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Morphological and molecular characterization of Butlerius butleri Goodey, 1929 (Nematoda: Diplogastridae) from South Africa: First report

Chantelle Girgan

Chantelle Girgan

Nematology Unit, Biosystematics, ARC-Plant Health and ProtectionQueenswood, South Africa
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Girgan, Chantelle
, 
Gerhard Du Preez

Gerhard Du Preez

Unit for Environmental Sciences and Management, North-West UniversityPotchefstroom, South Africa
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Du Preez, Gerhard
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Hendrika Fourie

Hendrika Fourie

Unit for Environmental Sciences and Management, North-West UniversityPotchefstroom, South Africa
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Fourie, Hendrika
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Milad Rashidifard

Milad Rashidifard

Unit for Environmental Sciences and Management, North-West UniversityPotchefstroom, South Africa
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Rashidifard, Milad
  
01 gen 2021
Morphological and molecular characterization of Butlerius butleri Goodey, 1929 (Nematoda: Diplogastridae) from South Africa: First report's Cover Image
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Pubblicato online: 01 gen 2021
Pagine: 1 - 12
DOI: https://doi.org/10.21307/jofnem-2021-026
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© 2021 Authors, published by Sciendo.
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Figure 1:

Butlerius butleri. A: Female anterior region; B: Female stoma; C: Male stoma; D: Vulval region indicating four large glands opening into proximal part of uterus; E: Anterior branch of female genital tract; F: Male posterior region indicating genital papillae; G: Female tail; H: Female habitus; I: Male habitus. Scale bars: B to D, F, H: 10 µm; A, E: 20 µm; G: 50 µm; H, I: 100 µm.
Butlerius butleri. A: Female anterior region; B: Female stoma; C: Male stoma; D: Vulval region indicating four large glands opening into proximal part of uterus; E: Anterior branch of female genital tract; F: Male posterior region indicating genital papillae; G: Female tail; H: Female habitus; I: Male habitus. Scale bars: B to D, F, H: 10 µm; A, E: 20 µm; G: 50 µm; H, I: 100 µm.

Figure 2:

Light microscope pictures of females of Butlerius butleri Goodey, 1929 from South Africa. A: Anterior region with stoma, arrow indicating excretory pore; B: Stoma; C: Base of corpus showing swollen metacorpus and junction with isthmus, arrow indicating excretory pore; D: Vulval region and posterior genital tract; E: Vulval region showing schlerotised dorsal wall of uterus and four large glands; F: Tail region cuticle with punctations and position of phasmid; G: Tail. Scale bars: B, C, E, F: 10 µm; A, D, G: 20 µm.
Light microscope pictures of females of Butlerius butleri Goodey, 1929 from South Africa. A: Anterior region with stoma, arrow indicating excretory pore; B: Stoma; C: Base of corpus showing swollen metacorpus and junction with isthmus, arrow indicating excretory pore; D: Vulval region and posterior genital tract; E: Vulval region showing schlerotised dorsal wall of uterus and four large glands; F: Tail region cuticle with punctations and position of phasmid; G: Tail. Scale bars: B, C, E, F: 10 µm; A, D, G: 20 µm.

Figure 3:

Light microscope pictures of males of Butlerius butleri Goodey, 1929 from South Africa. A: Stoma; B: Excretory pore; C: Anterior region showing cuticle with punctations and position of amphid; D: Cloacal region indicating cloacal flap, v5,6,7 clusters and posterior dorsal papilla; E: Cloacal region showing spicule and gubernaculum; F: Sphincter in mid-region of vas deferens. Scale bars: A to F: 10 µm.
Light microscope pictures of males of Butlerius butleri Goodey, 1929 from South Africa. A: Stoma; B: Excretory pore; C: Anterior region showing cuticle with punctations and position of amphid; D: Cloacal region indicating cloacal flap, v5,6,7 clusters and posterior dorsal papilla; E: Cloacal region showing spicule and gubernaculum; F: Sphincter in mid-region of vas deferens. Scale bars: A to F: 10 µm.

