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Isolation, identification of entomopathogenic nematodes with insights into their distribution in the Syrian coast regions and virulence against Tuta absoluta

INFORMAZIONI SU QUESTO ARTICOLO

Cita

Figure 1:

Distribution map of soil sample sites and entomopathogenic nematodes in Syrian coastal regions. A: Soil sample sites. B: Sites positive for entomopathogenic nematodes. Those marked with green denote H. indica, yellow denotes H. bacteriophora, blue denotes H. pakistanense, and red denotes S. affine.
Distribution map of soil sample sites and entomopathogenic nematodes in Syrian coastal regions. A: Soil sample sites. B: Sites positive for entomopathogenic nematodes. Those marked with green denote H. indica, yellow denotes H. bacteriophora, blue denotes H. pakistanense, and red denotes S. affine.

Figure 2:

Heterorhabditis indica male. A: Male entire body. B–H: Male tail with bursal papillae. B, C: Ventral view of bursa showing papillae (3–2) in the terminal group with some variation. E, F, G, H: Lateral and ventral view of bursa showing papillae (2–2) in the terminal group with some modifications (namely, being short and swollen). I: Lateral view of spicule and gubernaculum. J: Ventral view of spicule and gubernaculum. A = 705 μm. I, J: SL = 40 μm, GL = 19 μm.
Heterorhabditis indica male. A: Male entire body. B–H: Male tail with bursal papillae. B, C: Ventral view of bursa showing papillae (3–2) in the terminal group with some variation. E, F, G, H: Lateral and ventral view of bursa showing papillae (2–2) in the terminal group with some modifications (namely, being short and swollen). I: Lateral view of spicule and gubernaculum. J: Ventral view of spicule and gubernaculum. A = 705 μm. I, J: SL = 40 μm, GL = 19 μm.

Figure 3:

Heterorhabditis bacteriophora male. A: Male entire body. B: Male tail with bursal lateral view of spicules and gubernaculum. C: Lateral view of bursa showing the arrangement of papillae, arrow refers to curved outward fourth pair. D: Lateral view of bursa showing the terminal group of bursal papillae pairs (7, 8, and 9). E: Anterior portion; the left arrows refer to the ring position and excretory pore, while the right arrow refers to the end of the esophagus. A = 861 μm. B: SL = 42 μm, GL = 20 μm. E: NR = 72 μm, ES = 101 μm.
Heterorhabditis bacteriophora male. A: Male entire body. B: Male tail with bursal lateral view of spicules and gubernaculum. C: Lateral view of bursa showing the arrangement of papillae, arrow refers to curved outward fourth pair. D: Lateral view of bursa showing the terminal group of bursal papillae pairs (7, 8, and 9). E: Anterior portion; the left arrows refer to the ring position and excretory pore, while the right arrow refers to the end of the esophagus. A = 861 μm. B: SL = 42 μm, GL = 20 μm. E: NR = 72 μm, ES = 101 μm.

Figure 4:

Heterorhabditis pakistanense male. A: Male entire body. B: The anterior portion; the left arrow refers to the nerve ring position, while the right arrow refers to the excretory pore. C, D: Male tail with bursal papilae. B, C: Ventral view of bursa showing papillae (3–2) in the terminal group with some variation in tall. D: Lateral view of bursa; the arrow refers to curved outward seventh pair. E, F, G, H: Lateral and ventral view of bursa showing (2–2) papillae in the terminal group, with some modifications (namely, being short and swollen). I: Lateral view of spicule and gubernaculm. J: Ventral view of spicule and gubernaculum. A = 705 μm. I, J: SL = 40 μm, GL = 19 μm.
Heterorhabditis pakistanense male. A: Male entire body. B: The anterior portion; the left arrow refers to the nerve ring position, while the right arrow refers to the excretory pore. C, D: Male tail with bursal papilae. B, C: Ventral view of bursa showing papillae (3–2) in the terminal group with some variation in tall. D: Lateral view of bursa; the arrow refers to curved outward seventh pair. E, F, G, H: Lateral and ventral view of bursa showing (2–2) papillae in the terminal group, with some modifications (namely, being short and swollen). I: Lateral view of spicule and gubernaculm. J: Ventral view of spicule and gubernaculum. A = 705 μm. I, J: SL = 40 μm, GL = 19 μm.

