Summary of the Phd Thesis

MINISTRY OF EDUCATION VIETNAM ACADEMY OF SCIENCE AND TRAINING AND TECHNOLOGY

GRADUATE UNIVERSITY OF SCIENCE AND TECHNOLOGY ************************

PHAM PHONG HUY

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 MADRASPATANUM (FABRICIUS, 1781)

Major: Entomology Code: 9420106

SUMMARY OF THE PHD THESIS

Ha Noi – 2020

The research was completed in Graduate University of Science and Technology – Vietnam Academy of Science and Technology

Supervisor 1: Prof.Dr. Truong Xuan Lam Supervisor 2: PhD. Michael Ohl

Reviewer 1: … Reviewer 2: … Reviewer 3: ….

The thesis will be defended to the doctoral evaluation committee at Graduate University of Science and Technology – Vietnam Academy of Science and Technology at …………………………………... on ……………….……, 2020.

The thesis can be found at: - Library of Academy of Science and Technology - Vietnam National Library

INTRODUCTION 1. Importance of the research Solitary wasps of the family Sphecidae are important in several habitats and most are useful to people’s aim. They are prey specialists and pollinators, but some of them are biomarkers for several ecosystems. Surveys and studies on the composition, biology, ecology and habits of sphecid wasps of this family are necessary and bring scientific and real meanings to highlight their valuableness in ecosystems (Bohart và Menke, 1976; Buck, 2003). Studies on the sphecid wasp fauna of Vietnam are hirtheto limited. These focus only surveys of the composition of the sphecid wasps. 24 species and 3 subspecies of the family Sphecidae are, as to now, recorded for the Vietnamese fauna. The number of sphecid wasps of Vietnam (24) is much less than that listed in the world (789) (Pulawski, 2019). The northwest part includes 6 mountainuos provinces in which there is a variety of climate and topography. Its border line is with Chiness and Lao neighbours and the area is on the eastern side of the Hymalaya range in which there is a high diversity of the fauna and flaura. Hence, the northwest area is expected to contain several new species of the family Sphecidae that have not yet published up to now. The area is, to present, recorded only 6 sphecid wasps (Institution of plant protection, 1969; Mai Quy et al., 1981; Pham et al., 2015). Sceliphron madaspatanum is one of sphecid species of the genus Sceliphron, which has received numerous interests of entomologists, such as: Horne, 1870; Cameron, 1889; Dutt, 1912; William, 1919; Bernard, 1935; Katayama and Ikushima, 1935; Ma, 1936; Hertzog, 1956; Spurway et al., 1964; Iwata, 1964; Myartseva, 1968; Iida, 1969; Begum et al., 1989; Chatenoud et al. 2012; and Gess and Roosenschoon, 2016. Their studies on the wasp pay an attention mainly on records of observation rather than those based on laboratory methods. Therefore, the results are only some nesting habits, larva descriptions, and prey and predator records. Numerous biological, ecological, and behavior charasteristics of the species have not yet been interested in. In north Vietnam, S. madraspatanum is abundant in number and distributed largely. Its nesting sites are easy to find, observe and collect, it is, hence, easy to studies on its biology, ecology and habits (Pham, 2016, Pham et al., 2019). From statements mentioned above. The project is selected and carried out, with title “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 madraspatanum (Fabricius, 1781)”. The project is to produce new data on the composition of sphecid wasps in Vietnam as well as to add new data on biological, ecological, and behavior charasteristics of the mud dauber wasp S. madraspatanum. 2. Aim of the research The project is aimed to yield the composition and distribution of solitary wasps of the family Sphecidae in several provinces of the northwest part of Vietnam, new species to science and new species to Vietnam, and addition of some new biological, ecological, and behavioral characteristics of the mud dauber wasp S. madraspatanum. These results will be scientific base to next studies on the family Sphecida in the future. 3. Research content Content 1: Studying on the composition and distribution of sphecid wasps of the family Sphecidae, describing new species to science and determining new species to several provinces of the northwest part of Vietnam. Content 2: Studying on some biological and ecological characteristics of the mud dauber wasp S. madraspatanum

