Researchs on the composition of solitary wasps of the family sphecidae (hymenoptera: apoidea) in several provinces of the northwest part of vietnam and several biological, ecological and behavioral characteristics of the mud dauber wasp sceliphron madrasp

uals number of specimens collected by the number total of all specimens collected: Number of specimens collected Rate (%) = x 100 The number total of all specimens collected 8 2.3.1.2. Methods of pinning specimens Specimens were pinned and dried and then kept in insect boxes and deposited at the Institute of Ecology and Biological Resources, Vietnam Academy of Science and technology, 18 Hoang Quoc Viet, Cau Giay, Ha Noi. 2.3.1.3. Methods of dertermination We used microscopes Olympus model SZ60 to observe morphological characters of specimens and publications with keys to determine the scientific name of species, such as: Kirby (1798), Bingham (1897), William (1919), Menke (1961) [100], Tsuneki (1963), Menke and Willink (1964), van der Vecht and van Breugel (1968), Bohart and Menke (1976), Krombein (1984), Hensen (1987, 1988, 1991), Ohl and Höhn (2011), Danilov (2012, 2015, 2018), Dollfuss (2008, 2013, 2016), Kim et al. (2014), Kumar et al. (2014) and Dörfel and Ohl (2015). Terminology used in the thesis followed that of Bohart and Menke (1976). Determination of subspecies of S. madraspatanum was based on publications of van der Vecht and van Breugel (1968) and Bohart and Menke (1976). The key was based on that of van der Vecht and van Breugel (1968). 2.3.1.4. Methods of study on the distribution of sphecid wasps of the family Sphecidae + At areas of the study, 4 habitats choosen were natural forest habitat, artificial forest and orchard habitats, shrub habitat, and resident habitat. 2.3.2. Methods of study on biological characteristics of S. madraspatanum Under the laboraroty condition, the average temperature 28,1 – 28,6C and the average humidity 80,9 – 81,2%. Research methods on biological characteristics of S. madraspatanum were based on those of Krombein (1984), Polidori et al. (2005) and Gess và Gess (2014). 2.3.2.1. Methods of study on the nesting construction New nests of S. madraspatanum collected in the field were splited to take nesting contents including: Nesting materials, nesting colours, shape of cells and nests, number of cells and number of nesting floors. Cell and nest sizes were measured by mini meter rulers. Nests of S. madrapatanum were distinguished from those of species of the genus Sceliphron by having adults that were emerged or pupae or and adults that had been died into the nest. S. madrapatanum and its subspecies were determined based on the key of van der Vecht and van Breugel (1968). 2.3.2.2. research methods of the developmental time of stages Stages of S. madraspatanum consisted of: egg, larva, prepupa, pupa and adult. We used microscope model SZ60 to observe and to describe morphological characterictics of these stages. The size of egg stage was measured by a magnifier attached with a mini meter ruler, and that of the others was measured by a mini meter ruler. Prepupae and pupae were romoved from their cocoons. The length was measured from the head to the end part of abdoment. The width of the egg and larva stages was taken at the center of the body, that of the prepupa stage was hold at the widest ternite, and that of the pupa and adult stage was measured between tegulars on the mesoscutum. Pictures were taken using Canon model SD3500 IS. Newly built nests of S. madraspatanum in the field were spilted to take nesting contents such as: eggs, larvaecocoons, adults, and prey. Eggs were collected from nesting cells provisioned in the same day. The contents were then put in small glass tubes (5 - 7 cm long and 0,6 - 1 cm in diameter). Because the immature stages of the wasp in the field is into mud nests without light, the contents were also put in the same condition (into carton boxes). Each of them was attached to a small ticket (5 cm long and 3 cm wide) that aimed to write down information of results. These contents were observed every day. Tubes contained eggs and larvae were looked at with the microscope of the magnification of 40 times. In each of these stages, 20 – 25 individuals were kept into Ethanol 95% for study on morphology.. 