Prevalence and Molecular Characterization of Ascaridoid Parasites of Philippine Decapterus species

Tres Tinna M. Dela Cruz; Kennesa Klariz R. Llanes; Joliesa Mae S. Toledo; Junard A. Catabay; Raffy Jay C. Fornillos; Ian Kendrich C. Fontanilla; Vachel Gay V. Paller

doi:10.2478/jofnem-2022-0030

Figures & Tables

Fish landing area at Balayan Bay and Tayabas Bays, southern Luzon, Philippines, for fish sample collection.

Light microscopic images of representative marine ascaridoid larvae collected from Decapterus spp. A and D, anterior part of the body, ventral view, showing the larval tooth (T), esophagus (E), and ventriculus (V); B and E (inset), detailed larval tooth (T); C and F, conical tail showing or lacking a mucron (M), respectively.

Prevalence (%) and mean intensity (worms/host ± SD) of marine ascaridoid larvae across the three Decapterus spp. (N = 371) collected from Tayabas Bay and Balayan Bay, southern Luzon, Philippines.

Aggregated distribution of marine ascaridoid larval parasites from (A) pooled Decapterus spp. samples, (B) Decapterus maruadsi, (C) Decapterus tabl, and (D) Decapterus macrosoma from Tayabas Bay and Balayan Bay, southern Luzon, Philippines.

Intra- and interspecific sequence variations in Anisakidae and Raphidascarididae obtained in the GenBank and sample isolate sequences. (A) 18S rRNA and (B) 5.8S-ITS2-28S regions.

ML phylogenetic tree of Anisakidae and Raphidascarididae inferred from 628 nucleotide positions of the 18S rRNA gene using the TrNef + G model of DNA substitution and rooted on Ascaris lumbricoides (U94366). Values on nodes represent ML/NJ bootstrap percentages of 1,000 bootstrap samples; values <50% are not shown. Scale bar represents five nucleotide substitutions of 1,000 nucleotides. Sequences from this study are highlighted. ML, maximum-likelihood; NJ, neighbor-joining.

ML phylogenetic tree of Anisakidae and Raphidascarididae inferred from 460 nucleotide positions from the partial sequence of the 5.8S, whole sequence of the ITS2, and partial sequence of 28S rRNA gene using the HKY + G model of DNA substitution and rooted on Raphidascaris (Sprentascaris) andersoni. Values on nodes represent ML/NJ bootstrap percentages of 1,000 bootstrap samples; values <50% are not shown. Scale bar represents two nucleotide substitutions of 10 nucleotides. Sequences from this study are highlighted. ITS2, internal transcribed spacer 2; ML, maximum-likelihood.

GenBank sequences of close species matches used for phylogeny construction_

Ascaridoid species	Host	Habitat	Geographical Origin	ITS2	18S rRNA
Anisakis pegreffii	Trachinus radiatus	Marine	Tunisia	MT820022	-
Anisakis pegreffii	Conger myriaster	Marine	China	-	MF072697
Anisakis pegreffii	Caretta	Marine	Italy	-	EF180082
Anisakis simplex	Globicephala melaena	Marine	Norway	AY826723	-
Anisakis simplex	Scomber japonicus	Marine	Japan	LC621351	-
Anisakis simplex	Carcharhinus sorrah	Marine	China	-	MF072711
Anisakis typica	Tursiops aduncus	Marine	Egypt	HF911524	-
Anisakis typica	Bottlenose dolphin	Marine	China	KF673776	-
Anisakis typica	Rastrelliger kanagurta	Marine	Thailand	AB432909	-
Anisakis typica	Stenella longirostris	Marine	Brazil	AY826724	-
Anisakis typica	Katsuwonus pelamis	Marine	Indonesia	KC928262	-
Anisakis typica	Sotalia fluviatilis	Marine	Brazil	EU327686	-
Anisakis ziphidarum	Pagellus bogaraveo	Marine	Portugal	JN005767	-
Anisakis ziphidarum	Hoplostethus cadenati	Marine	Mauritius	EU718473	-
Anisakis ziphidarum	Ziphius cavirostris	Marine	South Africa	AY826725	-
Contracaecum eudyptulae	Eudyptula minor	Marine	Australia	-	EF180072
Contracaecum microcephalum	NM	NM	NM	-	AY702702
Contracaecum multipapillatum	NM	NM	NM	-	U94370
Hysterothylacium aduncum	Zoarces viviparus	Marine	Denmark	JX845137	-
Hysterothylacium aduncum	Dolphin fish	Marine	South Korea	HQ702733	-
Hysterothylacium aduncum	Lophius litulon	Marine	China	-	MF072693
Hysterothylacium deardorffoverstreetorum	Paralichthys isosceles	Marine	Brazil	-	JF718550
Hysterothylacium fortalezae	NM	NM	NM	-	U94374
Hystetothylacium reliquens	NM	NM	NM	-	U94376
Hystetothylacium tetrapteri	Kajikia audax	Marine	China	-	MF072705
Hysterothylacium thalassini	Priacanthus macracanthus	Marine	China	-	MF072702
Pseudanisakis rajae	Raja pulchra	Marine	China	-	MF072707
Pseudoterranova decipiens	Gadus morhua	Marine	Denmark	KM273088	-
Pseudoterranova decipiens	NM	NM	NM	-	U94380
Raphidascaroides brasiliensis	Platydoras costatus	Freshwater	Brazil	-	KP726276
Raphidascaroides moraveci	Platydoras armatulus	Freshwater	Brazil	-	KP726278
Raphidascaris acus	Esox lucius	Freshwater; brackish water	Finland	-	DQ503460
Raphidascaris lanfredae	Geophagus maximus	Freshwater	Brazil	-	KX859077
Raphidascaris longispicula	NM	Marine	South China Sea	JN102362	-
Raphidascaris longispicula	Uroconger lepturus	Marine	China	KP326546	MF072704
Raphidascaris lophii	NM	NM	China	MF422212	-
Raphidascaris lophii	Lophius litulon	Marine	China	MH211584	MF072692
Raphidascaris lophii	NM	Marine	China	JF809816	-
Terranova caballeroi	NM	NM	NM	-	U94381
^aRaphidascaris (Sprentascaris) andersoni	Gymnogeophagus balzanii	Freshwater	Brazil	MK141032	-
^aAscaris lumbricoides	NM	NM	NM	-	U94366