Content 3: Studying on some behavioural characteristics of the mud dauber wasp S. madraspatanum

CHAPTER 1. OVERVIEW 1.1. Studies on sphecid wasps of the family Sphecidae in the world 1.1.1. Studies on the composition and distribution of sphecid wasps of the family Sphecidae Hitherto, thousands of publications related to sphecid wasps of the family Sphecidae in the world have been issued. All of these have been listed by Pulawski (2019). Following said list, there are 789 species belonging to 19 genera in 5 subfamilies recorded for the family. Table 1.1. Number of species and genera of the family Sphecidae in the world (Pulawski, 2019) No. Scientific name Numbe Geographic distribution r I Subfamily 347 Ammophilinae 1 Genus Ammophila 240 The Oriental region, the Palaearctic region 2 Genus Eremnophila 9 The Nearctic region, The Neotropical region, the Oriental region 3 Genus Eremochares 6 The Palaearctic region 4 Genus 4 The Palaearctic region, The Oriental region, the Hoplammophila Nearctic region, The Neotropical region 5 Genus 21 The Oriental region, the Ethiopian region, the Parapsammophila Australian region 6 Genus Podalonia 67 The Oriental region, the Palaearctic region, the Nearctic region, The Neotropical region, the Ethiopian region II Subfamily 20 Chloriontinae 7 Genus Chlorion 20 The Nearctic region, The Neotropical region, The Oriental region, the Palaearctic region III Subfamily 148 Sceliphrinae 8 Genus Dynatus 3 The Nearctic region 9 Genus Penepodium 22 The Neotropical region 10 Genus Podium 23 The Neotropical region 11 Genus Trigonopsis 16 The Neotropical region 12 Genus Chalybion 49 The Oriental region, the Palaearctic region, the Nearctic region, the Neotropical region, the Ethiopian region 13 Genus Sceliphron 35 The Oriental region, the Palaearctic region, the Nearctic region, the Neotropical region, the Ethiopian region, the Australian region IV Subfamily 273 Sphecinae 14 Genus Chilosphex 2 Tthe Palaearctic region 15 Genus Palmodes 21 The Nearctic region, the Palaearctic region, The Oriental region, the Australian region

