the pigs examined. Lung
migration bronchioles and mucociliary apparatus
examination for the presence of migratory pneumonia
comes in the mouth to swallow or nose to be removed
resulted more efficient examination of the liver and
with leaks in the external environment [6, 19]. This
provide
in
time is limited and therefore values and will be
slaughterhouses [6, 12, 19, 21, 22, 26]. The larvae
smaller. The advantage of the method is that the
migration to the lungs are capable of causing an
examination method may bring valuable data and the
inflammatory reaction, destruction of pulmonary
alive piglets (not necessarily killed) and status and
tissue and parasitic pneumonia with hemorrhagic foci.
parasitic loads are more value [6].
multiple
hemorrhages
sufficient
data
for
in
the
observations
smears.
Microscopic
swabs
were
From 162 examined
Table 3. Parasitic loud under quantitative nasal tampon examinations.
Examination method
Samples no
Average l/ml
Variations l/ml
Nasal swabs parasitic loud
162
2.5
1-12
In our case average parasitic loud was 2.5 larva per ml found that the method was converted into 372
quantitative method and the number of larvae per ml ranged in values 1-12 l/ml.
In these piglets was
Çani et al., 2017
performed and intestinal observation to find and count
monitoring of post-mortem swine ascaridiasis. While
increased ascarids. While mature parasites were found
nasal examinations in young piglets can be used as an
in the intestines of 78 piglets or 48.14% of the
alternative method for the diagnosis and monitoring of
observed samples [6, 13].
ascaridiasis.
To resist intestinal
descending flow they stare ends of their body in the intestinal mucosa and the damage it mechanically. When numerous they block the movement of food and cause measures extensions, twisting, gut cracks and east of peritonitis. They can climb in the stomach and cause nausea. They absorb large amounts of protein and food stuffs [1, 6, 12, 19, 21, 23, 28]. 4. Conclusions Coproscopy resulted in successful methods for diagnosing ascaridiasis in piglets. It is the fast, easy and low cost. Of 162 samples examined 124 of them (76.54%) resulted positive for Ascaris suum eggs. From coproscopic observations average parasitic load in piglets resulted in 168 e/g/f with 10-1800 e/g/f variations. The migratory damage (milk spot) was seen in about 1/3 of the liver of the piglets examined, while in coproscopy were identified 76.5% of the samples taken in the same piglets. The difference is explained by the features of the biological cycle of the parasite and the specificity of the immune piglets. Lung examination for the presence of migratory pneumonia resulted in the efficient examination of the liver and provide sufficient data for observations in slaughterhouses. From samples examined were found to damage as a result of migration in the lungs 58 piglets or 35.8% of the observed samples. These migratory lung damage resulted in about 5% higher than milk spots. From 162 examined tampons tested positive for the presence of larvae 19 piglets or 15.83% of the total examined tampons. This is the lowest value recorded for the same piglets category. The reason is the fragility of method and biological features of Ascaris suum. While were found positive for askarids in the small intestine about 50% of the examinated piglets. Control of liver and lung where they are visible signs of pneumonia migration and the presence of invasive larvae, can be applied as successful diagnostic techniques for the diagnosis and 373
6. References
1. Anderson TJ: The dangers of using single locus markers in parasite epidemiology: Ascaris as a case study. Trends in parasitology 2001, 17, 183–188. 2. Arizono N, Yoshimura Y, Tohzaka N, Yamada M, Tegoshi T: Ascariasis in Japan: is pig-derived Ascaris infecting humans? Japanese journal of infectious diseases 2010, 63, 447–448. 3. Bernardo T. M, Dohoo I. R, Donald A: Effect of ascariasis and respiratory diseases on growth rates in swine. Canadian Journal of Veterinary Research 1990, 54, 278–284. 4. Bizhga B.: Parazitologjia veterinare. Pg. 391-419. Gear Tirane, 2013. 5. Boes J, Coates S, Medley G. F, Varady M, Eriksen L, Roepstorff A, Nansen P: Exposure of sows to Ascaris suum influences worm burden distributions in experimentally infected suckling piglets. Veterinary Parasitology 1999, 119, 509–520. 6. Çani M.Y, Bizhga B: Ascaris suum Infection Estimate. Anglisticum Journal (AJ), 2016, 5 (9), 8-13. 7. Carstensen L, Vaarst M, Roepstorff A: Helminth infections in Danish organic swine herds. Veterinary Parasitology 2002, 106, 253–264. 8. Coates S.: Modelling the population dynamics of Ascaris suum in pigs. PhD Dissertation. Danish Centre for Experimental Parasitology, Copenhagen, Denmark and the University of Warwick, Coventry, UK. 2000. 9. Criscione CD, Anderson JD, Sudimack D, Peng W, Jha B: Disentangling hybridization and host colonization in parasitic roundworms of humans and pigs. Proc Biol Sci 2007, 274, 2669–2677. 10. Crompton DW: Ascaris and ascariasis. Adv Parasitol 2001, 48, 285–375.