Figure 5:

Evolutionary relationship of Syrian Heterorhabditis isolates using ITS rDNA sequences. The evolutionary history was inferred by using the maximum likelihood method. The bootstrap consensus tree inferred from 1,000 replicates is taken to present the evolutionary history of the analyzed taxa. Caenorhabditis elegans was used as an outgroup.
Evolutionary relationship of Syrian Heterorhabditis isolates using ITS rDNA sequences. The evolutionary history was inferred by using the maximum likelihood method. The bootstrap consensus tree inferred from 1,000 replicates is taken to present the evolutionary history of the analyzed taxa. Caenorhabditis elegans was used as an outgroup.

Figure 6:

Cluster dendrogram of Syrian Heterorhabditis species isolates based on male morphological traits.
Cluster dendrogram of Syrian Heterorhabditis species isolates based on male morphological traits.

Figure 7:

Co-inertia loading plot for EPN species and habitat and abiotic factors data in the Syrian coastal ecosystems surveyed.
Co-inertia loading plot for EPN species and habitat and abiotic factors data in the Syrian coastal ecosystems surveyed.

Figure 8:

Larvae and pupae of T.absoluta infected by S.affine M.313. A: S. affine after completing its development within the fourth-instar larva. B: Emergence of the IJs from the infected fourth instar larva. C: Infected third instar larva. D, E: S.affine female inside pupae observed with the naked eye and 4× magnification.
Larvae and pupae of T.absoluta infected by S.affine M.313. A: S. affine after completing its development within the fourth-instar larva. B: Emergence of the IJs from the infected fourth instar larva. C: Infected third instar larva. D, E: S.affine female inside pupae observed with the naked eye and 4× magnification.

Figure 9:

Larvae and pupae of T.absoluta infected by H.indica H. A, B: infected fouth-instar larvae outside leaf and inside mine. C, D: Infected third-instar larva outside leaf, and hermaphroditic inside it. E: Infected pupae. F: Hermaphroditics inside pupa.
Larvae and pupae of T.absoluta infected by H.indica H. A, B: infected fouth-instar larvae outside leaf and inside mine. C, D: Infected third-instar larva outside leaf, and hermaphroditic inside it. E: Infected pupae. F: Hermaphroditics inside pupa.

Figure 10:

Mortality rates (mean ± SD) of third- and fourth-instar larvae inside the galleries and pupae of T.absoluta treated by LD50 inoculation rates of S.affine and H.indica (P = 0.01).
Mortality rates (mean ± SD) of third- and fourth-instar larvae inside the galleries and pupae of T.absoluta treated by LD50 inoculation rates of S.affine and H.indica (P = 0.01).

Morphometrics of male of the Syrian Heterorhabditis indica isolates. All measurements are in μm (except n, ratio, and percentage), and in the form mean ± S.D. (range).

