Diagnosis of Eimeria spp. in Capra ibex

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Diagnosis of Eimeria spp. in Capra ibex ( local meriz )


Capra ibex ( var Kurdistani ? ) occur throughout Kurdistan, especially in mountains and valleys where they are in sometimes domesticated for local purposes .The public name is Meriz goats which are found along the northern border region of Iraq. This breed is hardy and is mainly raised for its fine fibers (Alkass and Merkhan,2013). Meriz was studied previously for the presence of some parasites (Al- Nakshabandi and Al- Bayati , 2007) but not for coccidial infection.

The number of Eimeria spp.( coccidia parasites ) which were considered a parasite for domestic goats are variable and controversial, also depend upon the acceptance of validity of some species (levine and Iven ,1970) but may reach 17 species ((Levine and Ivens,1986; Levine,1985; Levine,1988; Soe and Pomroy ,1992; Soe and Pomroy , 1992 ).

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Goats coccidiosis have many epidemiological and economical aspects according to species’ diversity and seasonal appearance which reflexes the prevalence and distribution of the disease’s infection rate.

Eimeriosis of small ruminants are hosted specifically and their infection usually causes individual clinically appearance infection but mostly is chronic and undiagnosed. Economic losses may be related to high rate of mortality in kids, poor growth, persistence of environmental pollution with coccidian Oocysts and the cost of anticoccidial medicates.

Current paper aims to investigate the presence of Eimeriosis in local merize (Capra ibex ) for the first time in Duhok area and detailing their feature, as well as some epidemiological categories will be investigate.

Material and methods

1- Fecal samples of 77 heads of local merize ( 37 heads > 6 months and 40heads < 6 months ) were collected directly from the rectum , as apart from a cross sectional study for the diagnosis of infection with coccidia protozoal parasite ( Eimeria spp. ).

2- Samples weight were in about 10-20 g , collected individually and transfer to laboratory for successive investigation.

3- Direct smear ,flotation in saturated salt solution, and culturing with 2.5% ,5% Pot. dichromate ( wt / vol ) methods at the room temperature are used for the diagnosis of various Eimera spp.

4- Photomonogram , micrometer microscopy , and parasitology lab- wares are used.

4- Statistical analysis was done with ANOVA , Chi square and other descriptive statistic methods.

Results

Eimeria spp. in local merize ( Capra ibex ) were diagnosed by observation of undeveloped Oocysts and fully mature one that contains 4 sporocysts with various diagnosticmethods.

In a relation to the infection rate, kids ( > 6 months ) shows high infected rate ( 17 / 77 = 22.08%)

Compared to others that they are over 6 months age ( 12/77 = 15.59%) with total infection rate of about 29/77 ( 37.67%) as a cross sectional study results ( tab.1).

The relationship between infection and clinical signs onset only in 3 ( 3.9%) suffering from moderate diarrhea and only in small age (≤ 6 months ) animal while the rest had normal fecal formation( tab1). Sporulation time clear to be in around 7-10 days for allover species with 5% Pot. dichromate where 2.5% was not applicable for that ( tab2).

Six Eimeria spp. were recognized among infected animal with sporadic or mixed infection ( tab 2 ). The measurement ( in Micrometer - µm) of these species are differentiated according to dependant taxonomy kit (Eckert et al., 1995 ) 21 ), they are arranged from 16.03-15.03 (±0.55)µm for Oocyte, 0.42±0.09µm for Embryonic mass area to L x W ratio, Not clear germinal disk was observed in pre sporulation Oocyst , 4 sporocysts are present with 7x8 µ m in there measurement with no polar cup neither micropyle for the Oocyst , sporullation time was 7-10±2 days at room temperature . The suspected diagnosis is Eimeria( E.) alijevi ( fig. 1).

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The second suspected species was E. christenseni (fig. 2) with measurement of 24.04 -36.07 (±1.61) µ m for Oocyst , 0.18±0.08 µ m for Embryonic mass area to L x W ratio, clear germinal disk with clear Outer ( 1.6x2.08 ± 0.25 µm ) and Inner ( 0.83-1.25 ±0.25 µm ) layers was observed in pre sporulation Oocyst , 4 sporocysts are present with 7x11 µ m in there measurement , with polar cup and micropyle for the Oocyst . The sporullation time was 7-10±2 days at room temperature also.

The third suspected species was E. caprovina (fig. 3) with measurement of 26.45-31.26 (±2.41) µ m for Oocyst , 0.18±0.08 µ m for Embryonic mass area to L x W ratio, clear germinal disk and Clear Outer ( 1.6x2.08 ± 0.25 µm ) and Inner ( 0.83-1.25 ±0.25 µm ) layers was observed in pre sporulation Oocyst , 4 sporocysts are present with 8x10 µ m in there measurement ) with no polar cup but micropyle. The sporullation time was 7-10±2 days at room temperature also.

