With respect to the isolation of soil nematodes using the
During a survey of soil nematodes in Meerut, Uttar Pradesh, India, two isolates of
Soil samples were collected from agricultural farmlands in Mawana, Meerut (28°9´N, 77°71´E, and elevation of 225 m), India, and were tested for the presence of nematodes. Nematode specimens were isolated from two soil samples by
Nematode specimens were observed for morphological characters under phase contrast microscope (Nikon Eclipse 50i) and light microscope (Magnus MLX) while morphometric characters were measured with built-in software of the Nikon Eclipse 50i (Nikon DS–L1). Demanian indices (de Man, 1880) and other morphometrical ratios were calculated. Line drawings were made with the help of drawing tube attached to the Nikon microscope provided with differential interference contrast (DIC) optics. Images were taken with the Nikon microscope that was provided with DIC optics and Nikon Digital Sight DS-U1 camera. Micrographs were edited using Adobe® Photoshop® CS. The terminology used for the morphology of stoma and spicules follows the proposals by De Ley et al. (1995) and Abolafia and Peña-Santiago (2017a), respectively.
For the SEM, male and female generations were first fixed in TAF and then preserved in glycerine. Glycerine preserved specimens were used for SEM observations according to the Abolafia’s (2015) proctocol. They were hydrated in distilled water, dehydrated in a graded ethanol-acetone series, critical point dried with liquid CO2, mounted on SEM stubs and finally coated with gold. The mounts were examined with a Zeiss Merlin microscope (5 kV) (Zeiss, Oberkochen, Germany).
DNA was extracted from pool of juveniles isolated from cadavers of
The sequences were edited and compared with those already present in GenBank using the basic local alignment search tool (BLAST) of the National Centre for Biotechnology Information (NCBI) (Altschul et al., 1990). An alignment of nematode samples together with sequences of related cephalobid species was produced for the LSU (D2-D3 rDNA), SSU, and ITS rDNA sequences using default Clustal W parameters in MEGA 6.0 (Kumar et al., 2016) and optimized manually in BioEdit (Hall, 1999). Pairwise distances were computed using MEGA 6.0 (Kumar et al., 2016). All characters were treated as equally weighted and gaps as missing data.
The morphological and morphometrical studies and molecular (D2-D3, 18 S and ITS rDNA) analyses confirmed the present strains KMW and DH1 as conspecific to
Material examined: 20 females, 21 males and 27 L3 juveniles in each KMW and DH1 populations (obtained from
Measurements: see Tables 1 and 2.
Morphometric data for
Characters | Female | Male | Juvenile |
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n | 20 | 20 | 27 |
Total body length | 1387 ± 63 (1307–1566) | 987 ± 89 (812–1156) | 653 ± 20 (626–704) |
a | 14.8 ± 1.4 (12.8–17.4) | 21 ± 2.7 (13.0–24.0) | 22 ± 1.7 (19.6–28.0) |
b | 7.8 ± 0.7 (6.9–10.0) | 5.9 ± 0.6 (4.7–7.0) | 4.4 ± 0.2 (4.0–5.0) |
c | 26 ± 2.2 (22.0–33.0) | 25 ± 2.7 (21.0–30.0) | 17 ± 1.2 (15.0–21.0) |
c’ | 1.8 ± 0.2 (1.5–2.3) | 1.7 ± 0.2 (1.2–2.2) | 2.0 ± 0.2 (1.4–2.6) |
V | 70 ± 2.1 (66–74) | – | – |
Lip length | 5.0 ± 0.6 (4–7) | 4.6 ± 0.8 (3–6) | 4.3 ± 0.7 (3–6) |