Larval Ascaris suum migration and diagnostic value in pigs
11. Dold C, Holland CV: Investigating the underlying mechanism of resistance to Ascaris infection. Microbes and infection/Institut Pasteur 2011, 13, 624–631. 12. Geenen P L, Bresciani J, Boes J, Pedersen A, Eriksen L, Fagerholm H.P, Nansen P: The morphogenesis of Ascaris suum to the infective third-stage larvae within the egg. Journal of Parasitology 1999, 85, 616–622. 13. Holland CV: Predisposition to ascariasis: patterns, mechanisms and implications. Parasitology 2009, 136, 1537–1547. 14. Jolie R, Backstrom L, Pinckney R, Olson L: Ascarid infection and respiratory health in feeder pigs raised on pasture or in confinement. Swine Health and Production 1998, 6, 115–120. 15. Jungersen G: Immunity and immune responses to Ascaris suum in pigs. In The Geohelminths: Ascaris, Trichuris and Hookworm (ed. Holland, C. V. and Kennedy, M. W.): Kluwer Academic Publishers. Boston, Dordrecht, London; 2002: 105–124. 16. Kakihara D, Yoshimitsu K, Ishigami K, Irie H, Aibe H, Tajima T, Shinozaki K, Nishie A, Nakayama T, Hayashida K, Nakamuta M, Nawata H, Honda H: Liver lesions of visceral larva migrans due to Ascaris suum infection: CT findings. Abdom Imaging, 2004, 29, 598-602.
21. Napravnik J, Berka T: Cause and effect of ascariosis in pig herds. Helminthologia 2002, 39, 171-183. 22. Ngowi HA, Kassuku AA, Maeda GE, Boa ME, Willingham AL: A slaughter slab survey for extra-intestinal porchine helminth infections in northern Tanzania. Trop Anim Health Prod, 2004, 36, 335-340. 23. Roepstorff A, Eriksen L, Slotved HC, Nansen P: Experimental Ascaris suum infection in the pig: worm population kinetics following single inoculations with three doses of infective eggs. Parasitology 1997, 115 (4), 443–452. 24. Roepstorff A.: Ascaris suum in pigs: population biology and epidemiology. Doctorate thesis, the Royal Veterinary and Agricultural University, Copenhagen, Denmark. 2003. 25. Slotved H.C, Barnes E.H, Eriksen L, Roepstorff A, Nansen P, Bjorn H: Use of an agar-gel technique for large-scale application to recover Ascaris suum larvae from intestinal contents of pigs. Acta Veterinaria Scandinavica 1997, 38, 207–212.
17. Kassai T.: Veterinary Helminthology. Butterworth-Heinemann, Oxford, UK. 1999.
26. Wang T, Van Steendam K, Dhaenens M, Vlaminck J, Deforce D, Jex A.R, Gasser R.B, Geldhof P: Proteomic analysis of the excretory-secretory products from larval stages of Ascaris suum reveals high abundance of glycosyl hydrolases. PLoS Neglected Tropical Diseases 2013, 7(10):e2467.
18. Masure D, Vlaminck J, Wang T, Chiers K, Van den Broeck W, Vercruysse J, Geldhof P: A role for eosinophils in the intestinal immunity against infective Ascaris suum larvae. Plos Neglected Tropical Diseases 2013, 7(3), e2138.
27. Weng YB, Hu YJ, Li Y, Li BS, Lin RQ, Xie DH, Gasser RB, Zhu XQ: Survey of intestinal parasites in pigs from intensive farms in Guandong province, Peoples Republic of China. Veterinary Parasitology 2005, 127(3-4), 333-336.
19. Mejer H, Wendt S, Thomsen L. E, Roepstorff A, Hindsbo O: Nose-rings and transmission of helminth parasites in outdoor pigs. Acta Veterinaria Scandinavica 2000, 41, 153–165.
28. Zajac A.M, Comboy G. A.: Veterinary clinical parasitology. Blackwell publishing. State Avenue, Ames, USA. 2006.
20. Miquel N, Roepstorff A, Bailey M, Eriksen L: Host immune reactions and worm kinetics during the expulsion of Ascaris suum in pigs. Parasite Immunology 2005, 27, 79–88.
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