The fourth suspected species was E. minasensis (fig. 4- up ) with measurement of 28.86 -40.88 (±1.89) µ m for Oocyst , 0.18 ±0.09 µ m for Embryonic mass area to L x W ratio, clear germinal disk with clear Outer ( 1.6x2.08 ± 0.25 µm ) and Inner ( 0.83-1.25 ±0.25 µm ) layers was observed in pre sporulation Oocyst , sporocysts measurement were not follow up .There was no polar cup but micropyle clear in the Oocyst . The sporullation time was not detected.

The fifth suspected species was E.megaembryonica?? (fig. 4- mid ) with measurement of 33.67 -40.88 (±0.92) µ m for Oocyst, 0.67±0.07 µ m for Embryonic mass area to L x W ratio, clear germinal disk with clear Outer ( 1.6x2.08 ± 0.25 µm ) and Inner ( 0.83-1.25 ±0.25 µm ) layers was observed in pre sporulation Oocyst . Sporocysts measurement were not follow up .There was no polar cup but micropyle clear in the Oocyst. The sporullation time was not detected.

The sixth suspected species was E. ninakohlyakimovae (fig. 4 , down) with measurement of 14.34-19.24 (±1.44) µ m for Oocyst , 0.39±0.09 µ m for Embryonic mass area to L x W ratio, clear germinal disk with clear Outer ( 1.6x2.08 ± 0.25 µm ) and Inner ( 0.83-1.25 ±0.25 µm ) layers was observed in pre sporulation Oocyst . Sporocysts measurement were not follow up .There was no polar cup but micropyle clear in the Oocyst . The sporullation time was not detected.

In a comparison with diagnostic methods ( tab. 3 ), direct culturing with 5% Pot. Dichromate and saturated salt solution are more sensitive ( P=0.001 ) than direct wet smear preparation or culturing with 2.5 % Pot. Dichromate.

Discussion

The Infection rate of local meraz Capra ibex coccidiosis reach 37.6% which is less than what has been recorded in other goat species like that of Poland ((Balicka-Ramisz, Ramisz, Vovk, , and Snitynskyj,2012 ) 9) , Iran ((Radfar et al.,2011), Spain (Ruiz et al. , 2006 ) 11), Turkey (Deuer et al., 2003 ; Gul , 2007), Pakistan ( Rehman et al.,2011 ), Jordan (Mahmoud and Hossam,2003 ) and Czech (Strnadová et al.,2008 ), 17 ) as well as for adult (15.6% ) is lesser than others ( Mahmoud and Hossam,2003 ; Strnadová et al., 2008). These variation in infection rate may be related to host-parasite relationship as Capra ibex is a wild species and some time used as a semi domesticated animal, and also the hot climate of the area may prevent the higher infection rate from rising. This conditions were stated in previous studies ( Deuer et al.,2003 ; Gul , 2007 ).

In relation with clinical signs, diarrhea was the most observed sign in adequate with others ((Bakunzi, et al., 2010 ; Bakunzi et al. , 2010 ; Bandyopadhyay,2004.Sporulation time was 7-10 days which resembles the most of other studies (Soe and Pomroy,1992 ;Gul , 2007 ) and that may be related to essential temperature and humidity.

Six species of coccidia were diagnosed with current study according to available diagnosis key (Levine, 1985 ; Soe and Pomroy,1992 ; 11 Ruiz et al. , 2006 ; Deuer et al.,2003; Silva and Lima,1998; Eckert et al. 1995) .The infection was mixed in all cases but number of species in each case was not determined. These mixed infection occurrence may be attributed to free feeding style of Capra ibex which was dependent on the geographical area of this research. The mixed infection has been observed mostly in previous studies ((Deuer et al.,2003; Gul , 2007;Strnadová et al.,2008;Chhabra and Pandey,1991 ; Deger et al. , 2003) which were dealing with the same subject. A suggestion of new species in Capra goat should be expected and putting in mind, because of nature of the studied animal and the measures of the Oocyst .The suggested species name was E.megaembryonica?? (fig. 4- mid ) because of higher (0.67±0.07 µ m) Embryonic mass area to L x W ratio. This species needs more conformational studies .

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Also the relationship between Oocyst total size and embryonic mass could be used in classification of different species and that what was observed in the current study for the first time. This subject depend on two dimensional measurements as by multiplying length and wide of a particular Oocyst and the circular area rule for the embryonic mass area. Other methods related to three dimensions technology may be more conventional as stereoscopy facilities.

Other criteria is the observation of germinal disk which composed from two parts outer and inner. The outer was clear and transparent while the inner was gray and condense with fixed dimensions in allover detected species. This foundation mentioned for the first time for coccidia and needs more investigation with advanced laboratory techniques like Electron microscopy.