Lip region width | 8.9 ± 0.8 (8–11) | 6.3 ± 0.7 (5–8) | 5.0 ± 0.5 (4–5) |
Stoma length | 12.9 ± 2.2 (9–15) | 14.6 ± 1.9 (11–17) | 12 ± 1.6 (7–14) |
Pharyngeal corpus length | 108 ± 8.1 (88–124) | 93 ± 8.9 (81–108) | 82 ± 7.4 (69–99) |
Isthmus length | 37 ± 4.4 (28–46) | 35 ± 6.0 (28–50) | 29 ± 4.4 (21–37) |
Basal bulb length | 39 ± 4.9 (31–53) | 34 ± 3.4 (28–41) | 24 ± 1.5 (21–27) |
Pharynx length | 184 ± 11.1 (159–202) | 161 ± 12.9 (142–181) | 136 ± 7.8 (116–149) |
Nerve ring – ant. end | 113 ± 14 (91–151) | 106 ± 10.2 (88–129) | 94 ± 8.4 (76–109) |
Excretory pore – ant. end | 138 ± 11.0 (112–157) | 129 ± 12.4 (112–165) | 111 ± 8.9 (94–127) |
Deirid – ant. end | 155 ± 13.4 (130–178) | 128 ± 11.9 (111–156) | – |
Neck length | 191 ± 9.7 (168–208) | 181 ± 12.8 (156–203) | 162 ± 12.4 (130–181) |
Body diam. at midbody | 95 ± 10.0 (80–112) | 49 ± 7.7 (40–73) | 30 ± 2.1 (24–33) |
Ovary length | 542 ± 60 (401–652) | – | – |
Spermatheca length | 49 ± 9.8 (33–61) | – | – |
Uterus length | 280 ± 46 (211–376) | – | – |
Postvulval uterine sac length | 100 ± 11.3 (85–112) | – | – |
Vagina length | 26 ± 6.2 (17–35) | – | – |
Body diam. at vulval level | 84 ± 5.7 (74–93) | – | – |
Vulva – anterior end | 976 ± 36 (896–1046) | – | – |
Rectum length | 24 ± 5.5 (12–32) | – | 19.5 ± 2.1 (14–22) |
Body diam. at anus | 31 ± 3.4 (22–39) | 24 ± 3.3 (18–34) | 20 ± 2.7 (16–31) |
Tail length | 54 ± 4.1 (44–62) | 39 ± 3.5 (34–48) | 40 ± 2.9 (31–45) |
Phasmid to anus distance | 30 ± 3.4 (26–35) | 28 ± 3.9 (21–36) | – |
Spicules length | – | 48 ± 4.2 (41–54) | – |
Gubernaculum length | – | 25 ± 2.8 (21–30) | – |
Morphometric data for
Characters | Female | Male | Juveniles |
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n | 20 | 20 | 27 |
Total body length | 1271 ± 112 (1060–1446) | 959 ± 74 (798–1144) | 474 ± 54 (404–636) |
a | 14.1 ± 1.3 (11.5–16.6) | 20 ± 1.8 (17.1–24.0) | 21 ± 3.5 (15.3–28.0) |
b | 6.9 ± 0.4 (6.1–7.8) | 5.0 ± 0.3 (4.2–5.6) | 3.8 ± 0.36 (3.2–4.9) |
c | 27 ± 2.2 (22.0–30.0) | 27 ± 3.0 (22–34) | 13 ± 2.3 (5.7–17.6) |
c' | 1.7 ± 0.2 (1.5–2.4) | 1.6 ± 0.1 (1.3–1.8) | 2.7 ± 0.7 (2.0–6.1) |
V | 71 ± 3.6 (60–77) | – | – |
Lip length | 4.8 ± 0.7 (3–6) | 3.8 ± 0.5 (3–5) | 2.8 ± 0.5 (2–4) |
Lip region width | 7.8 ± 1.4 (5–10) | 6.3 ± 0.7 (5–8) | 5.1 ± 0.7 (4–7) |
Stoma length | 13.1 ± 1.2 (11–16) | 12.2 ± 1.6 (8–15) | 10.4 ± 1.9 (8–15) |
Pharyngeal corpus | 107 ± 9.8 (86–125) | 113 ± 6.7 (99–123) | 85 ± 8.8 (64–100) |
Isthmus | 26 ± 4.5 (16–34) | 28 ± 4.4 (21–36) | 18.2 ± 4.0 (9–27) |
Basal bulb length | 37 ± 3.6 (30–43) | 34 ± 4.1 (26–40) | 21 ± 2.9 (16–30) |
Pharynx length | 170 ± 10.3 (155–188) | 174 ± 8.4 (146–184) | 122 ± 11.9 (105–142) |
Nerve ring – ant. end | 112 ± 6.6 (98–124) | 131 ± 8.0 (114–143) | 82 ± 9.1 (65–102) |
Excretory pore – ant. end | 134 ± 12.6 (115–161) | 157 ± 12.5 (141–192) | 95 ± 11.8 (77–123) |
Deirid – ant. end | 127 ± 17.3 (95–159) | 125 ± 7.5 (105–139) | ? |
Neck length | 184 ± 10.2 (170–202) | 191 ± 8.1 (175–207) | 125 ± 9.4 (108–153) |
Body diam. at midbody | 90 ± 10.9 (70–108) | 47 ± 3.1 (40–54) | 23 ± 3.4 (18–31) |
Ovary length | 437 ± 51 (348–532) | – | – |
Spermatheca length | 50 ± 10.4 (39–68) | – | – |
Uterus length | 385 ± 85 (256–537) | – | – |
Postvulval uterine sac length | 92 ± 8.6 (73–101) | – | – |