16 Genus Prionyx 56 The Oriental region, the Palaearctic region, the Nearctic region, the Neotropical region, the Ethiopian region, the Australian region 17 Genus Isodontia 62 The Oriental region, the Palaearctic region, the Nearctic region, the Australian region. 18 Genus Sphex 132 The Oriental region, the Palaearctic region, the Nearctic region, the Neotropical region, the Australian region, the Ethiopian region. V Subfamily 1 Stangeellinae 19 Genus Stangeella 1 The Neotropical region Total 789 The genus Sceliphron consists of 35 species and 30 subspecies distributed in the whole world. Having 8 subspecies, S. madraspatanum is one of species that contains the most numerous subspecies in the genus Sceliphron. 1.1.2. Studies on biology of the mud dauber wasp S. madraspatanum and other species of the family Sphecidae 1.1.2.1. Studies on nesting construction Publications of Dutt (1912), Myartseva (1968), and Kazenas (2001) showed that a nest of the mud dauber wasp S. madraspatanum has from 2 – 7 cells. Katayama and Ikushima (1935) reported that a nest of S. madraspatanum in Japan has from 5 – 13 cells, including 2 - 3 floors and each bringing 3 - 5 cells. Studies of Camillo (2002) in Braxin on S. fistularium showed the nesting construction of the wasps having 1 - 54 cells, cells sausage-shaped, 20,8 - 29,7 mm long and 7,6 - 11,7 mm in diameter. Following a publication of Kazenas et al. (2004), a nest of S. deforme in Kazahkstan is built from 12 - 15 cells. Gess and Gess (2014) stated that S. spirifex in Africa builds its nest with mud, cells cycle-shaped at the two ends and has the maximum of 12 cells. 1.1.2.2. Studies on the developmental time of immature stages, ovary and sting construction Dutt (1912) is a pioneer studying on the developmental time of immature stages of S. madraspatanum in India. Following his research results, there are 3 developmental stages of the species: the egg stage lasts 1 - 2 days, the larva stage takes 11 - 15 days and the pupa stage lasts 11 - 13 days, the total of time from egg to the adult emerged is 23 - 30 days. Katayama and Ikushima (1935) recorded an adult female of S. madraspatanum laying about 19 eggs during its life time. Ohl and Linde (2003) stated that the number of egg pipes of S. madraspatanum in general, and of sphecid wasps is 6 and there are egg pipes on a egg group. Studies of Basil-Edwardes (1921) in India on S. deforme showed that the developmental time from the egg newly hatched to adult emerged is around 4 weeks. 1.1.3. Studies on the ecology of S. madraspatanum and other species of the family Sphecidae 1.1.3.1. Studies on nesting sites Horne (1870), Cameron (1889), Williams (1919), Katayama and Ikushima (1935), Iwata (1939, 1964), and Begum et al. (1989) stated that nesting sites of S. madraspatanum are in desolated houses or human houses. Freeman (1974) recorded S. assimile in Giamaica nesting under bridges buildings, overhung cliffs, caves and materials of wood or iron… Studies of Callan (1988) resulted that S. formosum in Australia nests on walls of the garage. De Carvalho et al. (2014) reported S. asiaticum in Braxin nesting in a house in the city of Floriano. 1.1.3.2. Studies on prey 3

Reports of Horne (1870), Cameron (1889), Williams (1919), Katayama and Ikushima (1935), Iwata (1939, 1964), and Begum et al. (1989) recorded prey of S. madraspataum are spiders of numerous different genera. Studying on 80 specimens of spider species collected in 5 cells of S. formosum in Australia, Callan (1988) evidenced that they belong to 4 families. Elgar and Jebb (1999), studying on prey deposited in 49 cells of nests of S. laetum in Braxin showed that there are 4 families of Araneidae, Oxyopidae, Salticidae, and Thomosidae, with a totla of about 14 genera. Polidori et al. (2007) judged that prey of S. caementarium and S. spirifex in north Italia belong to 17 families and 49 genera. 1.1.3.3. Studies on predators Katayama and Ikushima (1935) reported that predators of S. madraspatanum are Teirastichus sp. and Chrysis (Tetrachrysis) fuscipesnnis murasaki Uchida in Japan. Studies of Iwata (1939, 1964) proved old nests of S. madraspatanum being utilized by Sceliphron inflexum Sickmann (a synonym of Chalybion japonicum) and Odynerus guttulatus Saussure (a synonym of Stenodyneriellus guttulatus). Krombein and Walkley (1962) showed 3 parasitic wasps of nests of S. spirifex in South Africa: Dolichomutilla minor minor, Chrisis (Pyria) lyncea, and Osprynchotus gigas. Publication of Freeman and Parnell (1973) reported Melittobia chalybii Ashmead causing a dead rate, up to nearly 52.3%, to the developmental stages of S. assimile Dahlbom in Giamaica. Fernández-Triana et al. (2005) recorded nesting parasitic wasps of S. assimile in Cuba including Melittobia sp., Acroricnus cubensis, Chrisis sp., several flies and a species of fungi. 1.1.3.4. Studies on roles of S. madraspatanum Nazarova and Shomirsaidov (1997) and Wu and Zhou (1996) mentioned that S. madraspatanum takes a important role in pollination to plants, specially fruit trees in Russia and China. Studies of Gess and Roosenschoon (2016) showed that S. madraspatanum flies often to flowers of Rumex dentatus to taking nectar. The recent study of Dunford et al. (2014) showed that S. madraspatanum in Afghanistan is one of dangers to soldiers who walk in the forest because it attachs they by sting. 1.1.4. Studies on habits of S. madraspatanum and other species of the family Sphecidae 1.1.4.1. Studies on nesting habits Cameron (1889) and Dutt (1912) observed several adult females of S. madraspatanum aggregating at a small pool near a stream, they using their mandible and a pair of foreleg to roll pieces of mud and then bringing these pieces of mud to its nest. The wasp takes about one day to complete a nest cell. According to a study of Williams (1919) in Philippines, a female of S. madraspatanum does 9 cells in a period of 16 days. Studies of Iwata (1939, 1964) proved that a adult female of S. madraspatanum in Japan builds its nest in sheeted sites. Following research of Hertzog (1956), there are 2 stages of nesting building of S. madraspatanum, the first: collecting mud, the last: building. Study results of Spurway et al. (1964) presented S. madraspatanum in India building its nest in desolated houses. The female brings mud by its mandible and when these pieces of mud are put at nesting site, she uses her a pair of the forleg to drag them out and her mandible to attach them to. Studies of Peckham and Peckham (1898) showed that S. caemenatarium in U.S.A using 36 - 40 pieces of mud to build a nesting cell. 1.1.4.2. Studies on egg habits According to studies of Horne (1870), Williams (1919), Bernard (1935), Ma (1936) and Spurway et al. (1964), after completing building of a nest cell, a female of S. 4