9 After emerging, adults of S. madraspatanum were reared into insect net cages measured by 50 x 50 x 50 cm and food of bee’s honey diluted to 50% were provided to them. Wet mud and prey were also put into theses cages. We wrote down notes related to completion time of a nesting cell and methods of provision and them dissected these cells to find eggs newly laid. These eggs would be foundation to determine time before laying of females of the wasp. Another method to determine this time was that we dissected adult females after of 4 – 7 days of age to find their eggs. The eggs with more darker colour than were looked at to know whether or not they could hatch. If the eggs laid, we would determine time before laying of the females (referrced from Ohl và Linde, 2003). Temperature and humidity in the laboratory condition were taken using Max-Min Thermo Hygro, CT-138B. 2.3.2.3. Methods of study on life time of adults Adults emerged in the laboratory were reared into plastic jars (20 cm high and 12 cm in diameter) with covers made of mosquito nets. Each jar kept 2 adults either males or females. The adults were fed with 50%-diluted bee’s honey. A piece of cotton was wetted by the honey and hang under covers. The sex was distinghuished by: normal observations and observations under magnifiers. 2.3.2.4. Mothods of study on life time of immature stages Methods used here were the same mentioned above (item 2.3.2.2). Formulas of life rates of immature stages were: The number total of eggs laid Life rate of egg (%) = x 100 The number total of eggs tested The number total of larvae emerged Life rate of larva (%) = x 100 The number total of larvae tested The number total of pupae emerged Life rate of pupa (%) = x 100 The number total of pupae tested For old nests of S. madraspatanum, each emerged hole determined as successful emergence of an adult because the holes were made by adults getting arise. 2.3.2.5. Methods of study on sex rate Newly built nests of the wasp collected in the field were choosen and numberred (from 1 to the end), then put into plastic jars (the same size mentioned above). These jars were coverred by mosquito nets and were observed every day. When adults emerged, we noted number marked on the surface of their nest and determined their sexes. The sex rate in the field was determined by catching adults and then find their sexes. We used insect nets to catch these adults. In the laboratory, we determined the sexes of adults immediately after they emerging. 2.3.3. Mothods of study on ecological characteristics of S. madraspatanum 2.3.3.1. Mothods of study on nesing sites New and old nests of S. madraspatanum were collected at all areas of study in the field. These nests were numberred, noted on time of collection, location, substrate of nests attached and highlight of nesting sites. Pictures of nesting sites were taken. In the laboratory, we analysed and assembled nesting sites of the species. 10 2.3.3.2. Methods of study on overwintering time New nests of S. madraspatanum collected from October to November were splited to take cocoons newly completed. The cocoons were put into glass tubes (50 - 80 mm long, 6 - 10 mm in diameter) and these tubes then put in plastic boxes coverred. The boxes were put under the eaves (the same natural condition). We checked the tubes once a week, and when we found pupae we determined that time was end of overwintering time. We also surveyed nests of the wasp in the field at the beginning of summer to remark the beginning time of occurrence of the species. 