Figure 4:

Bayesian phylogenetic tree with 50% majority rule of Butlerius butleri from South Africa using small subunit (SSU) rDNA gene sequences under GTR + G model (lnL = 7,712.8957; K = 117; freqA = 0.2568; freqC = 0.1991; freqG = 0.2542; freqT = 0.2900; rAC = 0.9137; rAG = 2.6380; rAT = 1.9015; rCG = 0.2936; rCT = 4.5476; rGT = 1.0000; gamma shape = 0.3330). The newly obtained sequences are indicated by bold font.
Bayesian phylogenetic tree with 50% majority rule of Butlerius butleri from South Africa using small subunit (SSU) rDNA gene sequences under GTR + G model (lnL = 7,712.8957; K = 117; freqA = 0.2568; freqC = 0.1991; freqG = 0.2542; freqT = 0.2900; rAC = 0.9137; rAG = 2.6380; rAT = 1.9015; rCG = 0.2936; rCT = 4.5476; rGT = 1.0000; gamma shape = 0.3330). The newly obtained sequences are indicated by bold font.

Figure 5:

Bayesian phylogenetic tree with 50% majority rule of Butlerius butleri from South Africa using Large subunit (LSU) rDNA gene sequences under GTR + I + G model (lnL = 11,826.2635; K = 111; freqA = 0.1404; freqC = 0.2230; freqG = 0.3544; freqT = 0.2822; rAC = 0.8351; rAG = 2.7011; rAT = 1.4534; rCG = 0.5981; rCT = 4.6690; rGT = 1.0000; gamma shape = 0.5020). The newly obtained sequences are indicated by bold font.
Bayesian phylogenetic tree with 50% majority rule of Butlerius butleri from South Africa using Large subunit (LSU) rDNA gene sequences under GTR + I + G model (lnL = 11,826.2635; K = 111; freqA = 0.1404; freqC = 0.2230; freqG = 0.3544; freqT = 0.2822; rAC = 0.8351; rAG = 2.7011; rAT = 1.4534; rCG = 0.5981; rCT = 4.6690; rGT = 1.0000; gamma shape = 0.5020). The newly obtained sequences are indicated by bold font.

Polymerase chain reaction steps used for amplification of the SSU and LSU rDNA genes_

35 cycles
Primers Initial denaturation Denaturation Annealing Extension Final extension
SSU F04/SSU R26 94°C 3 min 94°C 45 sec 54°C 45 sec 72°C 45 sec 72°C 6 min
D2A/D3B 94°C 3 min 94°C 45 sec 56°C 45 sec 72°C 45 sec 72°C 6 min

Morphometrics of two populations of Butlerius butleri Goodey, 1929 females and males found in compost heaps in Potchefstroom, South Africa_