Character SY-BS SY-E SY-AS SY-K SY-BO SY-FA SY-SH SY-HR SY-KA SY-Z SY-MS SY-ME SY-HW SY-AF H. indica Type strain Poinar et al. (1992)
n 20 20 20 20 20 20 20 20 20 20 20 20 20 20 12
L 685±89 (569–719) 697±80 (598–731) 689±75 (604–740) 715±69 (591–730) 703±81 (590–741) 718±87 (610–773) 726±89 (598–761) 741±91 (589–767) 735±85 (602–759) 749±80 (584–773) 738±90 (609–770) 745±77 (614–780) 754±94 (598–783) 762±79 (590–790) 721 (573–788)
MBD 37±2.3 (35–41) 38.5±2.4 (35–43) 37±2.1 (33–40) 41±1.5 (38–47) 40.5±1.8 (37–43) 41±1.3 (37–42) 41.5±1 (35–44) 42±1.2 (37–45) 42±1.9 (35–44) 43.5±1.6 (38–47) 43±0.9 (37–44) 44±1.4 (38–46) 43.5±1.5 (36–45) 43±1.9 (38–45) 42 (35–46)
EP 113±6.7 (105–125) 114.5±5.9 (107–121) 114±6.1 (107–127) 119±8.1 (107–127) 115.5±5.7 (109–125) 119±5.9 (105–130) 124±5.1 (109–131) 126±5.5 (110–137) 124.5±5.9 (105–134) 126±5.8 (114–136) 124.5±6.1 (107–133) 126.5±5.4 (112–136) 129±5.8 (110–139) 132±6 (109–140) 123 (109–138)
NR 71±4.5 (69–76) 72±5.8 (69–79) 71.5±4.3 (67–75) 74±4.7 (70–80) 72±6.1 (68–83) 73±5.7 (69–84) 73±4.1 (70–80) 78±5.5 (69–80) 77.5±4.3 (70–81) 78.5±4.9 (70–85) 77±3.9 (73–83) 78±4.5 (72–84) 79±5.1 (74–86) 78.5±5.2 (71–85) 75 (72–85)
ES 95±5.7 (94–104) 97.5±5.1 (92–103) 95.5±5.5 (93–105) 98±5.9 (92–107) 98.5±6.1 (94–107) 99±6.3 (95–108) 101±5.8 (95–107) 103±5.9 (94–106) 101.5±6 (96–109) 104±5.8 (94–107) 103±5.2 (96–105) 103.5±4.9 (95–106) 104.5±5.6 (94–109) 106±6 (97–109) 101 (93–109)
ABW 20±1.2 (18–22) 20±1.3 (17–21) 20.5±1.2 (18–23) 20±1.4 (17–23) 21±0.9 (18–24) 22±1.3 (19–24) 22.5±1.4 (18–24) 23±0.8 (19–24) 22±1.4 (19–25) 23±1.1 (17–25) 21.5±0.9 (19–24) 22.5±1.4 (18–25) 23.5±1.2 (18–25) 23±1.1 (18–25) 23 (19–24)
T 23±2.1 (21–26) 25±1.9 (22–29) 24.5±1.3 (23–30) 26±1.7 (21–30) 25±1.8 (22–30) 26.5±2.3 (24–33) 27.5±2.1 (24–31) 29±2.6 (21–32) 29±2 (24–31) 30±2.5 (22–32) 28±2.1 (24–31) 29.5±1.9 (24–32) 30±1.6 (24–31) 30±1.5 (25–33) 28 (24–32)
a 18.51 17.9 18.61 17.43 17.35 17.51 17.49 17.64 17.5 17.21 16.9 16.93 17.33 17.7 17.16
b 7.21 7.14 7.21 7.29 7.13 7.25 7.18 7.19 7.24 7.20 7.16 7.19 7.21 7.18 7.13
c 29.7 27.88 28.1 27.5 28.12 27 27.39 25.5 25.34 25.82 26.3 25.25 25.13 25.4 25.75
D 1.18 1.17 1.19 1.21 1.16 1.20 1.22 1.22 1.22 1.21 1.20 1.22 1.23 1.24 1.21
E 4.91 4.58 4.63 4.6 4.5 4.49 4.50 4.34 4.29 4.2 4.44 4.28 4.3 4.4 4.39
SL 37±3.9 (34–43) 38.5±3.7 (35–44) 37±3.5 (35–43) 41±4 (37–42) 39.5±3.2 (38–44) 42±2.1 (39–46) 43±3.1 (37–46) 44.5±3.4 (38–47) 43.5±3.2 (39–45) 45±3.9 (38–47) 44±3.1 (40–48) 44.5±3.2 (40–47) 46±3.8 (39–48) 47.5±4.1 (39–50) 43 (35–48)
GL 17.5±1.9 (17–22) 19±1.5 (17–22) 18±1.3 (17–21) 21±1.7 (19–22) 19.5±1.2 (18–22) 20.5±1.3 (19–23) 21±1.4 (17–22) 21.5±1.3 (19–22) 22±1.2 (19–23) 23±1.6 (19–25) 21±1.2 (20–24) 22±1.6 (19–23) 22.5±1.4 (19–24) 23±1.2 (18–24) 21 (18–22)
SW 1.85 1.92 1.80 2 1.88 1.90 1.91 1.93 1.89 1.95 2 1.97 1.95 2 1.86
GS 0.47 0.49 0.48 0.51 0.49 0.48 0.48 0.48 0.50 0.51 0.47 0.49 0.48 0.48 0.50 (0.40–0.60)

Morphometrics of male of the Syrian Heterorhabditis bacteriophora isolates. All measurements are in μm (except n, ratio, and percentage), and in the form: mean ± s.d. (range).