BLAST results for the 18S rRNA and 5_8S-ITS2-28S sequences of ascaridoid samples_

Specimen	18S rRNA		5.8S-ITS2-28S		Family
accession (18S/ITS2)	GenBank accession	% Match	GenBank accession	% Match	Family
OK659774/ OK659786	MF072704 (Raphidascaris longispicula)	99.84	MF422212 (Raphidascaris longispicula)	99.18	Raphidascarididae
OK659775/ OK659787	MF072711 (Anisakis simplex) MF072697 (Anisakis pegreffii) U94365 (Anisakis sp.)	99.52	HF911524 (Anisakis simplex)	99.88	Anisakidae
OK659776/ OK659788	MF072704 (Raphidascaris longispicula)	99.84	MF422212 (Raphidascaris longispicula)	99.79	Raphidascarididae
OK659777/ OK659789	MF072711 (Anisakis simplex) MF072697 (Anisakis pegreffii) U94365 (Anisakis sp.)	99.68	HF911524 (Anisakis simplex)	99.88	Anisakidae
OK659778/ OK659790	MF072704 (Raphidascaris longispicula)	99.52	MF422212 (Raphidascaris longispicula)	99.79	Raphidascarididae
OK659779/ OK659791	MF072704 (Raphidascaris longispicula)	99.52	MF422212 (Raphidascaris longispicula)	99.59	Raphidascarididae
OK659780/ OK659792	MF072711 (Anisakis simplex) MF072697 (Anisakis pegreffii) U94365 (Anisakis sp.)	99.20	HF911524 (Anisakis simplex)	98.66	Anisakidae
OK659781/ OK659793	MF072711 (Anisakis simplex) MF072697 (Anisakis pegreffii) U94365 (Anisakis sp.)	99.68	HF911524 (Anisakis simplex)	98.79	Anisakidae
OK659782/ OK659794	MF072711(Anisakis simplex) MF072697 (Anisakis pegreffii) U94365 (Anisakis sp.)	99.52	HF911524 (Anisakis simplex)	99.87	Anisakidae
OK659783/ OK659795	MF072711(Anisakis simplex) MF072697 (Anisakis pegreffii) U94365 (Anisakis sp.)	99.68	HF911524 (Anisakis simplex)	98.86	Anisakidae
OK659784/ OK659796	MF072711 (Anisakis simplex) MF072697 (Anisakis pegreffii) U94365 (Anisakis sp.)	99.68	HF911524 (Anisakis simplex)	98.61	Anisakidae
OK659785/ OK659797	MF072711 (Anisakis simplex) MF072697 (Anisakis pegreffii) U94365 (Anisakis sp.)	99.68	HF911524 (Anisakis simplex)	99.87	Anisakidae

Corrected (TrNef + G) genetic distances of 18S rRNA gene and corrected (HKY + G) genetic distances of the 5_8S-ITS2-28S region of Anisakidae and Raphidascarididae nematodes_

Ascaridoid comparisons	Number of pairwise comparisons	18S rRNA		5.8S-ITS2-28S region
Ascaridoid comparisons	Number of pairwise comparisons	Minimum	Maximum	Minimum	Maximum
Intraspecific	67 (18S), 118 (ITS2)	0	0.0182098	0	0.0077422
Interspecific	494 (18S), 410 (ITS2)	0	0.04259699	0.00493606	1.8776778

Prevalence and Molecular Characterization of Ascaridoid Parasites of Philippine Decapterus species

Figures & Tables

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GenBank sequences of close species matches used for phylogeny construction_

BLAST results for the 18S rRNA and 5_8S-ITS2-28S sequences of ascaridoid samples_

Corrected (TrNef + G) genetic distances of 18S rRNA gene and corrected (HKY + G) genetic distances of the 5_8S-ITS2-28S region of Anisakidae and Raphidascarididae nematodes_

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