Population 1 Population 2
Characteristics Female (n = 7) Male (n = 10) Female (n = 3) Male (n = 2)
L 1,315 ± 90.2 (1,166-1,397) 1,193 ± 110.1 (1,082-1,365) 1,423.0* –, 1,399
L′ 979 ± 75.2 (880-1,086) 887 ± 68.2 (783-1,009) 1,140 ± 69.7 (1,093-1,220) 1,148, 1,136
a 44.5 ± 2.7 (40.8-47.6) 47.6 ± 6.0 (37.5-54.2) 39.9* –, 43.4
a′ 32.5 ± 2.2 (30.2-35.3) 35.3 ± 3.3 (29.3-39.6) 32.8 ± 1.9 (31.0-34.8) 36.8, 35.3
b 5.3 ± 0.4 (4.7-5.8) 5.2 ± 0.2 (4.9-5.4) 5.5* –, 5.4
b′ 3.9 ± 0.3 (3.5-4.1) 3.9 ± 0.2 (3.5-4.3) 4.3 ± 0.2 (4.1-4.5) 4.8, 4.4
c 4.6 ± 0.9 (4.0-6.0) 3.9 ± 0.4 (3.4-4.6) 4.5* –, 5.3
c′ 14.5 ± 2.0 (11.7-16.3) 13.5 ± 3.1 (8.7-16.9) 13.3* –, 9.2
V/T 45.7 ± 1.3 (43.5-46.6) 39.2 ± 3.2 (36.3-44.4) 45.3* –, 50.2
G1 12.6 ± 1.4 (11.4-14.8) – 13.0* –, –
G2 12.4 ± 0.9 (11.5-13.5) – 13.8* –, –
Body width at midbody 31 ± 3.1 (26-35) 25 ± 2.9 (20-30) 35 ± 1.1 (34-36) 31, 32
Labial region diameter 19 ± 1.9 (18-23) 17 ± 1.7 (14-20) 19 ± 0.7 (18-19) 18, 19
Cephalic setae length 8 ± 0.7 (8-9) 8 ± 0.9 (7-10) 7 ± 0.5 (7-8) 7, 7
Length of stoma 26 ± 1.5 (23-28) 24 ± 1.7 (21-27) 27 ± 0.6 (27-28) 26, 26
Stoma width 11 ± 1.1 (9-13) 10 ± 1.3 (8-12) 12 ± 0.9 (12-13) 12, 12
Cheilostom length 12 ± 0.8 (11-13) 10 ± 2.2 (5-13) 14 ± 1.6 (12-15) 13, 12
Gymnostom length 7 ± 1.0 (6-9) 7 ± 2.0 (5-10) 6 ± 0.4 (6-7) 6, 6
Stegostom length 7 ± 1.1 (6-9) 5 ± 1.1 (4-7) 7 ± 0.8 (6-8) 7, 8
Dorsal tooth length 9 ± 1.2 (7-10) 7 ± 0.9 (6-8) 7 ± 0.7 (6-7) 9, 7
Subventral tooth length 5 ± 0.9 (4-6) 4 ± 0.4 (4-5) 5 ± 0.7 (4-5) 5, 4
Corpus (procorpus and metacorpus) 137 ± 3.2 (133-141) 125 ± 7.2 (113-140) 144 ± 2.5 (142-147) 138, 134
Postcorpus (isthmus and basal bulb) 115 ± 5.3 (107-120) 102 ± 8.6 (86-118) 121 ± 5.2 (116-126) 102, 125
Pharynx (anterior end to base of basal bulb) 252 ± 8.1 (241-262) 227 ± 15.3 (199-258) 265 ± 6.4 (258-269) 240, 259
Excretory pore from anterior 191 ± 45.4 (163-243) 156 ± 9.4 (146-171) 157 ± 2.5 (155-158) –, 173
Nerve ring from anterior 140 ± 6.3 (133-149) 123 ± 10.6 (100-140) 146 ± 3.5 (143-150) 138, 134
Metacorpus width 22 ± 2.5 (19-26) 20 ± 1.2 (18-22) 28 ± 3.7 (25-32) 20, 23
Basal bulb width 22 ± 2.6 (18-25) 18 ± 1.5 (16-21) 27 ± 0.6 (27-28) 18, 22
Cardia length 9 ± 1.3 (8-12) 9 ± 3.1 (6-14) – 7, –
Anterior genital tract length 166 ± 27.8 (129-205) – 196 ± 11.8 (185-209) –
Posterior genital tract length 157 ± 15.2 (133-184) – 188 ± 7.1 (182-196) –
Body width at vulva 31 ± 3.2 (26-36) – 35 ± 1.5 (34-36) –
Vulva from anterior end 608 ± 42.5 (542-671) – 683 ± 49.5 (645-739) –
Vulva-anus distance 376 ± 32.1 (338-415) – 457 ± 27.2 (427-481) –
Vagina length 12 ± 2.5 (10-16) – 14 ± 2.1 (12-16) –
Rectum length 32 ± 2.6 (29-36) – 31 ± 5.3 (28-37) –
Body width at anus/cloaca 20 ± 1.6 (18-22) 24 ± 3.2 (19-28) 24 ± 0.4 (24-25) 26, 29
Testis length – 496 ± 72.0 (392-589) – 607, 703
Spicules length – 40 ± 2.7 (36-43) – 39, 43
Gubernaculum length – 27 ± 2.6 (23-31) – 30, 29
Tail length 290 ± 46.4 (225-325) 310 ± 43.2 (242-356) 317.0 –, 263
Phasmid posterior to anus 29 ± 1.5 (28-31) 36 ± 4.3 (30-44) – –, 37
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