Character SY-ALS SY-SR SY-ALB SY-AL SY-K SY-S SY-D SY-Z SY-B H. bacteriophora Type strain Poinar (1976)
n 20 20 20 20 20 20 20 20 20 15
L 845±91 (789–910) 829±85 (778–903) 810±65 (785–895) 819±69 (791–901) 824±84 (775–915) 831±95 (793–942) 804±79 (773–891) 820±83 (786–900) 815±74 (781–906) 820 (780–960)
MBD 46.5±4.8 (37–48) 44.5±5.1 (39–47) 41±4.1 (36–43) 41±4.5 (38–46) 42.5±4.6 (37–47) 44.5±4.1 (39–46) 40±4.2 (35–45) 42±4 (36–45) 41±4.3 (36–45) 43 (38–46)
EP 126±5.5 (115–129) 124±5.7 (114–126) 119.5±5.1 (115–125) 121.5±5.5 (115–125) 123±5.9 (115–128) 124.5±5.8 (115–130) 120±5.9 (112—124) 122.5±5.1 (116–128) 122±5.4 (114–129) 121 (114–130)
NR 76±4.7 (67–79) 73±4.6 (64–77) 69±4.1 (67–74) 71±4.5 (67–77) 72±5.1 (65–80) 73.5±5.2 (66–83) 68.5±4 (65–75) 71.5±4.3 (67–76) 71±4.7 (66–78) 72 (65–81)
ES 103.5±2.1 (100–104) 103±2 (99–104.5) 102±1.7 (99–104) 101±1.1 (100–103) 102.5±1.2 (98–104) 103±1.2 (99–104) 100±0.8 (98–102) 101±0.9 (99–103) 100.5±1 (97–104) 103 (99–105)
ABW 24.5±1.7 (22–26) 23±1.3 (21–24) 21±1.1 (20–23) 21.5±1.5 (20–24) 23±1.2 (21–24) 23.5±1 (21–25) 21±0.9 (20–23) 23±1.4 (20–24) 21±1.1 (20–24) 23 (22–25)
T 32.5±3.9 (24–34) 29±4.3 (22–32) 25±3.8 (24–30) 26.5±3.1 (24–31) 27±4.1 (22–34) 29.5±3.5 (24–35) 23.5±4.5 (21–30) 27±3.4 (24–30) 26±3.1 (24–31) 28
a 18.1 18.6 19.7 19.9 19.3 18.6 20 19.5 19.8 19
b 8.16 8.04 7.94 8.10 8.03 8.06 8.04 8.11 8.01 7.9
c 25.22 28.58 32.5 30.9 30.5 28.16 24.73 30.3 31.34 29.2
D 1.21 1.20 1.17 1.20 1.2 1.20 1.2 1.21 1.21 1.17
E 3.76 4.27 4.78 4.58 4.55 4.22 5.1 4.53 4.69 4.32
SL 42.5±2.5 (37–44) 40±2.6 (35–43) 37.5±2 (36–41) 39±1.7 (37–44) 39.5±1.8 (36–44) 41±2.9 (36–46) 37±2 (35–42) 39±1.7 (36–42) 39±1.6 (35–42) 40 (36–44)
GL 21±0.8 (19–23) 19±0.7 (18–23) 18±0.7 (17–22) 19±0.9 (18–23) 20±1 (17–23) 21±1.1 (18–24) 19±1 (17–22) 20±1.2 (17–23) 19±1.1 (17–23) 20 (18–25)
SW 1.73 1.73 1.78 1.81 1.71 1.74 1.76 1.69 1.85 1.74
GS 0.49 0.47 0.48 0.48 0.48 0.51 0.51 0.51 0.48 0.50

Morphometrics of infective juvenile and male of the Syrian Heterorhabditis pakistanense isolates. All measurements are in μm (except n, ratio, and percentage), and in the form mean ± S.D. (range).

Character Heterorhabditis pakistanense SY-BH Heterorhabditis pakistanense SY-ZO Heterorhabditis pakistanense n. sp. (Shahina et al., 2016)



Stage Infective Juvenile Male Infective Juvenile Male Infective Juvenile Male
n 20 20 20 20 25 25
L 575±23 (536–597) 839.5±91 (710–945) 583±31 (542–603) 810±85 (721–991) 581 (558–624) 819 (720–1013)
MBD 20±2.7 (17–21) 39.5±2.4 (36–41) 19±0.8 (17–22) 38±2.1 (35–40) 21 (19–23) 39.8 (38–43)
EP 95.5±3.6 (85–98) 120.4±3.2 (113–129) 92±2.7 (87–98) 119±3.4 (114–125) 99.3 (95–106) 122.8 (112–133)
NR 83±2.5 (70–89) 87±3.8 (81–97) 81±1.5 (73–86) 85.6±4.2 (79–95) 82 (73–90) 89 (80–110)
ES 116.5±2.9 (109–121) 100.5±1.1 (97–103) 114±1.6 (107–119) 98.5±0.9 (97–101) 117 (113–125) 102 (100–105)
ABW 12.5±1.5 (9.4–15) 23.3±0.9 (21–25) 12±1.2 (10–15) 22.5±1.4 (21–26) 13.7 (12–16) 24.5 (22–26)
T 96.5±2.3 (92–103) 38±1.8 (32–41) 94±2.1 (90–105) 35.5±2.3 (31–40) 99 (95–110) 37 (30–42)
a 28.75 21.2 30.6 21.3 27.4 (25–29) 20.5 (18–23.5)
b 4.9 8.35 5.11 8.22 4.8 (4.7–5.3) 7.9 (7.2–9.8)
c 5.95 22 6.2 22.8 5.78 (5.4–6.2) 22 (19.3–25)
D 0.81 1.19 0.80 1.20 0.84 (0.78–0.97) 1.19 (1.1–1.26)
E 0.98 3.16 0.97 3.35 1 (0.95–1.07) 3.31 (2.67–4)
SL - 39±2.4 (35–41) - 37±2.7 (34–40) - 38.5 (35–42)
GL - 21.4±1.9 (19–23) - 20.5±1.5 (19–22) - 21 (20–22)
SW - 1.67 - 1.64 - 1.56 (1.44–1.91)
GS - 0.54 - 0.55 - 0.58 (0.48–0.65)

