Ultramorphology and molecular studies of Contracaecum larvae (Nematoda: Anisakidae) collected in five Cyprinid fish species from Sulaimani Province, Kurdistan Region-Iraq

Abdullah, Y. S.; Abdullah, S. M. A.; Hussein, R. H.

doi:10.2478/helm-2021-0001

Figures & Tables

Map of north Iraq showing the study area.

Photo micrograph of Contracaecum larva in Carasobarbus luteus. A - Anterior region of the larva (I00×); B - Posterior region of the larva (I00×); C - Mouth region of the larva (400×); D - Tail region of the larva (400×) an = anus; bt = boring tooth; er = cuticle ridges; eso = esophagus; esoc = esophageal caecum; int = intestine; intc = intestinal caecum; mo = mouth opening; p = papillae; rec = rectum; ta = tail.

Scanning electron micrograph of Contracaecum larva in Carasobarbus luteus. A - Anterior region of the larva; B - Head region of the larva; C - Tail region of the larva; D - Cuticle of the larva an = anus; bt = boring tooth; cu = cuticle; exp = excretory pore; mo = mouth opening; p = papillae; ta = tail.

Multiple sequence alignment for ITS-1 in C. rudolphii B in the five different fish species.

Multiple sequence alignment for ITS-2 in C. rudolphii B in the five different fish species.

Alignment of the ITS-1 and ITS-2 sequences representing genotype 1 (C. rudolphii B) from the present study and genotype 2 (C. rudolphii A) sequences have been deposited in GenBank under the accession numbers AJ634 782 and AJ634 785. Nucleotide differences between the aligned equences are indecated by having no asterisks.

Phylogenetic relationships between Contracaecum larvae from the present study and other Contracaecum species as inferred by maximum likelihood obtained from ITSl. Ascaris suum was used as outgroup.

Contracaecum nematodes and specimens/accession number (ITS) of taxa used to build phylogenetic trees and the nucleotide (bp) differences with the Contracaecum larvae collected from the present study_

Parasite	GeneBank accession no. (ITS)	No. of nucleotide differences (bp)	Host	Source
C. rudolphii A	ITS1 (AJ634782) ITS2 (AJ634785)	9 14	Phalacrocorax carbo sinensis Phalacrocorax carbo sinensis	Li et al. (2005) Li et al. (2005)
C. rudolphii B	ITS1 (AJ634783) ITS2 (AJ634786)	1 0	Phalacrocorax carbo sinensis Phalacrocorax carbosinensis	Zhang et al., 2009 Li et al. (2005)
C. rudolphii D	ITS1 (FM210251) ITS2 (FM210268)	6 17	Phalacrocorax varius Phalacrocorax carbo sinensis	Shamsi et al. (2009b) Shamsi et al. (2009b)
C. rudolphii E	ITS1 (FM210257) ITS2 (FM210271)	6 15	Phalacrocorax varius Phalacrocorax varius	Shamsi et al. (2009b) Shamsi et al. (2009b)
C. rudolphii F	ITS (JF424597)	21 16	Pelecanus occidentalis Pelecanus occidentalis	D'Amelio et al. (2012) D'Amelio et al. (2012)
C. ogmorhini	ITS1 (AJ291468) ITS2 (AJ291471)	10 15	Arctocephalus pusillus doriferus Zalophus californianus	Zhu et al. (2001) Zhu et al. (2001)
C. eudyptulae	ITS1 (AJ007461) ITS2 (FM177565)	8 17	- Eudyptula minor	Zhu et al.(unpublished) Shamsi et al. (2009a)
C. chubutensis	ITS1 (HQ389546) ITS2 (HQ389548)	13 23	Phalacrocorax brasilianus Phalacrocorax atriceps	Garbin et al. (2011) Garbin et al. (2011)
C. variegatum	ITS1 (MK424804)	22	Bird	Hbaiel & Mohammad (unpublished)
C. microcephalum	ITS2 (FM177537) ITS1 (FM177523) ITS2 (FM177527)	14 50 72	Anhinga melanogaster Phalacrocorax melanoleucos Phalacrocorax melanoleucos	Shamsi et al. (2009a) Shamsi et al. (2009a) Shamsi et al. (2009a)
C. septentrionale	ITS1 (AJ634784) ITS2 (AJ634787)	24 35	Phalacrocorax carbo sinensis Alca torda	Li et al. (2005) Li et al. (2005)
C. bioccai	ITS (JF424598)	30 40	Pelecanus occidentalis Pelecanus occidentalis	D'Amelio et al. (2012) D'Amelio et al. (2012)
C. radiatum	ITS (AY603529)	35 65	Leptonechotes weddlli Leptonechotes weddlli	Kijewska et al. (2008) Kijewska et al. (2008)
C. osculatum	ITS (AB277825)	77 105	Arabesque greenling Arabesque greenling	Umehara et al. (2008) Umehara et al. (2008)
C. multipapillatum	ITS1 (AM940056) ITS2 (AM940060)	146 132	Pelecanus conspicillatus Pelecanus conspicillatus	Shamsi et al. (2008) Shamsi et al. (2008)
C. pyripapillatum	ITS1 (AM940062) ITS2 (AM940066)	141 136	Pelecanus conspicillatus Pelecanus conspicillatus	Shamsi et al. (2008) Shamsi et al. (2008)
Contracaecum larva	ITS1 (MN557376) ITS2 (MN526259)	- -	Acanthobrama marmid Acanthobrama marmid	Present study Present study
Contracaecum larva	ITS1 (MN557377) ITS2 (MN563727)	- -	Arabibarbus grypus Arabibarbus grypus	Present study Present study
Contracaecum larva	ITS1 (MN557378) ITS2 (MN563728)	- -	Capoeta trutta Capoeta trutta	Present study Present study
Contracaecum larva	ITS1 (MN557379) ITS2 (MN563729)	- -	Carasobarbus luteus Carasobarbus luteus	Present study Present study
Contracaecum larva	ITS1 (MN557380) ITS2 (MN563730)	- -	Chondrostoma regium Chondrostoma regium	Present study Present study
Ascaris suum	ITS1 (AB110023) ITS2 (FJ418786)	- -	Pig Pig	Ishiwata et al. (2004) Wickramasinghe et al. (2009)