2.3.3.3. Methods of study on yearly occurred time of adults We investigated to find new nests of S. madraspatanum from October to November between and remarked these nests as well as ending time of action of adults in year. Sites serveyed were under bridges and desolated houses. The beginning of following April, we observed these nests, if adults emerged or emerging holes appeared we calculated the time of occurrence of the wasp in the field. Cocoons methoded as mentioned in the item of the overwintering time previously. When adults emerged we calculated the time of occurrence of the wasp in year. 2.3.3.4. Methods of study on the effect of food to life time of adults We did this study within 4 formulas: no food, fresh water, 50% diluted bee’s honey, and fruit suger cane. After immediately emerging, adults were reared into plastic jars (20 cm high and 12 cm in diameter), each with 2 adults. The jars were coverred by morquitor nets. We checked the jars every day until adults dead. 2.3.3.5. Mmethods of study on prey We chose 10 nests of S. madraspatanum collected in several different locations of the areas of study and splited them to take their contents, mainly prey. Prey taken were put into jars contained ethanol 70%. Prey were then sent to specialists to determine them to genera or species level. Occuring rate of prey into nests was showed such as: The number total of nests contained prey Occuring rate (%) = x 100 The number total of nests tested 2.3.3.6. Methods of study on predators Predators of S. madraspatanum were defined by: Observations in the field and analysis of nests of the wasp in the laboratory. 2.3.3.7. Methods of study on the effect of human activities to nesting sites of S. madraspatanum We collected information about the effect of human activities to nesting sites of S. madraspatanum through interviews, specially checked distubances of human activities to nests built into houses. Information consisted of: driving away, destroying nest, destroying nesting site, and cultivating activity. 2.3.4. Methods of study on habits of S. madraspatanum 2.3.4.1. Methods of study on nesting, egg-laying, and mating habits We chose nesting sites, which many adults of the species appeared, such as: under bridges and into desolated houses, because it would be easy for us to observe its habits. Sites chosen here for observations on nesting habits were into a desolated house built in a fruit garden at Thanh Tan commune, Luong Son district, Hoa Binh province; a desolated house at Lien Mac commune, Bac Tu Liem district, Ha Noi city; into a desolated house at Hong Minh commune, Hung Ha district, Thai Binh province. Sites chosen for observations on mating 11 habits were into a desolated house at Hong Minh commune, Hung Ha district, Thai Binh province and under bridge at Co Nhue precinct, Bac Tu Liem district, Ha Noi city. When females of the wasp began to build its nest, we observed them, wrote down notes on methods of making pieces of mud, bringing mud, and building nest with mud... and took pictures using Canon model SD 3500IS to aim analysis more than data. Nest-building time was taken by a Stopwatch model SPT-20. With cells that have not yet completed in the day, we used a violet inked pen to remark onto those cells and took a continuingobservations in the following day. Methods of study on egg-laying habits of the wasp were based on analysis of nests in which cells contained eggs and prey into. 2.3.4.2. methods of study on feeding habits of larva Feeding and cocooning habits of larvae were observed based on methods of study on the developmental time of immature stages (also see the item 2.3.2.2). 2.3.4.3. Methods of study on emerging habits of adult In the laboratory: Fresh nests of S. madraspatanum collected in the field were put plastic jars (12 cm high, 10 cm in diameter) with covers of morquito nets. The nests were observed every day, if when high-intensitied buzzes were made by adults, we observed and wrote down notes on emerging habits of the species. In the field: Fresh nests of S. madraspatanum built under a bridge at Co Nhue precinct, Bac Tu Liem district, Ha Noi city were observed. To take the emerging time of adults, we had based on the nest-built time and marking nest cells. 