madraspatanum flew away to search its prey. The egg of the species is laid on a first prey brough back to its nest. Callan (1988) stated that S. formosum in Australia, after completing building of a nest cell, a female searchs immidiately prey and brings prey back to its nest, then lay its egg on the first prey placed in the nest cell. Results of a publication of Gess and Gess (2014) produced 3 species S. spirifex, S. quartinae, and S. fossuliferum in South Africa laying egg on the first prey deposited in the nesting cell. 1.2. Studies in Vietnam on the family Sphecidae 1.2.1. Studies on the composition and distribution of sphecid wasps of the family Sphecidae Studies on the sphecid wasp fauna of Vietnam has been limited. To date, there are 27 species and 3 subspecies belonging to 7 genera in 4 subfamilies recorded for the Vietnamese fauna (table 1.3). Table 1.3. Sphecid wasps of the family Sphecidae recorded for Vietnam No. Scientific name Authors and recorded year Distribution Subfamily Ammophilinae I. Genus Ammophila 1 Ammophila clavus Plant Protection Research Bac Thai, Lao Cai, (Fabricius, 1775) Institute (1967-1968), Mai Quy et Nghia Lo, Vinh Phu, al. (1981), Nguyen Lien Phuong Ninh Binh, Dac Lac Thi et al. (2007), Dollfuss (2013), Pham et al. (2015) 2 Ammophila laevigate Plant Protection Research Nghia Lo, Dac Lac Smith, 1856 Institute (1967-1968), Dollfuss (2013), Pham et al. (2015) Phân họ Chloriontinae II. Giống Chlorion 3 Chlorion lobatum Plant Protection Research Ha Tay, Hai Hung, (Fabricius, 1775) Institute (1967-1968), Mai Quy et Lang Son, Ha Nam, al. (1981), Pulawski (2015), Pham Nghe An, Song Be, et al. (2015) Tien Giang, Hai Duong, Nghia Lo Phân họ Sceliphrinae III. Giống Chalybion 4 Chalybion bengalense Mai Quy et al. (1981), Tano và Son La, Vung Tu, Đa (Dahlbom, 1845) Kurokawa (2015) Nang 5 Chalybion dolichothorax Hensen (1988), Pham et al. (2015) Undetermined (Kohl, 1918) 6 Chalybion japonicum Hensen (1988), Barthélémy Ha Noi, Ha Giang (Gribodo, 1880) (2014), Pham et al. (2015) 7 Chalybion gracile Hensen (1988), Pham et al. (2015) Ba Ria –Vung Tau Hensen, 1988 8 Chalybion sumatranum Barthélémy (2014), Pham et al. Undetermined (Kohl, 1884) (2015) IV. Giống Sceliphron 9 Sceliphron clypeatum Pham (2016) Thai Binh Pham, 2016