Vagina length | 23 ± 1.83 (19–25) | – | – |
Body diam. at vulva level | 75 ± 10.8 (56–91) | – | – |
Vulva – anterior end | 904 ± 87 (749–1043) | – | – |
Rectum length | 33 ± 4.8 (22–43) | – | 13.4 ± 2.2 (10–18) |
Body diam. at anus | 28 ± 2.4 (21-32) | 23 ± 2.7 (20-28) | 14.3 ± 1.6 (11-17) |
Tail length | 48 ± 3.8 (41-54) | 36 ± 2.5 (32-40) | 38 ± 9.9 (29-80) |
Phasmid to anus distance | 27 ± 4.0 (21-37) | 21 ± 2.1 (18–25) | – |
Spicule length | – | 45 ± 2.8 (41-50) | – |
Gubernaculum length | – | 26 ± 2.0 (22-30) | – |
Female: Body is larger, 1.31 to 1.57 mm long, in the KMW population and smaller, 1.06 to 1.45 mm, in the DH1 population, more or less fusiform with a sudden narrowing behind the vulva, tapering anteriorly from mid-pharynx to lip region, fusiform, slightly arcuated ventrally and becomes open C shaped upon heat killing. Cuticle with annuli separated from each other by a narrow groove. Lateral fields with four alae limited by five longitudinal incisures ending at tail tip terminus, showing only three incisures after the phasmids. Lip region bears six inner labial papillae and four outer cephalic papillae. Lips are in pairs, with smooth margin; primary axils are “U”-shaped, usually with acute tip; secondary axils are “V”-shaped; guard processes are absent. Labial probolae is low, triangular in section, connected by tangential ridges. Amphidial apertures are pore like, oval. Oral opening is triangular leading into a narrow cephaloboid stoma bearing well-developed refringent rhabdia, cheilostom is short with bar-shaped cheilorhabdia, gymnostom is very short and stegostom is elongated with robust rhabdia. Pharynx is cephaloboid, divided in three regions: pharyngeal corpus is slightly fusiform, 2.7 to 3.1 times the isthmus length in KMW population while 3.7 to 5.4 times in case of DH1; isthmus is robust and basal bulb is spheroid with well-developed valvular apparatus. Excretory pore is located at isthmus level, at 60 to 89% of neck length, at 53 annuli; renette cells are just behind pharyngeal bulb. Hemizonid is present just anterior to the excretory pore. Deirids are present at basal bulb level, at 70 to 92% of neck length, at 48 annuli. Nerve ring surrounds the isthmus at metacorpus-isthmus junction or slightly posterior. Intestine with anterior end with thinner walls. Reproductive system is monodelphic, prodelphic: ovary well developed, with several lines of oocytes, with or without a double flexure at postvulval region; oviduct short; spermatheca well developed, 0.4 to 0.5 times longer than the body width; uterus is very long, divided in two parts only observed in young females, one distal tubular part and other proximal swollen part with thinner walls; in old females all length usually swollen containing 16 to 30 uterine eggs, 41 to 55 µm long and 24 to 35 µm wide; post-vulval uterine sac 0.7 to 0.9 times the body width; vagina is straight or slightly arcuate, 21 to 31% of body width; vulva ventral. Rectum is distinct, shorter than anal body width with three unicellular glands at its junction with the intestine. Anus is large, directed posteriorly. Tail is straight, conoid, truncated to slightly rounded terminus with 15 to 20 annuli ventrally. Phasmids are distinct pore like and located at 59 to 62% of tail length.