Locality, coordinates, altitude, sampling date, habitat, and soil characteristics of positive samples.

Species Site/Location GenBank accession no. Coordinates Altitude Sampling date Habitats Soil types pH Organic matter %
H. indica Zeghreen / Latakia OP221993 35°43′39″N 35°52′39″E 40 m Autumn Olive field Sandy 7.3 3.39
H. indica Al-Hinadi / Latakia OP221988 35°51′48″N 35°30′12″E 36 m Autumn Citrus orchard Silty loam 7.5 1.4
H. indica Ein-Albida / Latakia OP221987 35°39′26″N 35°35′18″E 207 m Autumn Citrus orchard Silty loam 7.5 5.19
H. indica Al-Meherfha/Latakia OP221992 35°36′12″N 35°51′20″E 214 m Autumn Olive orchard Sandy loam 6.1 2.3
H. indica Al-Shamiea /Latakia OP221989 35°38′22″N 35°48′30″E 61 m Autumn Citrus orchard Silty loam 6.5 3.72
H. indica Fattiro /Latakia OP221991 35°37′19″N 35°15′24″E 138 m Autumn Olive orchard Clay 6.2 2
H. indica Kersana/Latakia OP221983 35°37′18″N 35°49′40″E 53 m Autumn Olive orchard Sandy loam 6.0 5.72
H. indica Al-Basa/Latakia OP221984 35°30′14″N 35°52′15″E 46 m Autumn Vegetables Silty loam 6.5 2.33
H. indica Al-Borjan /Latakia OP221986 35°17′34″N 35°58′42″E 47 m Autumn Vegetables Sandy loam 6.1 3.66
H. indica Al-Meghreet /Latakia OP221990 35°63′17″N 35°49′53″E 81 m Spring Citrus orchard Silty loam 6.4 2.9
H. indica Al-Hwiz/Latakia OP221996 35°20′27″N 36°00′02″E 108 m Spring Grassland Sandy loam 5.2 2.26
H. indica Hraeson / Tartus OP221985 35°15′01″N 35°57′01″E 15 m Spring Citrus orchard Silty loam 8.1 2.73
H. indica Fneiteq /Tartus OP221994 35°06′51″N 36°07′17″E 597 m Autumn Walnut orchard Silty loam 8.1 1.73
H. indica Karm-Alten/Tartus OP221995 35°07′01″N 36°06′20″E 580 m Spring Grassland Sandy 7.5 3.02
H. bacteriophora Al-Snobr /Latakia OP222916 35°28′45″N 35°53′09″E 26 m Spring Grassland Clay loam 5.8 6.2
H. bacteriophora Zeghreen /Latakia OP222020 35°43′45″N 35°52′57″E 46 m Autumn Citrus orchard Clay loam 6.7 4.72
H. bacteriophora Al-Shelfatea/Latakia OP222019 35°32′54″N 35°53′53″E 41 m Autumn Citrus orchard Silty loam 7.1 3.3
H. bacteriophora Al-Serskiea/Latakia OP222018 35°42′33″N 35°55′36″E 85 m Autumn Vegetables Sandy loam 6.7 5.39
H. bacteriophora Ras-Alein /Latakia OP222015 35°17′38″N 35°57′53″E 32 m Spring Tobacco field Silty loam 6.0 1.46
H. bacteriophora Al-Basa /Latakia OP222017 35°29′34″N 35°50′43″E 4 m Autumn Citrus orchard Silty loam 7.7 2.67
H. bacteriophora Dimsarkho /Latakia OP222012 35°33′07″N 35°46′26″E 15 m Autumn Tobacco field Silty loam 5.9 3.06
H. bacteriophora AL-Kawkae/Tartus OP222014 35°09′24″N 36°06′47″E 486 m Spring Grassland Clay loam 6.3 2.06
H. bacteriophora Bet-shofan /Tartus OP222013 34°45′35″N 36°00′44″E 70 m Spring Citrus orchard Silty loam 7.8 1.25
H. pakistanense Al-Zobar /Latakia OP235514 35°37′57″N 36°00′12″E 213 m Autumn Pear orchard Sandy loam 6.8 1.86
H. pakistanense Al-Bahloliea/Latakia OP235513 35°38′09″N 35°57′40″E 179 m Summer Vineyard Sandy loam 7.1 1.3
S. affine Amodeyha/Tartus OM350119 35°06′10″N 36°07′57″E 759 m Spring Walnut orchard Silty loam 6.1 6
S. affine Fneitiq/Tartus OL437016 35°06′02″N 36°08′51″E 604 m Winter Grass land Silty 6.4 4.1