10 Sceliphron deforme Mai Quy et al. (1981), Hensen Hoa Binh (Smith, 1856) (1987), Pham et al. (2015) 11.1 Sceliphron javanum van der Vecht và van Breugel Ho Chi Minh city, chinense van Breugel, (1968), Párd và Tkalců (1989), Dong Nai 1968 Pham et al. (2015) 11.2 Sceliphron javanum Párd và Tkalců (1989), Pham et al. Dong Nai petiolare Kohl, 1918 (2015) 12 Sceliphron Plant Protection Research Bac Thai, Ha Bac, madraspatanum Institute (1967-1968), Mai Quy et Ha Tay, Hai Hung, (Fabricius, 1781) al. (1981), van der Vecht và van Lang Son, Lao Cai, Breugel (1968), Párd và Tkalců Nam Ha, Nghe An, (1989), Pham et al. (2015) Nghia Lo, Ninh Binh, Thai Binh, Thanh Hoa, Tuyen Quang, Vinh Phu, Song Be, Tien Giang, Ban Me Thuot, Tay Ninh, Phu Quoc, Dong Nai 12.1 Sceliphron van der Vecht và van Breugel Ha Noi, Dong Nai madraspatanum kohli (1968), Párd và Tkalců (1989), Sickmann, 1894 Pham et al. (2015) 13 Sceliphron sp1. Plant Protection Research Nghe An, Dong Nai Institute (1967-1968), (1977- 1978) 14 Sceliphron sp2. Plant Protection Research Vinh Phu, Dong Nai Institute (1967-1968), (1977- 1978) Phân họ Sphecinae V. Giống Prionyx 15 Prionyx viduatus (Christ, Plant Protection Research Ha Tay, Hai Phong, 1791) Institute (1967-1968), Clark Nghe An, Tay Ninh (1987), Danilov (2012), Augul et al. (2015), Pham et al. (2015) VI. Giống Isodontia 16 Isodontia aurifrons Pham et al. (2019) Vinh Phuc (Smith, 1859) 17 Isodontia auripygata Hensen (1991), Pham et al. (2015) Undetermined (Strand, 1914) 18 Isodontia chrysorrhoea Dollfuss (2008), Pham et al. Vinh Phuc (Kohl, 1890) (2015) 19 Isodontia diodon (Kohl, Barthélémy (2014), Pham et al. Undetermined 1890) (2015) 20 Isodontia edax Bohard và Menke (1976), Pham et Undetermined (Bingham, 1897) al. (2015) 21 Isodontia vanlinhi Pham, Pham (2016) Thai Binh 2016

