Diversidade taxonômica de <i>Biomphalaria</i> (Planorbidae) no estado de São Paulo, Brasil

Ohlweiler, Fernanda Pires; Rossignoli, Thays de Jesus; Palasio, Raquel Gardini Sanches; Tuan, Roseli

Fernanda Pires Ohlweiler ^**Corresponding author: Fernanda Pires Ohlweiler, e-mail: fpohlweiler@gmail.com

Superintendência de Controle de Endemias, Laboratório de Malacologia da Divisão de Programas Especiais, São Paulo, SP, Brasil.

https://orcid.org/0000-0002-3656-871X

Thays de Jesus Rossignoli

Superintendência de Controle de Endemias, Laboratório de Malacologia da Divisão de Programas Especiais, São Paulo, SP, Brasil.

https://orcid.org/0000-0001-6014-2818

Raquel Gardini Sanches Palasio

Superintendência de Controle de Endemias, Laboratório de Bioquímica e Biologia Molecular, São Paulo, SP, Brasil.

https://orcid.org/0000-0003-1564-0871

Roseli Tuan

Superintendência de Controle de Endemias, Laboratório de Bioquímica e Biologia Molecular, São Paulo, SP, Brasil.

https://orcid.org/0000-0001-7379-783X

*Corresponding author: Fernanda Pires Ohlweiler, e-mail: fpohlweiler@gmail.com

Author Contributions

Fernanda Pires Ohlweiler: Obtaining financial resources from Fapesp. Conceptual and design of the study. Collection, analysis and interpretation of morphological data. Preparation and critical review of the manuscript, including the morphological and molecular study.

Thays de Jesus Rossignoli: Collection and analysis of morphological data. Extraction and amplification of genomic material to obtain COI sequences. Preparation of the manuscript.

Raquel Gardini Sanches Palasio: Performed the DNA Barcoding experiments. Analysis and interpretation of molecular data. Geospatial mapping. Preparation of the manuscript.

Roseli Tuan: Conceptual and practical framework for DNA Barcoding analysis, critical revision, adding intellectual content to molecular analysis.

Conflicts of Interest

The authors declare that they have no conflict of interest related to the publication of this manuscript.