2.3.5. Methods of study on analysing data Data yielded in the project were analysed using the Data Analysis in Microsoft Office Excel 2007, and ANOVA. CHAPTER 3. RESULTS AND DISCUSSION 3.1. The composition of solitary wasps of the family Sphecidae and their distribution in several provinces of the northwest part of Vietnam 3.1.1. The composition of solitary wasps of the family Sphecidae The composition of solitary wasps of the family in several provinces of the northwest part of Vietnam was studied and showed in table 3.1. A total of 25 species and 6 subspecies belonging to 7 genera into 4 subfamilies were recorded. Of which, C. tanvinhensis Pham and Ohl, 2019 had decribed already, 4 species and 3 subspecies were recorded for the sphecid wasp fauna of Vietnam, and 9 species and 6 subspecies were recorded for the first time from in northwest Vietnam. Table 3.1. The composition of solitary wasps of the family Sphecidae in several provinces of the northwest part of Vietnam No. Species name Rate of collection (% ) A Subfamily Ammophilinae I Genus Ammophila 1 Ammophila clavus (Fabricius, 1775) 15.87 2 Ammophila globifrontalis Li và Yang, 1995** 0.55 3 Ammophila laevigata Smith,1856 5.47 B Subfamily Chloriontinae II Genus Chlorion 4 Chlorion lobatum (Fabricius, 1775) 2.55 12 C Subfamily Sceliphrinae III Genus Chalybion 5 Chalybion bengalense (Dahlbom, 1845) 12.59 6 Chalybion dolichothorax (Kohl, 1918) 0.18 7 Chalybion gracile Hensen, 1988*** 0.90 8 Chalybion japonicum (Gribodo, 1883)*** 9.30 9 Chalybion malignum (Kohl, 1906)** 6.02 10 Chalybion tanvinhensis Pham và Ohl, 2019* 0.54 IV Genus Sceliphron 11 Sceliphron deforme (Smith, 1856) 4.56 12.1 Sceliphron javanum petiolare Kohl, 1918*** 1.28 12.2 Sceliphron javanum chinensis van Breugel, 1968*** 0.36 13 Sceliphron madraspatanum Fabricius, 1781 20.43 13.1 Sceliphron madraspatanum sutteri van der Vecht, 1957** 0.54 13.2 Sceliphron madraspatanum andamanicum Kohl, 1918** 0.54 13.3 Sceliphron madraspatanum conspicillatum (Costa, 1864)** 3.28 13.4 Sceliphron madraspatanum kohli Sickmann, 1894*** 1.82 D Subfamily Sphecinae V Genus Isodontia 14 Isodontia aurifrons (Smith, 1859)*** 0,90 15 Isodontia chrysorrhoea (Kohl, 1890)*** 0,18 16 Isodontia elsei Hensen, 1991** 0,36 17 Isodontia nigella (Smith, 1856)** 2,37 18 Isodontia sp.1 0,18 19 Isodontia sp.2 0,90 20 Isodontia sp.3 1,82 VI Genus Prionyx 21 Prionyx viduatus (Christ, 1791)*** 1,64 VII Genus Sphex 22 Sphex argentutus Fabricius, 1787 1,46 23 Sphex diabonicus Smith, 1858 0,36 24 Sphex sericeus (Fabricius, 1804) 0,54 25 Sphex subtruncatus Dahlbom, 1843 2,19 Total 100 Remark: *new species, **new record for Vietnam, ***New record in northwest Vietnam 3.1.2. Morphological characteristics of Chalybion tanvinhensis Pham and Ohl, 2019 Specimens examined: 1♀, Tan Vinh, Luong Son, Hoa Binh, 2.v.2017, Phong Huy Pham; 2♂, Ban Chieu, Muong Thai, Phu Yen, Son La, 11.viii.2017, Phong Huy Pham. Morphology: Female (Figs. 1‒6): Body length 20 mm, forewing length 15 mm. Head (Fig. 1): In frontal view subcircular, about 1.23 times as wide as high. Mandible with inner subapical tooth. Clypeus moderately convex, without median carina; Anterior margin of clypeus with five distinct teeth: lateral tooth, separated distantly from submedian tooth, narrow and sharp; submedian tooth blunt, broader than median and lateral teeth, and as long as median tooth. Frontal line present. Ocelli not reduced; distance between hindocelli about equal to that from hindocellus to inner eye margin and about 0.43 times that from hindocellus to vertex margin. Clypeus coarsely, sparsely punctate. Frons coarsely, irregularly 13 transversely punctatorugose. Vertex and gena finely, sparsely punctate. White setae sparse on clypeus, denser than on gena. Lower inner orbit area with moderately dense pubescence. Flagellar ratio (length of the first flagellomere divided by that of the second flagellomere) 0.82; orbital ratio (shortest interocular width across vertex divided by shortest interocular width across clypeus) 0.83; clypeal ratio (length of clypeus divided by shortest interocular width across clypeus) 0.82; hypostomal cavity ratio (length of