Male: Body is 0.81 to 1.16 mm long in the KMW population, and 0.80 to 1.14 mm long in the DH1 population, “J” shaped after heat killing with general morphology similar to female. Reproductive system is monorchic with testis ventrally reflexed anteriorly. Two deep latero-subventral grooves are extended from the sides of the cloacal apparatus approximately to the first preanal pair of the papillae. Genital papillae are in eight pairs, three pairs are pre-cloacal and five pairs are post-cloacal (two at mid tail length, one lateral at lateral field and one subventral, and three terminal, two subventral and one subdorsal), and one midventral papillae. Phasmids are well observed, located posterior to the anterior lateral papillae, at 67 to 70% of tail length. Spicules are long, broad and arcuate, larger than gubernaculum, with manubrium reduced, ventrally bent, rounded-elongate, calamus is conoid and lamina is slightly ventral curved with angular dorsal hump, long ventral velum and very thin rounded tip. Gubernaculum with manubrium-corpus is almost straight, well developed crura with acute tip. Tail is conoid, ventrally curved, with blunt terminus bearing a short fine mucro.
Third stage juvenile (L3): Body is robust, 0.62 to 0.70 mm long in the KMW population, and 0.40 to 0.64 mm in the DH1 population, elongate, straight or slightly curved at posterior end. Cuticle is almost smooth; lip region is similar to adult specimens. Stoma is narrow. Pharynx is clearly visible and differentiated into the three cephaloboid parts. Nerve ring surrounds the isthmus. Excretory pore is at isthmus level. Deirid is obscure. Cardia is reduced, surrounded by intestinal tissue. Rectum is 6 to 7% times the rectum width. Anus is prominent. Tail is conoid with an acute tip.
The material examined of
Both populations (KMW and DH1) examined now of
Comparative morphometrics of females from populations of
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Reference | Present paper | Siddiqi et al. (1992) | Khan and Hussain (1997) | Pervez (2011) | Nahiyoon et al. (2019) | Nahiyoon et al. (2019) | Steiner (1936); Siddiqi et al. (1992) | Andrássy (1967) | Bussau (1991) | Siddiqi et al. (1992) | De Ley et al. (1999) | Abolafia and Peña-Santiago (2002) | Mehdizadeh et al. (2013) | Pervez (2011) | Rashid and Heyns (1990) | De Ley et al. (1990) | De Ley et al. (1999) | Abolafia and Peña-Santiago, 2017a, b | Thorne (1925) | Akhtar (1962) |
Country | India | Pakistan | Pakistan | India | Pakistan | Pakistan | Israel | Mongolia | Denmark | Malawi | USA | Spain | Iran | India | Namibia | Senegal | USA | Namibia | USA | Pakistan |
n (♀) | 40 | 30 | 20 | 10 | 15 | 15 | 8 | 12 | 1 | 22 | 30 | 4 | 3 | 10 | 21 | 5 | 15 | 3 | 1 | 1 |