Lot Number	Biomphalaria	CD	Municipality	NM	Latitude	Longitude	Codes for COI sequence	Genbank accession number
615	B. occidentalis	37	Santa Isabel	0	23º17'17" S	46º12'16" W	sti 1	KF926174
		38		0	23º17'00" S	46º12'59" W	sti 5	KF926195
3035		39	Suzano	15	23º33'08" S	46º16'51" W	szn 3,5	MH593417,419
2939	B. oligoza	46	Biritiba Mirim	10	23º36'38" S	46º05'43" W
2989		45		10	23º36'09" S	46º05'13" W
2927		53	Salesópolis	9	23º35'40" S	45º58'41" W
2926	B. peregrina	54	Salesópolis	2	23º32'51" S	45º50'29" W
2991		54		9	23º32'52" S	45º50'26" W	slp 1-5	MH593410-412,428,429
2971	B. schrammi	1	Pirapora do Bom Jesus	10	23º23'22" S	46º59'56" W	pbj 1- 4	MH593405,406, 408,409
2990		1		4	23º23'22" S	46º59'56" W
2645	B. straminea	11	Itapecerica da Serra	4	23º41'09" S	46º48'46" W
2649		11		6	23º41'09" S	46º48'46" W
2661		11		2	23º41'09" S	46º48'46" W
579		38	Santa Isabel	0	23º17'00" S	46º12'59" W	sti 2-4	KF926177, KF926189-190
3000	B. tenagophila	8	Barueri	14	23º30'38" S	46º51'51" W	brr 1-5	MH593423-427
3002		7		16	23º28'49" S	46º52'17" W	brr 6-10	MH593429-433
3015		7		15	23º28'50" S	46º52'17" W
3019		9		14	23º30'53" S	46º49'48" W	brr 11-15	MH593434-438
2931		47	Biritiba Mirim	11	23º33'26" S	46º02'17" W	brt 6,7	MH593398, 399
2932		49		8	23º33'44" S	46º02'35" W	brt 1,2	KF926203,205
2933		48		7	23º33'30" S	46º01'53" W	brt 4,8, 9	MH593396, 400,401
2938		46		2	23º36'39" S	46º05'44" W
2959		51		8	23º33'55" S	46º01'03" W
2996		51		1	23º33'54" S	46º01'03" W
^* * COI sequences, from the Molecular Biology Laboratory of SUCEN, of malacological material not included in the collection of the Divisão de Programas Especiais (DPE) of SUCEN,		52		0	23º33'43" S	46º00'06" W	brt 3	KF926204
2933		50		0	23º33'57" S	46º02'25" W	brt 5	MH593397
1779		12	Embu das Artes	0	23º38'50" S	46º51'11" W	ebt 1	KF926197
		13		0	23º40'08" S	46º51'42" W	ebt 2	KF926198
3054		15	Embu Guaçu	5	23º49'26" S	46º49'24" W	emg 1-4	MH593494,495
								MH593503,504
3055		10	Franco da Rocha	5	23º17'16" S	46º47'54" W	frr 1-7	MH593496-502
3056		10		5	23º17'16" S	46º47'54" W
2936		30	Guarulhos	3	23º24'05" S	46º31'56" W	^ species without COI sequences, CD; Code of the numbers of the collection sites in Figure 1, NM; Number of specimens used in the morphological study.
3021		32		10	23º24'30" S	46º22'40" W	gru 4-6	MH593452-454
3022		33		15	23º24'39" S	46º23'02" W	gru 1-3	MH593449-451
3025		31		15	23º29'36" S	46º32'19" W	gru 7-9	MH593455-557
3034		28		5	23º24'04" S	46º31'56" W	gru 10,11	MH593477, 478
3036		29		5	23º24'17" S	46º31'56" W	gru 12,13	MH593479,480
3023		36	Itaquaquecetuba	13	23º28'48" S	46º21'01" W	itq 1,5	MH593444,448
2918		34	Mairiporã	17	23º19'18" S	46º35'58" W
2968		42	Mogi das Cruzes	15	23º33'25" S	46º15'05" W	mgc 2-4	MH593402,404
2992		43		15	23º34'18" S	46º10'56" W
3001		43		0	23º34'17" S	46º10'56" W	mgc 5-7	MH593420,422
^* * COI sequences, from the Molecular Biology Laboratory of SUCEN, of malacological material not included in the collection of the Divisão de Programas Especiais (DPE) of SUCEN,		44		0	23º34'35" S	46º09'24" W	mgc 1	KF926202
2970		2	Pirapora do Bom Jesus	15	23º23'22" S	46º59'56" W	pbj 3	MH593407
3047		4		4	23º22'35" S	46º59'38" W
3057		3		5	23º24'20" S	47º00'22" W
3053		35	Poá	5	23º33'35" S	46º20'32" W	poa 1,2	MH593505,506
3051		55	Salesópolis	5	23º33'52" S	46º50'08" W	slp 6	MH593493
3052		55		5	23º33'52" S	45º50'02" W
3027		6	Santana de Parnaíba	6	23º25'51" S	46º49'26" W	stp 5-7	MH593496-498
3030		5		7	23º26'17" S	46º56'01" W	stp 1-4	MH593458-461
2148		14	São Lourenço da Serra	0	23º48'11" S	46º55'27" W	sls 1	KF926201
3042		16	São Paulo	5	23º44'35" S	46º43'13" W	^ species without COI sequences, CD; Code of the numbers of the collection sites in Figure 1, NM; Number of specimens used in the morphological study.
3046		17		0	23º46'05" S	46º40'12" W	sao 20-24	MH593488-492
3054		18		0	23º46'36" S	46º39'49" W	sao 13,14	MH593481,482
3037		19		3	23º48'50" S	46º40'02" W
3043		20		5	23º38'20" S	46º37'14" W	sao 15,16	MH593483,484
3044		21		5	23º38'21" S	46º37'09" W	sao 17-19	MH593485-487
3033		22		2	23º31'52" S	46º32'04" W
3041		23		5	23º27'32" S	46º38'31" W	^ species without COI sequences, CD; Code of the numbers of the collection sites in Figure 1, NM; Number of specimens used in the morphological study.
3032		24		10	23º35'33" S	46º26'59" W	sao 5-9	MH593474-478
3031		25		16	23º35'38" S	46º26'59" W	sao 10-12	MH593469-471
3028		26		15	23º36'05" S	46º25'47" W	sao 3,4	MH593464,465
3029		27		10	23º36'09" S	46º25'45" W	sao 1,2	MH593462,463
2997		41	Suzano	3	23º34'15" S	46º17'27" W	szn 1,2	MH593415,416
3001		39		1	23º33'08" S	46º16'51" W	szn 4	MH593418
3016		40		15	23º31'07" S	46º18'18" W	szn 6-10	MH593439-443
Total				452				122