hypostomal cavity divided by width) 0.89. Mesosoma (Figs. 2‒5): In dorsal view, width between tegulae equal to that of pronotum at midlength. Pronotum with notch at middle and series of transverse striae anteriorly. mesoscutum moderately transversely striate, interspaces shallowly punctate; mesopleuron coarsely, densely punctate. Scutellum, metanotum, and upper metapleural area rather coarsely, densely punctate. Lower metapleuron conspicuously, transversely striate. Dorsal propodeum coarsely, transversely striate; side of propodeum punctate-reticulate. Mesosoma ratio (length of mesosoma divided by height) 2.18. Metasoma (Figs. 5‒6): Integument smooth, shiny, with silvery pubescence. Petiole conspicuously curved. Metasomal sternum IV without micropubescence. Petiole much shorter than hindbasitarsus. Petiole-basitarsal ratio (length of petiole divided by that of hindbasitarsus) 0.70. Color: Integument dark blue. Mid and hind legs, propodeum and metasoma with violaceous reflections; antenna black except scape and pedicel dark blue. Mandible, labial palpus and maxillary palpus black. Forewing slightly, rather unevenly brownish, medial and submedial cells mostly uniformly hyaline, infuscate at apex (Figs. 3‒4). Hindwing hyaline, somewhat infuscate at apex (Fig. 4). Hình 3.1 - 3.6: Chalybion tanvinhensis Pham và Ohl, 2019 (female) 3.1. Head, frontal view. 3.2. Mesosoma, dorsal view. 3.3. Right forewing. 3.4. Habitus, dorsal view. 3.5. Habitus, lateral view. 3.6. Metasoma, sternum view. Male (Figs. 7‒10): Body length 14.5-15.5 mm, forewing length 10.5 mm. Structure as in female, but differing as follows: Head (Fig. 7): Mandible without inner subapical tooth. Clypeus without teeth, median and submedian tooth fused into single protruding lobe. Distance between hindocelli 0.62-0.67 times that from hindocellus to inner eye margin and about 0.31-0.33 times that from hindocellus to vertex margin; antenna with placoids on seventh to ninth flagellomeres. Flagellar ratio (length of the first flagellomere divided by that of the second flagellomere) 0.89-0.94; orbital ratio (shortest interocular width across vertex divided by shortest interocular width across clypeus) 0.93-0.97; clypeal ratio (length of clypeus divided by 14 shortest interocular width across clypeus) 0.60-0.67; hypostomal cavity ratio (length of hypostomal cavity divided by width) 0.89-0.96. Mesosoma (Figs 8‒9): Mesopleuron coarsely, densely punctatorugose. Posterior propodeal surface coarsely, densely punctate-reticulate. Mesosoma ratio (length of mesosoma divided by height) 2.18-2.21. Metasoma (Figs. 9‒10): Metasomal sterna IV and V with large patch of micropubescence. Petiole-basitarsal ratio (length of petiole divided by that of hindbasitarsus) 0.58-0.64. Hình 3.7 - 3.10: Chalybion tanvinhensis Pham và Ohl, 2019 (male) 3.7. Head, frontal view. 3.8. Habitus, dorsal view. 3.9. Habitus, lateral view. 3.10. Metasoma, sternum view. Distribution: Vietnam: Hoa Binh, Son La Chalybion tanvinhensis belonged to the C. fabricator species group. This group has 8 species having been described hitherto: C. malignum (Kohl), C. accline (Kohl), C. magnum Hensen, C. sulawesii Ohl, C. lividum Hensen, C. fabricator (Smith) và C. hainanense Terayama and Tano, and C. tanvinhensis Pham and Ohl. 3.1.3. Distribution of solitary wasps of the family Sphecidae at the areas of study Distribution of solitary wasps of the family Sphecidae at 4 habitats was presented in table 3.3. Said results showed 19 species belonging to 7 genera in 4 subfamilies being recorded in the natural forest habitat, 25 species belonging to 7 genera in 4 subfamilies in the artificial forest and orchard habitats, 17 species belonging to 4 genera in 4 subfamilies in the shrub habitat, and 10 species belongting to 4 genera in 4 subfamilies in the resident habitat. Table 3.3. Distribution of solitary wasps of the family Sphecidae in habitats of study No. Species name Habitat Natural forest Artificial forest and orchard Shrub Resident 1 Ammophila clavus + + + + 2 Ammophila globifrontalis - + - - 3 Ammophila