Lately , it should be mention that current study showed that 5% Pot. dichromate is better for clear sporulation in compare with others especially for preventing of bio- pollutants with higher efficiency at least for using of animal fecal samples and that contra versa to others (4, 8,12, (Gul , 2007 ) .Using of high Pot. dichromate concentration (5%) do not inhibited the development of Oocysts sporulation or prolonged needed time in this study can be considered as a biological variation or more reliable technique for both of these species of coccidia or the fecal sample of these animal species ( Capra ibex ). The biological pollutants related with this may be not related to other things rather than grown in high rate on hot climate even with using of incubator or other material, and that what was clearly observed within this study.

Conclusion

Local meraz goats infected seriously with many species of coccidia which in some face can be gain special characters .This may be include new species , new variation and new diagnostic modification. All these aspects need more research and attention as these animal mainly stay wild and may affect some epidemiological directions especially in a relation to local goats species which are bred dominantly in this area and for the occurrence of coccidiosis .

Authors’ contributions

Saad MH Al-Bayati is sugested the research idea and write , analysis and managed the results.

Adnan M Al-Rekani is manage samples collection and lab tests in combination with first author .

Ahmed A Hamed is help in samples and lab management.

Table (1): Eimeriosis infection rate of local meriz ( Capra ibex )goats.

Animal age

( month)

Number of animal

Infection rate

Patent period

(days)

+

_

With clinic. Signs

Without clinic. signs

Total

≤ 6 months

37

3

(3.9%)

14

17 (22.08%)

20

55-56±2

≥6 months

40

-

12

12 (15.59%)

28

56-58±2

Total

77

29 ( 37.67%)

48

Table (2): Characteristics features of Eimeria spp. isolated from local meriz ( Capra ibex)goats .

Species number

Measurement*

(Ó®±SE)

Embryonic mass area/L x W ratio

Germinal disk occurrence and measurement

Examined Oocysts

Sporocysts

Number

(Measurement)*

Polar cup

Micro-pyle

Sporullation time(days at room temp.)**

Suspected diagnosis

1

16.03-15.03 (±0.55)

0.42 ±0.09

NC

100

4( 7x8)

-

-

7-10±2

Eimeria alijevi ( fig. 1)

2

24.04 -36.07 (±1.61)

0.18 ±0.08

Clear Outer 1.6x2.08 ± 0.25 Inner 0.83-1.25 ±0.25

100

4( 7x11)

+

+

=

E. christenseni (fig. 2)

3

26.45-31.26 (±2.41)

0.23 ±0.08

=

100

4(8x10)

-

+

=

E.caprovina (fig. 3)

4

28.86 -40.88 (±1.89)

0.18 ±0.09

=

100

NA

+

+

NA

E. minasensis (fig. 4, up)

5

33.67 -40.88 (±0.92)

0.67 ±0.07

=

100

NA

-

+

NA

E.megaembryonica?? ( fig. 4 ,mid)

6

14.34-19.24 (±1.44)

0.39 ±0.09

=

100

NA

-

+

NA

E. ninakohlyakimovae

(fig. 4 , down)

* In Micrometer **Manual agitation NA= Not available L/W=Length/Wide NC=Not clear

Table (3): Sensitivity rate of various diagnostic methods that are used for the diagnosis of Eimeria spp infection in local meriz ( Capra ibex ) goats.

Diagnostic methods

Infection rate

Sensitivity rate

Note

+

-

Direct wet smear preparation

4

25

13.8%

Repeated is essential

Saturated salt solution

27

2

93.1%***

Dry soon and massive flake were seen

Culturing with 2.5 % Pot. Dichromate

10

19

34.5%

Massive fungal growth was observed

Culturing with 5 % Pot. Dichromate

29

zero

100%***

Directly or after floatation

P***=0.001

""

""

Fig. ( 1) : Eimeria alijevi, round Oocyst with no micropyle or cup ( up ) with four elongated sporocysts in full sporulated Oocyst( down ).

""

""

Fig.(2) : Eimeria christenseni , oval Oocyst with micropyle (white arrow )and cup ( down ) with four elongated sporocysts in full sporulated Oocyst and the germinal disk (black arrow ) clear ( up )with outer and inner layer.

""

""

Fig.(3) : Eimeria caprovina oval Oocyst with micropyle (white arrow )and no cup (up) with clear germinal disk ( black arrow ) with outer and inner layer as well as four elongated fat sporocysts ( down ) were observed.

""""

""

Fig. (4) : Eimeria minasensis (up), with cup and micropyle , E.megaembryonia?? (mid) , with no cup but micropyle and E. ninakohlyakimovae ( down )with no cup but micropyle , All species have clear germinal disk with outer and inner layer .