L | 1.06–1.57 | 0.88–1.21 | 0.86–1.20 | 0.99–1.19 | 0.80–1.70 | 0.8–1.70 | 0.63–0.78 | 0.67–0.77 | 0.88 | 0.64–0.91 | 0.86–1.53 | 0.70–0.91 | 0.63–0.72 | 0.61–0.68 | 0.57–0.88 | 0.45–0.67 | 0.88–1.19 | 0.74–0.84 | 1.2 | 0.8 |
A | 11.5–17.4 | 21.0–30.0 | 21.0–30.0 | 10.5–15.8 | 8.1–15.5 | 8.1–15.5 | 15.0–16.0 | 15.0–17.0 | 24.4 | 17–23 | 16.0–23.0 | 17.5–22.9 | 21.0–22.0 | 31.8–33.1 | 16.8–24.0 | 18.3–19.6 | 15.0–19.0 | 18.1–22.8 | 20.0 | 23.2 |
B | 6.1–10.0 | 4.6–5.9 | 4.86–5.8 | 4.8–6.1 | 4.5–8.0 | 4.5–8.0 | 5.0–5.6 | 4.8–5.5 | 4.7 | 4.7–5.6 | 5.6–7.8 | 4.5–5.5 | 4.2–5.1 | 4.2–4.4 | 3.5–5.3 | 3.4–4.6 | 5.7–6.9 | 4.4–5.0 | 5.7 | 6.0 |
C | 22.0–33.0 | 20.0–26.0 | 20–26 | 20.6–23 | 16.0–29.2 | 16–29.2 | 17–21 | 16.0–19.0 | 18.1 | 16.0–18.3 | 20.0–30.0 | 19.0–20.3 | 16.0–20.0 | 14.3–17.9 | 16.2–21.8 | 15.1–18.0 | 20.0–26.0 | 16.5–19.6 | 18.0 | 17.7 |
c' | 1.5–2.4 | 1.8–2.4 | 1.8–2.3 | 1.4–1.8 | 1.1–1.9 | 1.1–1.9 | ? | 1.7–2.1 | 2.1* | 2.0–2.5 | 1.4–2.1 | 1.6–2.1 | 1.8–2.1 | 1.2–1.8 | 1.4–2.3 | 2.0–2.4 | 1.4–1.9 | 1.7–2.1 | 2.0 | 2.5–2.7* |
V | 60–77 | 65–74 | ? | 68–74 | 70–75 | 70.2–75 | 69–71 | 65–71 | 69 | 67–71 | 67–71 | 64–69 | 67–70 | 77* | 64–72 | 65–70 | 84* | 66–70 | 70 | 65 |
Lip region width | 5–11 | 11–13 | 11–13 | 11–13 | 9–10 | 12–15 | 9–10 | ? | 6.7* | 5.9* | 9* | 3 | 10–11 | 8.6–12 | ? | ? | ? | 5.5* | ? | 7* |
Stoma length | 9–18 | 12* | 12–18 | 16–16 | 12–14 | 12–22 | 12–14 | ? | 20* | 14.7* | 11–16 | 11–14 | 13–14 | 8–12 | 11–14 | 8–10 | 12–15 | 10–12 | 20 | 10* |
Procorpus | 37–69 | ? | ? | ? | ? | ? | ? | ? | ? | ? | 50–79 | ? | ? | ? | ? | ? | 60–85 | ? | ? | ? |
Metacorpus | 43–80 | ? | ? | ? | ? | ? | ? | ? | ? | ? | 30–49 | ? | ? | ? | ? | ? | 35–46 | ? | ? | ? |
Isthmus | 16–46 | 21.9* | 20* | ? | 20–26 | 14–28 | 20–26 | ? | 23* | 22–36 | 20–37 | 22–25 | 21–25 | 27.6* | 14–15* | 15–24 | 17–23 | 15 –23 | ? | ? |
Bulb length | 30–53 | 29–34 | 29–34 | 24–38 | 19–21 | 26–32.5 | 19–21 | ? | 19.8* | 19–21 | 22–32 | 22–27 | 19–26 | 24* | ? | 15–21 | 22–28 | 24–25 | ? | 13–25 |
Pharynx length | 155–202 | 190–218 | 190–218 | 179–223 | 129–157 | 166–228 | 129–157 | ? | 157* | 135–178 | 205–316* | 135–166 | 133–141 | 145–154 | ? | 123 –159 | 229–313* | 137–164 | 210 | 8–14 |
Nerve ring–ant. end | 91–151 | 163* | ? | 151* | 92–114 | 125–136 | 92–114 | ? | 132* | 106* | 113–174 | 107–126 | 109–117 | 101–105 | 89–142 | 82–108 | 125–148 | 119–135 | 162 | 78* |
Excretory pore–ant. end | 112–161 | 140–188 | 140–188 | 155–174 | ? | 136–150 | ? | ? | 139* | 118–134 | 131–209 | 114–148 | 123–135 | ? | 89–157 | 83–118 | 154–195 | 127–142 | ? | 81* |
Neck region | 168–208 | 215* | 202–236* | 194–239* | ? | 178–250* | ? | ? | 177* | 137* | 157–332* | 146–180 | 139–150 | 153–166* | 120–177 | 134–153 | 241–327* | 149–172 | ? | ? |