Vegetation habitat and abiotic factors with their corresponding codes.

Habitat Code-Hab Soil characteristics Code Altitude and Season Code
Citrus orchard Hab1 Soil type

C

Sa

Si

SaL

SiL

CL

Altitude

Alt1

Alt2

Alt3

Alt4

Olive orchard Hab2

Clay

Sand

Silty

Sandy loam

Silty loam

Clay loam

0–200 m

200–400 m

400–600 m

600–800 m

Grassland Hab3
Vegetables Hab4 Season
Walnut orchard Hab5

pH

<6.5

= 6.5–7.5

>7.5

pH1

pH2

pH3

Autumn

Spring

Summer

Winter

S1

S2

S3

S4

Tobacco field Hab6
Pear orchard Hab7

Organic matter content

1–2%

2–4%

4–6%

OM1

OM2

OM3

Vineyard Hab8

Morphometrics of infective juvenile of the Syrian Heterorhabditis bacteriophora isolates. All measurements are in μm (except n, ratio, and percentage), and in the form mean ± S.D. (range).

Character SY-ALS SY-SR SY-ALB SY-AL SY-K SY-S SY-D SY-Z SY-B H. bacteriophora Type strain Poinar (1976)
n 20 20 20 20 20 20 20 20 20 15
L 603±80 (532–613) 600±67 (521–611) 597±71 (527–608) 589±67 (529–603) 593±78 (522–610) 578±79 (524–602) 553±51 (519–597) 564±63 (520–602) 575±59 (518–590) 588 (512–617)
MBD 26±3.2 (18–33) 26±3 (17–31) 24.5±2.8 (18–29) 24±2.9 (18–31) 24±2.9 (18–33) 23.5±2.8 (18–31) 21.5±3.1 (18–29) 23±3.3 (18–31) 23±2.7 (18–31) 23 (18–31)
EP 106.5±5.4 (91–112) 106±5.6 (86–114) 105.5±6 (89–109) 102±6.3 (85–105) 104±6.8 (89–110) 98.5±5.7 (87–102) 93.5±5.1 (88–100) 94.5±5 (89–101) 97±5.5 (88–100) 103 (87–110)
NR 90±4.8 (75–95) 88.5±4.5 (72–92) 88.5±4.6 (72–90) 86.5±4.9 (74–90) 88±4.7 (71–94) 84±5 (72–90) 80.5±4.3 (70–91) 84±4.6 (71–92) 83±4.2 (70–89) 85 (72–93)
ES 132±6.2 (113–138) 131.5±5.7 (103–135) 129.5±5.9 (102–135) 126.5±6.1 (105–130) 128±6.3 (104–138) 124±6 (102–132) 114±5.9 (104–130) 119±5.6 (107–130) 122±6.2 (106–129) 125 (100–139)
ABW 8.2±1.2 (6–8) 8±1.1 (6.3–8.4) 7.8±0.9 (6.5–8) 7.4±0.7 (6.5–7.9) 7.5±1 (6.5–8) 7.2±0.7 (6.3–7.9) 6.5±0.9 (6.2–7.4) 6.8±0.9 (6.5–7.8) 7±1 (6–7.4) -
T 108.5±6.7 (89–115) 107±6.4 (81–114) 104±6.1 (83–109) 100±5.8 (85–104) 102±6.2 (81–112) 95±6.3 (82–101) 89±6 (81–100) 92.5±5.6 (82–102) 93.5±5.1 (81–98) 98 (83–112)
a 23.1 23 24.36 24.5 24.7 24.59 25.72 24.52 25 25 (17–30)
b 4.56 4.56 4.61 4.65 4.63 4.66 4.85 4.73 4.71 4.5 (4–5.1)
c 5.55 5.60 5.74 5.89 5.81 6 6.21 6 6.14 6.2 (5.5–7)
D 0.80 0.80 0.81 0.80 0.83 0.79 0.82 0.79 0.79 0.84 (0.76–0.92)
E 0.98 0.99 1.01 1.02 1 1.03 1.05 1.02 1.03 1.12 (1.03–1.30)