Shell characters	B. tenagophila	B. occidentalis	B. oligoza	B. peregrina	B. schrammi	B. straminea
Color	Brown	Brown	Brown	Brown	Brown	Brown
Diameter (mm)
Average	12,5	7,8	4,2	7	5	6,4
Minimum-Maximum	3,4 - 22,5	6,2 - 13,6	3,3 - 5,2	6,2 - 8,4	4,6 - 5,4	5,2 - 7,0
Width (mm)
Average	5,1	3,5	1,7	2,5	1,7	2,9
Minimum-Maximum	0,7 - 9,6	2,3 - 4,7	1,4 - 2,1	2,0 - 3,0	1,5 - 2,0	2,4 - 3,0
Number of turns	4 1/4 to 7	5 to 6 1/4	4 to 4 1/2	4 1/4 to 4 1/2	4 to 4 3/4	4 1/2 to 5
Aspect of sutures	Defined	Defined	Well defined	Defined	Well defined	Well defined
Shape of the turns^* * morphological characters with expressive anatomical characteristics	Angular	Angular	Rounded	Rounded	Rounded	Rounded
Carenas^* * morphological characters with expressive anatomical characteristics	Present	Present	Absent	Absent	Absent	Absent
Internal lamellas on the last turn of the shell^* * morphological characters with expressive anatomical characteristics	Absent	Absent	Absent	Absent	Present (0 to 6)	Absent
Opening shape^* * morphological characters with expressive anatomical characteristics	Transverse or deltoid	Transverse or deltoid	Rounded	Rounded	Rounded	Rounded
Opening position^* * morphological characters with expressive anatomical characteristics	Front	Front	Front	Lightly deflected to the left	Heavily deflected to the left	Lightly deflected to the left