laevigata + + + + 4 Chlorion lobatum + + + + 5 Chalybion bengalense + + + + 6 Chalybion dolichothorax + - - - 7 Chalybion gracile - + - - 8 Chalybion japonicum + + + + 9 Chalybion malignum + + + - 10 Chalybion tanvinhensis - + - + 11 Sceliphron deforme + + + - 12.1 Sceliphron javanum petiolare + + - - 12.2 Sceliphron javanum chinensis + + - - 15 13 Sceliphron madraspatanum + + + + 13.1 Sceliphron madraspatanum sutteri - - + + 13.2 Sceliphron madraspatanum andamanicum - + + - 13.3 Sceliphron madraspatanum conspicillatum - - + - 13.4 Sceliphron madraspatanum kohli - + + + 14 Isodontia aurifrons + + - - 15 Isodontia chrysorrhoea + - - - 16 Isodontia elsei - + - - 17 Isodontia nigella + + + - 18 Isodontia sp.1 - - + - 19 Isodontia sp.2 + + - - 20 Isodontia sp.3 + + - - 21 Prionyx viduatus + + + - 22 Sphex argentutus + + + + 23 Sphex diabonicus + + - - 24 Sphex sericeus - + + - 25 Sphex subtruncatus + + + - Tổng 19 25 17 10 Remark: + present, - absent 3.2. Some biological characteristics of S. madraspatanum 3.2.1. nesting construction Materials of nesting construction of S. madraspatanum were mud or coal. The coulour of the nest was based on that of mud, but that of nests known mainly was brown. The nest of the wasp was constructed from 1 to 17 cells. Whereas nests constructed only one cell, no cover was made, but nests with more than 2 cells, a cover was made. Based on nesting sites and nesting substrates, nesting covers thickened variously from 2.5 to 10.5 mm. The outside surface of nests was often rough, but that of some was flat and some nests were with several mud thorns on the nesting surface. On many instances, there were space between pieces of mud put on the nesting surface. Nesing cells were pipe-shaped and round ends. Partitions between nesting cells measured 1.0 – 1.2 mm, average 1.1  0.07 mm. The outside surface of cells was rough, but the into surface was smooth. Nesting cells built adjoining, and partition of one cell became that of the next cell. There was a space between cells, but this space was made fully with mud. Nests of the wasp were usually constructed from 1 – 3 floors, number of cells per floor decreased following floors above. Cell size contained female was often larger than that contained male (table 3.4). Table 3.4. Nesting cell size of S. madraspatanum Length (cm) (n = 32) Width (cm) (n = 32) Female Male Female Male 2.7 – 3.1 2.86  0.12a 2.4 – 2.8 2.63  0.10b 0.7 – 0.9 0.82  0.06c 0.5 – 0.75 0.66  0.07d Remard: In limit of values brough different letters showing confident variousness at probability P  0.05, n was nesting cells tested 16 3.2.2. The developmental time of stages + Egg stage: Egg of S. madraspatanum was white of yellowish, with egg cover smooth and shiny. Eggs were saussage- shaped, round at the two ends, and moderately curved. The developmental time of egg varied 2 - 3 days, average 2.23  0.43 days (table 3.6). + Larva stage: The larvae of S. madraspatanum had three instars. In general, the coulor of larva was yellowish. Dorsal legs were short and coloured yellow. There were clear joints on their body, each bringing one breathing orifice. Skin of larvae smooth. The back of larva was with a ridge longed from the first joint to the anus joint. The developmental time of the 1st instar larva was 1-2 days, average 1.39  0.49 days; the 2st instar larva from 1 - 2 days, average 1.65  0.48 days; the 3st instar larva from 4 - 5 days, average 4.31  0.47 days (table 3.6). + Pupal stage: The pupa of S. madraspatanum was free and was into a cocoon coloured brownish. There were 2 stages: the prepupa and the pupa. The first was cloured yellow and the body had 13 joints ranged from the thorax to abdomen (3 thorax joints and 10 abdomen joints), the largest 6 – 8 joints. The last had morphological body similar to that of adult, excepting clour of pupae newly emerged and thorns distributed the two sides of the body. The developmental time of the prepupa was 5 - 6 days, average 5.58  0.50 days. The developmental time of the pupa was 13 - 16 days, average 14.48  1.14 days (