Mortality of third- and fourth-instar larvae of Tuta absoluta on outside leaves at each dose rate for three different entomopathogenic nematodes (mean ± S.D.).

EPNs species Dose 1IJ/ L Dose 5Ij/ L Dose 10Ij/L Dose 15Ij/ L Dose 25Ij/ L Dose 50Ij/ L
Third-instar larvae of T. absoluta S. affine. 313 25.00a±1.62 37.50a±2.16 45.83a±1.33 54.17a±1.45 66.67a±0 79.17a±0.95
H. indica. Fn 16.67b±0 26.17ab±2.27 37.50a±1.33 45.83a±1.45 62.50a±1.43 70.83a±0.84
H. bacteriophora. H 8.33bc±0.32 16.67bc±0.15 25.00b±2.62 33.33b±0 37.50b±1.43 45.83b±0.84
Control 0c±0 0c±0 0c±0 0c±0 0c±0 0c±0
P=0.00139** P=0.0000657*** P=0.0000114*** P=0.0000105*** P=0.0000559*** P=0.0000011***
df= 3 df= 3 df=3 df=3 df=3 df=3
F=10 F=30.7 F=27.6 F= 65.33 F=108.5 F=97.22

Fourth-instar larvae of T. absoluta S. affine. 313 29.17a±1.33 41.67a±1.64 50.0a ±0 59.52a±1.16 71.43a±1.59 91.67a±1.62
H. indica. Fn 20.83ab±1.33 36.50ab±1.33 41.67ab±1.62 50.0ab±0 50.00a±1.45 87.50a±1.33
H. bacteriophora. H 12.50ab±2.17 25.00b±1.64 33.33b±0 41.67b±1.62 45.83a±1.33 54.17b±1.95
Control 4.17b±1.62 4.17c±1.34 4.17c±1.33 4.17c±1.33 4.17b±1.33 4.17c±1.33
P=0.00671** P=0.000318*** P=0.0000175*** P=0.0000462*** P=0.00295** P=0.0000311***
df= 3 df= 3 df= 3 df= 3 df= 3 df= 3
F=6.67 F=14 F=39.29 F=32.78 F=8.29 F=53.86

Distribution of EPNs in Syrian coast region according to the vegetation habitat, altitude and sampling seasons variables.

Habitat Frequency Abundance No. of identified samples to species
Coniferous forest 0 c 0 b 0
Broadleaf forest 0 c 0 b 0
Citrus orchards 27.27 a 2.4 a 9
Olive orchards 10.81b 1.06 a 4
Walnut orchards 8 b 0.53 a 2
Pear orchards 4.34 b,c 0.26 a 1
vineyard 4 bc 0.26 a 1
Fruit orchards (e.g., cherry, apple) 0 c 0 b 0
Vegetable fields 7.5 b 0.8 a 3
Oil crop 0 c 0 b 0
Leguminous crop 0 c 0 b 0
Leafy vegetables 0 c 0 b 0
Tobacco fields 8.33b 0.53 a 2
Riverbanks 0 c 0 b 0
Greenhouses 0 c 0 b 0
Fodder crop 0 c 0 b 0
Grassland 11.11b 1.33 a 5
Medicinal and aromatic plants 0 c 0 b 0
Sea coast 0 c 0 b 0
Public parks 0 c 0 b 0
P = 0.01491 P = 0.01892
Altitude
0–200 8.37a 19.31 18
200–400 1.79b 3.79 3
400–600 1.93b 3.94 3
600–800 1.39b 2.7 2
>800 0c 0 0
P = 0.0138 P = 0.1565
Season
Autumn 5.01 a 10.37 16
Spring 2.4a,b 6.33 9
Summer 1.4b 1.96 1
Winter 1.78 b 2.12 1
P = 0.0474 P = 0.08053

Morphometrics of infective juvenile of Syrian Heterorhabditis indica isolates. All measurements are in μm (except n, ratio, and percentage), and in the form: mean ± s.d. (range).