VII. Giống Sphex 22 Sphex argentatus Plant Protection Research Ha Nam, Ninh Binh, Fabricius, 1787 Institute (1967-1968), Nguyen Ba Ria – Vung Tau Lien Phuong Thi et al. (2007), Dollfuss (2008), Barthélémy (2014), Pham et al. (2015) 23 Sphex deplanatus Kohl, Dollfuss (2008), Pham et al. Phan Rang 1895 (2015) 24 Sphex diabolicus Smith, Plant Protection Research Bac Thai, Quang 1858 Institute (1967-1968), Berland Ninh, Lao Cai?, (1928), Dollfuss (2008), Pham et Phan Rang al. (2015) 25 Sphex sericeus Barthélémy (2014), Pham et al. Undetermined (Fabricius, 1793) (2015) 26 Sphex subtruncatus Berland (1928), Pham et al. Undetermined Dahlbom, 1843 (2015) 27 Sphex sp. Plant Protection Research Bac Thai Institute (1967-1968) Notes: ? Undetermined location Table 1.3 shows there are 2 subspecies of S. madraspatanum recorded for the sphecid wasp fauna of Vietnam: S. madraspatanum madraspatanum (Fabricius) và S. madraspatanum kohli, Sickmann. 1.2.2. Studies on biological, ecological, behavioural characteristics of sphecid wasps of the family Sphecidae Up to now, there is only a research related to nesting habits of the wasp Prionyx viduatus (Christ). CHAPTER 2. MATERIALS AND METHODS 2.1 Object of the project Objects of the project were sphecid wasps of the family Sphecidae, S. madraspatanum, prey (spiders) and predators of S. madraspatanum 2.2 Duration, location, and materials of research 2.2.1. Duration of research: 5/2017 - 5/2020 2.2.2. Location of research: Including 4 provinces: Hoa Binh, Son La, Yen Bai và Lao Cai. 2.2.3 Material of research: Including: insect nets, insect boxes, insect pins, microscopes, camera, pens, books, forceps... 2.3. Methods of research 2.3.1. Method of study on the composition and distribution of sphecid wasps of the family Sphecidae 2.3.1.1. Investigation and collection of specimens To collect the maximum of specimens and utilize time of collection trips, we used methods of collection of speciments that have been used by Krombein (1976, 1984) and Pham et al. (2019) including: Insect nets, Malaise traps, Trap nests, nest collection. Rate of collection equals 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

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.. 8

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.

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 10

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

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 12

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

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 Habitat No. Species name Natural Artificial Shrub Resident forest forest and orchard 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 + + - - 14

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 2.4 – 2.8 0.7 – 0.9 0.5 – 0.75 2.86  0.12a 2.63  0.10b 0.82  0.06c 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

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 (table 3.6). + Time before laying egg of adult: The time before laying egg of adult was 5 - 7 days, average 5.87  0.76 days (table 3.7). + Life cycle time: The time of life cycle of S. madraspatanum varied 31 - 41 days, average 35.51  4.27 days. Table 3.6. The developmental time of stages of S. madraspatanum (average temperature 28.1  0.95C and average humidity 80.9%  5.6%) Developental stage Developmental Average time (day) (day) Egg (n = 35) 2 - 3 2.23  0.43 1 instar (n = 46) 1 - 2 1.39  0.49 Larve 2 instar (n = 40) 1 - 2 1.65  0.48 3 instar (n = 35) 4 - 5 4.31  0.47 Prepupa (n = 43) 5 - 6 5.58  0.50 Pupa (n = 52) 13 - 16 14.48  1.14 Time before laying egg of adult (n = 23) 5 - 7 5.87  0.76 Life cycle 31 - 41 35.51  4.27 Remark: n was the number of specimens tested 3.2.3. Living time of adult Under laboratory condition, average temperature 28.6  1.02C and average humidity 81.2  3.45% (table 3.7), adults reared by 50%diluted bee’s honey, the living time of male varied 11 - 27 days, average 18.5  3.61 day and that of female 31 - 71 days, average 52.0  11.1 days. Table 3.7. Living time of adult of S. madraspatanum reared by 50%-diluted bee’s honey No. Living time (days) Average Average Male Female temperature humidity (%) (C) 1 11 - 25 31 - 69 29.8 78.3 (n = 30) 18.5  4.49a 51.2  9.80b 16