Midbody diam. | 70–112 | 56.6* | ? | 59–113 | ? | 52–170 | ? | ? | 40* | 39* | 49–73 | 33–40 | 30–34 | 18–21 | ? | 13–17 | 49–72 | 35–41 | ? | 29.4* |
Vulva – anterior end | 749–1046 | 680–840 | 573* | 637* | 503–550 | 616–1225 | 503–550 | ? | 650* | 503–550 | 1001* | 447–629 | 436–491 | 495.2* | ? | ? | 875* | ? | 840 | 291.6* |
Body diam. at anus | 21–39 | 29* | 19* | 28–36 | ? | 25–50 | ? | ? | 30* | 18* | 22–31 | 18–22 | 19–20 | 19–21 | ? | 20–23 | 25* | 21–25 | 34 | 15* |
Rectum length | 12–43 | 37.5* | ? | 31–42 | ? | 24–30 | ? | ? | ? | 19* | 27–42 | 22–25 | 21–29 | 17.2* | ? | ? | 23–32 | 17.8 | ? | ? |
Tail length | 41–62 | 39–50 | 39–50 | 10–15 | 35–45 | 42–55 | 35–45 | ? | 50* | 39–56 | 41–60 | 36–45 | 35–40 | 37–42 | 61–111 | 30–37 | 41–52 | 88–91 | 66 | 39–43 |
Phasmid to anus distance | 21–39 | 10–19 | 23.3* | ? | ? | ? | ? | ? | ? | 18.8* | 22–36 | 16–29 | 17.5* | 20.7* | 41–66 | ? | 21–32 | 56–64 | ? | ? |
D% (EP/ES×100) | 64–101 | 74–86* | 74–86* | 82* | 79* | 79* | ? | ? | 88* | 80.5* | 64–66 | 87* | 94* | ? | ? | ? | 68* | 88* | ? | ? |
E% (EP/T×100) | 206–320 | 359–376* | 359–376* | ? | ? | 285* | ? | ? | 278* | 265.3* | 312–348 | 323* | 344* | ? | ? | ? | 378* | 150* | ? | ? |
Additionally,
Comparative morphometrics of males from populations of
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Reference | Present paper | Rashid and Heyns (1990) | De Ley et al. (1990) | De Ley et al. (1999) | Abolafia and Peña-Santiago, 2017a, b | Nahiyoon et al. (2019) | Nahiyoon et al. (2019) | Steiner (1936); Siddiqi et al. (1992) | Andrássy (1967) | Siddiqi et al. (1992) | De Ley et al. (1999) | Abolafia, Peña-Santiago (2002) | Mehdizadeh et al. (2013) | Pervez (2011) | Steiner (1935) |
Country (State) | India | Namibia | Senegal | USA | Namibia | Pakistan | Pakistan | Israel | Mongolia | Malawi | USA | Spain | Iran | India | USA |
n (♂) | 40 | 26 | 6 | 20 | 6 | 15 | 15 | 8 | 6 | 10 | 10 | 2 | 4 | 10 | 1 |
L | 0.79–1.16 | 0.53–0.94 | 0.54–0.65 | 0.70–1.03 | 0.68–0.84 | 0.71–1.39 | 0.71–1.39 | 0.63–0.71 | 0.56–0.59 | 0.56–0.87 | 0.98–1.18 | 0.69, 0.73 | 0.70–0.86 | 0.61–0.62 | 0.9 |
a | 13.0–24.0 | 16.7–26.4 | 20.2–21.7 | 16.0–26.0 | 17.8–23.7 | 11.1–16.6 | 11.1–16.6 | 14.0–16.0 | 12.0–15.0 | 22–26 | 20.0–25.0 | 24.0, 25.0 | 22.0–30.0 | 27.0–31.0 | 27.0 |
b | 4.2–7.0 | 4.0–5.9 | 4.0–4.8 | 4.3–6.8 | 4.1–5.1 | 4.6–7.5 | 4.6–7.5 | 4.9–5.1 | 4.2–4.8 | 4.3–5.4 | 6.0–7.8 | 5.0, 5.2 | 4.8–5.4 | 4.0–4.2 | 7.2 |
c | 21.0–34.0 | 14.4–21.7 | 14.9–18.0 | 17.0–23.0 | 17.3–20.3 | 19.5–27.8 | 19.5–27.8 | 15–18 | 14.0–16.0 | 15–18 | 20.0–24.0 | 18.8, 16.6 | 14.0–18.0 | 16.0–18.0 | 18.0 |