Prevalence of Contracaecum larva and mean of intensity among infected fish species_

Host	Fish		Prevalence	Mean intensity
	Examined	Infected	%
Acanthobrama marmid	20	7	35.00	5.57
Arabibarbus grypus	123	1	0.81	7.00
Capoeta trutta	222	2	0.90	3.00
Carasobarbus luteus	89	4	4.49	4.25
Chondrostoma regium	52	3	5.76	9.00

Contracaecum nematodes and specimens/accession number (COX-2) of taxa used to build phylogenetic trees and the nucleotide (bp) differences with the Contracaecum larvae collected from the present study_

Parasite	GeneBank accession no. (COX-2)	No. of nucleotide differences (bp)	Host	Source
C. rudolphii A	EF122201	34	Phalacrocorax carbo sinensis	Mattiucci et al. (2008)
C. rudolphii B	EF558894	1	Phalacrocorax carbo sinensis	Mattiucci et al. (2008)
C. rudolphii C	EF014283	272	Phalacrocorax auritus	D'Amelio et al. (2007)
C. rudolphii F	JF727879	40	Pelecanus occidentalis	D'Amelio et al. (2012)
C. ogmorhini	MN624184	32	Zalophus californianus	Mladineo et al. (under press)
C. chubutensis	HQ328504	46	Phalacrocorax atriceps	Garbin et al. (2011)
C. microcephalum	EF122208	71	Phalacrocorax pygmaeus	Mattiucci et al. (2008)
C. septentrionale	EF558898	60	Phalacrocorax carbo carbo	Mattiucci et al. (2008)
C. bioccai	EF558899	50	Pelecanus occidentalis	Mattiucci et al. (2008)
C. osculatum	KC412224	58	Chionodraco hamatus	Santoro et al. (2013)
C. multipapillatum	AF179910	72	-	Nadler & Hudspeth (2000)
C. micropapillatum	EU852350	70	Pelecanus onocrotalus	Mattiucci et al. (2010)
C. austral	GQ847539	55	Phalacrocorax brasilianus	Garbin et al. (2011)
C. pelagicum	EF122210	60	Spheniscus magellanicus	Mattiucci et al. (2008)
Contracaecum larva	MN589997	-	Acanthobrama marmid	Present study
Contracaecum larva	MN589998	-	Arabibarbus grypus	Present study
Contracaecum larva	MN589999	-	Capoeta trutta	Present study
Contracaecum larva	MN590000	-	Carasobarbus luteus	Present study
Contracaecum larva	MN590001	-	Chondrostoma regium	Present study
Ascaris suum	HQ704901	-	swine	Liu et al. (2012)

Comparison of measurements in Contracaecum larvae in different infected fish species in the present study (in millimeter)_

Host	Total length	Maximum width	Tail length	Rectum length	Boring teeth length	Esophagus length	Esophageal caeca length	Intestinal caeca length
A. marmid	4.10	0.20	0.07	0.07	0.005	0.480	0.50	0.38
A. grypus	5.07	0.25	0.07	0.07	0.010	0.650	0.30	0.20
C. trutta	4.20	0.23	0.08	0.07	0.005	0.450	0.48	0.33
C. luteus	4.62	0.25	0.08	0.07	0.005	0.600	0.55	0.40
C. regium	7.75	0.35	0.08	0.09	0.010	0.600	0.60	0.45

Accession numbers provided by NCBI for the collected Contracaecum larvae in different fish hosts in the present study_

Host of Contracaecum rudolphii	Accession numbers for ITS-1 sequences	Accession numbers for ITS-2 sequences	Accession numbers for COX-2
Acanthobrama marmid	MN557376	MN526259	MN589997
Arabibarbus grypus	MN557377	MN563727	MN589998
Capoeta trutta	MN557378	MN563728	MN589999
Carasobarbus luteus	MN557379	MN563729	MN590000
Chondrostoma regium	MN557380	MN563730	MN590001

Ultramorphology and molecular studies of Contracaecum larvae (Nematoda: Anisakidae) collected in five Cyprinid fish species from Sulaimani Province, Kurdistan Region-Iraq

Figures & Tables

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Contracaecum nematodes and specimens/accession number (ITS) of taxa used to build phylogenetic trees and the nucleotide (bp) differences with the Contracaecum larvae collected from the present study_

Prevalence of Contracaecum larva and mean of intensity among infected fish species_

Contracaecum nematodes and specimens/accession number (COX-2) of taxa used to build phylogenetic trees and the nucleotide (bp) differences with the Contracaecum larvae collected from the present study_

Comparison of measurements in Contracaecum larvae in different infected fish species in the present study (in millimeter)_

Accession numbers provided by NCBI for the collected Contracaecum larvae in different fish hosts in the present study_

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