	B. tenagophila	B. occidentalis	B. oligoza	B. peregrina	B. schrammi	B. straminea
Morphological characters of the mantle
Pattern of mantle pigmentation^* * morphological characters with expressive anatomical characteristics	Tends to be homogeneous	Tends to be homogeneous	Spots or scores	Spots or scores	Spots or scores	Spots or scores
Stain Pattern^* * morphological characters with expressive anatomical characteristics	Inapplicable	Inapplicable	Rounded or misshapen	Rounded or misshapen	elongated, transversely disposed	Rounded or misshapen
Morphological characters of excretory system
Pigmented line in the renal tube	Present in some specimens	Absent	Absent	Absent	Absent	Absent
Oblique salience in the renal tube	Present in some specimens	Absent	Absent	Absent	Absent	Absent
Renal tube	Flat	Flat	Flat	Flat	Flat	Flat
Morphological characters of reproductive system
Number of ovotestis diverticula^* * morphological characters with expressive anatomical characteristics	101 to 429	124 to 176	16 to 38	80 to 113	19 to 30	109 to 143
Aspect of the ovotestis diverticula^* * morphological characters with expressive anatomical characteristics	Little differentiated	Little differentiated	Well differentiated	Well differentiated	Well differentiated	Well differentiated
Shape of the ovotestis diverticula^* * morphological characters with expressive anatomical characteristics	Elongated	Elongated	Bulging	Elongated	Bulging	Elongated
Configuration of the ovotestis diverticula^* * morphological characters with expressive anatomical characteristics	Subdivided and simple	Subdivided and simple	Simple (some subdivided)	Subdivided and simple	Simple (some subdivided)	Subdivided and simple
Aspect of the seminal vesicle^* * morphological characters with expressive anatomical characteristics	Bulky	Reduced	Reduced	Bulky	Reduced	Bulky
Constitution of the conglomerate portion of the seminal vesicle^* * morphological characters with expressive anatomical characteristics	Digitiform expansions	Digitiform expansions	Nodular formations	Digitiform expansions	Digitiform expansions	Digitiform expansions
Aspect of the oviduct pouch^* * morphological characters with expressive anatomical characteristics	Clearly defined	Clearly defined	Bulky	Clearly defined	Discreet	Bulky
Vaginal pouch in the ventral wall of the posterior region of the vagina^* * morphological characters with expressive anatomical characteristics	Present	Absent	Present	Present	Absent	Absent
Aspect of the vaginal pouch^* * morphological characters with expressive anatomical characteristics	Clearly defined	Inapplicable	Discreet	Clearly defined	Inapplicable	Inapplicable
Shape of the vaginal pouch^* * morphological characters with expressive anatomical characteristics	Bulging	Inapplicable	Elongated	Elongated	Inapplicable	Inapplicable
Dilation in the ventral wall of the posterior region of the vagina	Absent	Present in some specimens	Absent	Absent	Absent	Absent
Corrugation on the dorsal wall of the posterior region of the vagina^* * morphological characters with expressive anatomical characteristics	Absent	Absent	Absent	Absent	Absent	Strongly undulated
Extension of the spermatheca in relation to its duct^* * morphological characters with expressive anatomical characteristics	Approximate to the duct	Approximate to the duct	Approximate to the duct	Approximate to the duct	Below the duct	Approximate to the duct
Route of the spermioduto from the genital crossroads to the prostate^* * morphological characters with expressive anatomical characteristics	Winding	Winding	Rectilineal	Rectilineal	Rectilineal	Winding
Shape of the prostate diverticula^* * morphological characters with expressive anatomical characteristics	Tree-like	Tree-like	Simple or subdivided	Tree-like	Tree-like	Tree-like
Number of prostate diverticula^* * morphological characters with expressive anatomical characteristics	10 to 29	18 to 23	0 to 4	14 to 17	12 to 18	12 to 18
Aspect of the penial complex (sheath and prepuce)^* * morphological characters with expressive anatomical characteristics	Robust	Slender	Robust	Robust	Slender	Robust
Width of the penial sheath in relation to the prepuce^* * morphological characters with expressive anatomical characteristics	Narrow (less than 50%)	Narrow (less than 50%)	Wide (greater than 50%)	Wide (greater than 50%)	Wide (greater than 50%)	Wide (greater than 50%)
Extension of the penial sheath in relation to the prepuce^* * morphological characters with expressive anatomical characteristics	Approximate length to the prepuce	Approximate length to the prepuce	Approximate length to the prepuce	Approximate length to the prepuce	Length longer than the prepuce (2 to 4 times)	Length longer than the prepuce
Dimension of the prepuce^* * morphological characters with expressive anatomical characteristics	Wider free termination	Same diameter along its whole length	Same diameter along its whole length	Wider free termination	Subtly wider free termination	Wider free termination

Morphological species	B. tenagophila	B. occidentalis	B. oligoza	B. peregrina	B. schrammi
B. tenagophila	26
B. occidentalis	6	26
B. oligoza	0	1	26
B. peregrina	1	0	1	26
B. schrammi	0	1	3	0	26
B. straminea	0	0	1	1	1

Morphological species	N	H	Hd	π	K
B. tenagophila	106	6	0.52±0.05	0.01	5.564
B. straminea	3	2	0.66±0.31	0.01	8
B. occidentalis	4	1	0	0	0
B. peregrina	5	1	0	0	0
B. schrammi	4	1	0	0	0
Biomphalaria spp.	122	11	0.64±0.04	0.03	16.34

Morphological species	B.tenagophila	B.occidentalis	B.straminea	B.schrammi	B.peregrina
B.tenagophila	0-0.03
B.occidentalis	0.05	0-0.00
B.straminea	0.09	0.09	0-0.01
B.schrammi	0.12	0.11	0.12	0-0.00
B.peregrina	0.12	0.11	0.10	0.13	0-0.00

Morphological species	NS	GenBank sequences	GenBank best-fit	GenBank similarity	NH	ABGD	bPTP	GMYC
B. tenagophila	106	106	B. tenagophila	99-100%	6	1	1	5
B. straminea	3	96	B. straminea	99-100%	2	1	1	1
B. occidentalis	4	21	B. occidentalis	99-100%	1	1	1	1
B. peregrina	5	14	B. peregrina	97-98%	1	1	1	1
B. schrammi	4	0	Planorbidae	91%	1	1	1	1
NPS						5	5	9

[1] *Corresponding author: Fernanda Pires Ohlweiler, e-mail: fpohlweiler@gmail.com

Brasil

Brasil

Diversidade taxonômica de Biomphalaria (Planorbidae) no estado de São Paulo, Brasil

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