c’ | 1.2–2.2 | 1.4–2.1 | 1.6–2.0 | 1.2–1.6 | 1.2–1.5 | 0.9–1.3 | 0.9–1.3 | ? | 1.9* | 2.1* | 1.3–1.7 | 1.3, 1.7 | 1.6–2.0 | 1.4–1.8 | 1.8* |
Lip region width | 5–8 | ? | ? | ? | ? | 11–14 | 11–14 | ? | ? | ? | ? | 3, 4 | 3–4 | ? | ? |
Stoma length | 8–17 | ? | 08–10 | 11–13 | 11–12 | 12–15 | 12–15 | ? | ? | ? | 10–14 | 11, 12 | 10–11 | 11–12 | ? |
Procorpus length | 28–59 | ? | ? | 58–66 | ? | ? | ? | ? | ? | ? | ? | 11, 11 | 12–14 | ? | ? |
Metacorpus length | 45–75 | ? | ? | 32–40 | ? | ? | ? | ? | ? | ? | ? | ? | ? | ? | ? |
Isthmus length | 21–50 | ? | 15–25 | 16–23 | 19–22 | 16–20 | 16–20 | 22–23 | ? | ? | ? | ? | 17–32 | ? | ? |
Bulb length | 26–41 | ? | 16–23 | 17–27 | 22–28 | 24–30 | 24–30 | ? | ? | ? | 22–30 | 22, 23 | 20–26 | ? | ? |
Pharynx length | 142–184 | ? | 122–151* | 205–260 | 122–161 | 150–188 | 150–188 | 129–143 | ? | ? | 210–293 | 100, 119 | 131–149 | 145–152 | ? |
Nerve ring – ant. End | 88–143 | ? | 90–104 | 108–135 | 120–132 | 112–134 | 112–134 | 100–115 | ? | ? | 117–151 | 140, 140 | 110–127 | 106–110 | ? |
Excretory pore – ant end | 112–192 | ? | 78–91 | 121–175 | 119–141 | 132–144 | 132–144 | ? | ? | ? | 135–192 | 121, ? | 128–143 | ? | ? |
Neck length | 156–207 | 130–174 | 128–148 | ? | 156–168 | 162–203* | 162–203* | ? | ? | ? | 133–188 | 143–160 | ? | ? | |
Midbody diam. | 40–73 | ? | 25–31 | 39–51 | 35–42 | 64–88 | 64–88 | ? | ? | ? | 44–57 | 33, 40 | 24–40 | 19–22 | ? |
Anal body diam. | 18–34 | ? | 20–23 | 27–35 | 27–34 | 32–40 | 32–40 | ? | ? | 25* | 31–35 | 18, 22 | 24–27 | 17–24 | ? |
Tail length | 32–48 | 37–50 | 36–42 | 39–47 | 38–44 | 40–50 | 40–50 | 38–43 | ? | 54–80 | 40–58 | 22, 25 | 43–54 | 34–37 | ? |
Spicules length | 41–54 | 29–51 | 29–34 | 40–48 | 41–46 | 38–57 | 38–57 | 39–44 | 30–40 | 35–43 | 42–50 | 37 | 38–41 | 39–43 | ? |
Gubernaculum length | 21–30 | 18–35 | 14–19 | 24–30 | 26–31 | 24–35 | 24–35 | 22–28 | 20 | 19–24 | 27–34 | 23, 22 | 19–28 | 23–24 | ? |
D% (EP/ES×100) | 69–94 | ? | ? | 57–67* | 92* | 82* | 82* | ? | ? | ? | ? | 97* | ? | ? | |
E% (EP/T×100) | 242–577 | ? | ? | 310–372* | 317* | 306* | 306* | ? | ? | ? | ? | 233, 300* | 279* | ? | ? |
SW% (SL/ABD×100) | 150–294 | ? | ? | 151* | 143* | 126* | 126* | ? | ? | ? | ? | 137* | 155* | 194* | ? |
GS% (GL/SL×100) | 40–70 | ? | ? | 67* | 66* | 55* | 55* | ? | ? | ? | ? | 65* | 59* | 58* | ? |
From
From
On the other hand,
On the other hand also,
With respect to
Distance matrix analyses with other closely related populations of several
Pairwise distances of the 18 S rDNA regions between present strains of
S. No. | 18S rDNA | Country | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
1 | MK935150 |
India |
|
|
|
|
|
|
|
|
|
|
|
2 | MK935149 |
India |
|
|
|
|
|
|