Character SY-BS SY-E SY-AS SY-K SY-BO SY-FA SY-SH SY-HR SY-KA SY-Z SY-MS SY-ME SY-HW SY-AF H. indica Type strain Poinar et al. (1992)
n 20 20 20 20 20 20 20 20 20 20 20 20 20 20 25
L 532±31 (503–589) 551±25 (532–591) 547±45 (523–598) 559±37 (518–589) 553±37 (510–595) 561±41 (529–602) 569±27 (520–596) 578±46 (545–610) 570±21 (531–604) 589±69 (520–610) 597±51 (532–614) 602±62 (541–610) 607±66 (539–615) 604±59 (551–618) 528 (479–573)
MBD 20.5±0.7 (18–22) 20±1.1 (19–23) 19±0.8 (17–21) 19.5±0.9 (18–22) 20 ±1 (18–21) 21±1.3 (17–24) 21±1.2 (18–23) 22±1.4 (18–24) 21.5±1.5 (19–24) 21±1.2 (18–25) 22±1.4 (17–24) 21±1.8 (18–26) 22±1.2 (19–25) 22±1 (18–25) 20 (19–22)
EP 97.2±6 (85–103) 95±5.5 (83–101) 94.5±5.3 (85–100) 96.5±5 (87–102) 94±4.9 (82–102) 96±5.2 (83–102) 101±6.1 (85–107) 98.5±4.7 (87–103) 101±4.5 (81–105) 102±6.7 (89–107) 102±6.4 (84–106) 101.5±58 (89–110) 103±6.5 (87–112) 102±6.3 (89–109) 98 (88–107)
NR 81.4±3 (70–86) 78.6±2.4 (74–87) 83±1.9 (78–88) 82.5±2.1 (73–86) 82±2.3 (75–84) 81±2.8 (70–85) 83±2.7 (74–88) 80±2.3 (75–86) 82.5±2.2 (73–85) 84±2.6 (74–88) 83.5±2.9 (71–87) 84±2.5 (75–88) 81±2.7 (72–86) 80.5±2.9 (75–84) 82 (72–85)
ES 118±4 (112–122) 117±5.1 (104–123) 118±6 (109–121) 117.5±5 (110–120) 115±5.6 (109–121) 121±5.4 (110–123) 121±5.8 (112–124) 122±7.4 (105–125) 118±6.1 (107–120) 119.5±6 (107–122) 120±6.1 (114–125) 119±5.4 (108–121) 118±5.7 (110–123) 120±5.5 (109–122) 117 (109–123)
ABW 9.6±1.1 (8–11) 7±0.7 (6.5–8) 7.1±0.8 (6.5–7.8) 7±0.7 (6.4–7.5) 6.7±1 (6.2–7.5) 7.2±0.8 (6.5–7.9) 7.5±0.9 (6.7–7.9) 7.2±0.4 (6.8–7.5) 7±0.6 (6.4–7.6) 7.8±0.9 (6.9–8) 6.8±0.8 (6.4–7.8) 6.7±0.9 (6.5–7) 7.1±0.5 (6.7–7.4) 7±0.5 (6.5–7.6) -
T 103±5.1 (88–105) 94±2.3 (90–97) 96±4.1 (92–106) 97.5±2.3 (93–102) 97±1.4 (92–99) 98.5±2.9 (89–103) 95±2.1 (90–101) 102±5 (89–107) 98±4.9 (88–105) 101±4.1 (90–110) 98.5±4.5 (93–104) 102±4 (94–107) 104±4.3 (93–105) 101±4.9 (95–107) 101 (93–109)
a 25.9 27.5 28.7 28.6 27.6 26.7 27 26.2 26.5 28 27.1 27.3 27.5 27.4 26 (25–27)
b 4.50 4.7 4.63 4.75 4.82 4.63 4.70 4.81 4.83 4.90 4.97 5 5.14 5 4.5 (4.3–4.8)
c 5.1 5.86 5.69 5.73 5.70 5.69 5.98 5.66 5.81 5.83 6 5.90 5.83 5.98 5.3 (4.5–5.6)
D 0.82 0.81 0.80 0.82 0.82 0.79 0.83 0.80 0.85 0.85 0.85 0.85 0.87 0.85 0.84 (0.79–0.90)
E 0.94 1.01 0.98 0.98 0.96 0.97 1.06 0.96 1.03 1 1.03 0.99 0.99 1 0.90 (0.83–1.03)
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