|
|
|
|
|
3 | KY090631 |
Iran |
|
|
9 | 19 | 19 | 19 | 19 | 31 | 23 | 89 | |
4 | MK541681 |
Germany |
|
|
98.3 | 0 | 0 | 0 | 0 | 10 | 10 | 9 | |
5 | DQ102707 |
UK |
|
|
98.5 | 100 | 16 | 2 | 0 | 39 | 13 | 97 | |
6 | KX889085 |
South Korea |
|
|
98.5 | 100 | 99.3 | 2 | 0 | 39 | 13 | 96 | |
7 | AY284673 |
Netherlands |
|
|
98.5 | 100 | 99.9 | 99.9 | 0 | 37 | 13 | 95 | |
8 | MF325099 |
Germany |
|
|
98.5 | 100 | 100 | 100 | 100 | 19 | 4 | 86 | |
9 | AF202159 |
France |
|
|
97.4 | 98.6 | 98.3 | 98.3 | 98.4 | 98.4 | 32 | 92 | |
10 | KY119635 |
Ireland |
|
|
98.1 | 98.2 | 99.0 | 99.0 | 99.0 | 99.7 | 97.4 | 95 | |
11 | JQ237850 |
USA |
|
|
91.7 | 98.8 | 95.0 | 95.1 | 95.1 | 92.0 | 95.3 | 91.3 |
Pairwise distances of the D2D3 regions of 28 S rDNA regions between present strains of
S. No. | 28S rDNA | Country | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
1 | MK935147 |
India |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
2 | MN101167 |
India |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
3 | KY914573 |
Iran |
|
|
10 | 10 | 10 | 38 | 56 | 64 | 68 | 79 | 80 | 82 | 82 | |
4 | MF325168 |
Germany |
|
|
98.0 | 0 | 0 | 5 | 8 | 0 | 11 | 0 | 0 | 0 | 0 | |
5 | MF325157 |
Germany |
|
|
98.0 | 100 | 0 | 5 | 8 | 0 | 11 | 0 | 0 | 0 | 0 | |
6 | DQ903087 |
Germany |
|
|
94.7 | 100 | 100 | 5 | 8 | 0 | 11 | 0 | 0 | 0 | 0 | |
7 | AF147069 |
USA |
|
|
96.8 | 99.0 | 99.0 | 96.9 | 30 | 35 | 45 | 34 | 35 | 36 | 36 | |
8 | AF147067 |
USA |
|
|
95.2 | 98.5 | 98.5 | 96.1 | 97.5 | 43 | 51 | 44 | 45 | 46 | 46 | |
9 | KX889089 |
South Korea |
|
|
94.3 | 100 | 100 | 99.8 | 97.0 | 96.3 | 41 | 1 | 0 | 3 | 3 | |
10 | DQ145625 |
Belgium |
|
|
95.7 | 97.7 | 97.7 | 96.9 | 96.1 | 95.6 | 96.4 | 50 | 49 | 50 | 50 | |
11 | DQ903076 |
Sweden |
|
|
94.9 | 100 | 100 | 99.9 | 97.1 | 96.3 | 99.9 | 96.9 | 1 | 2 | 2 | |
12 | DQ903083 |
USA |
|
|
94.9 | 100 | 100 | 99.8 | 97.0 | 96.2 | 100 | 96.9 | 99.9 | 3 | 3 | |
13 | DQ145624 |
USA |
|
|
94.8 | 100 | 100 | 100 | 96.9 | 96.1 | 99.8 | 96.9 | 99.9 | 99.8 | 0 | |
14 | DQ903081 |
USA |
|
|
94.8 | 100 | 100 | 100 | 96.9 | 96.1 | 99.8 | 96.9 | 99.9 | 99.8 | 100 |
All of these data showed that
The phylogenetic analyses of the present stains based on 18 S rDNA and flanking region D2-D3 segment of 28 S rDNA gene also supported the molecular data. Phylogenetic analyses based on 18 S rDNA sequences (Fig. 5) showed a clear monophyly of the group formed by the isolates DH1 and KMW and other undescribed
Bayesian Inference tree from known and the newly sequenced
Bayesian Inference tree from known and the newly sequenced
On the other hand, Pervez (2011) described
Another species, described by Pervez (2011),
Recently, Nahiyoon et al. (2019) described a new species,