2 13 - 25 31 - 71 28.2 81.1 (n = 30) 18.2  3.03a 51.6  11.6b 3 13 - 27 31 - 70 27.8 84.2 (n = 30) 18.7  3.32a 53.2  11.8b Trung 11 - 27 31 - 71 27.8 – 29.8 78.3 – 84.2 bình 18.5  3.61 52.0  11.1 28.6  1.02 81.2  3.45 Remard: In limit of values brough different letters showing confident variousness at probability P  0,05, n was specimens tested 3.2.4. Living rate of immature stages With tests repeated the 3 times showed that rate of hatched egg in first time was 93.5%, the second 98.4% and the last 93.6 %. Therefore, the rate of hatched egg varied 93.5 – 98.4%, average 95.2% (table 3.8). The living rate of larva between the 3 times of test was 80.7%, 80.4% and 87.3%, respectively. Hence, The living rate of larva ranged 80.4 – 87.3%, average 82.8%. The living rate of pupa between the 3 times of test was 86.3, 90.3%, and 77.9%, respectively. Hence, The living rate of pupa varied 77.9 – 90.3%, average 84.8%. Table 3.8. Living rate of immature stages of S. madraspatanum No. Living rate of stages (%) Temperature Humidity Egg Larva Pupa (C) (%) 1 93.5 80.7 86.3 29.1 82.3 (n = 31) (n = 83) (n = 102) 2 98.4 80.4 90.3 28.4 83.3 (n = 62) (n = 97) (n = 113) 3 93.6 87.3 77.9 26.9 77.1 (n = 47) (n = 63) (n = 77) Average 95.2 82.8 84.8 28.1  0.95 80.9  5.6 Remark: n was number of specimens tested 3.2.5. Sex rate We collected nests of S. madraspatanum in the field and then observed them under the laboratory condition (average temperature 28.1C and average humidity 80.9%) showing in 49 nests observed, 3 of which had only males, 5 had only females and the remaining had both sexes (table 3.9). Table 3.9. Emerging order of both sexes on nests of S. madraspatanum Nests observed Number of adults Sex Sex rate (♀/♂) 1 and 9 2 ♀,♂ 1:1 2, 19 and 20 6 ♀,♂,♀,♀,♂,♀ 1:0.5 3, 4, 6 and 11 5 ♂,♀,♀,♂,♂ 1:1.5 3, 4, 6 and 11 4 ♀,♂,♂,♀ 1:1 5 and 16 4 ♀,♂,♀,♂ 1:1 7 and 8 4 ♀,♀,♀,♂ 1:0.33 10 6 ♂,♂,♀,♂,♀,♀ 1:1 12 and 15 5 ♀,♂,♂,♂,♂ 1:4 13 and 18 10 ♀,♂,♀,♂,♀,♂,♂,♀,♂,♀ 1:1 14 8 ♂,♀,♀,♀,♀,♂,♀,♂ 1:0.60 17 and 21 11 ♂,♂,♂,♀,♀,♀,♀,♀,♀,♂,♀ 1: 0.57 Average 1:1.14 17

Remark: ♀: female, ♂: male 45 40 60 Cái 35 50 30 40 Đực 25 30 20 20 10 15 (%)Rate Number of Number 0 10 Tỉ lệ ngoài Tỉ lệ trên Tỉ lệ trong 5 tự nhiên 21 tổ các đợt 0 nhân nuôi Tổ chỉ có giới Tổ chỉ có giới Tổ có cả hai tính đực tính cái giới tính

Figure 3.26. Rate on nests of S. Figure 3.27. Sex rate of S. madraspatanum madraspatanum Obervations on 41 nests with both sexes, 21 had a sexes rate 1: 1,14 (♀/♂) (table 3.9, figure 3.27). The sex rate based on results tested in the laboratory was (♀/♂) = 1:1,44 and that in the field (♀/♂) = 1: 1,11 (figure 3.27). 3.3. Some ecological characteristics of S. madraspatanum 3.3.1. Nesting site Nesting sites of S. madraspatanum were firmed substances because these sites were protected from the sun and rain. These were mainly at buildings, prominent cliffs, desolated houses. Surveys at 313 nesting sites of the wasp showed there were 10 main nesting sites, including: 1- walls of houses, 2- windows, 3- Roofs, 4- The lower surface of floors, 5- Floors, 6- fences, 7- housewares, 8- under bridges, 9- power poles, 10- prominent cliffs. 3.3.2. Overwintering time Overwitering nests of S. madraspatanum built from the end of September to the beginning of October and prepupae appeared at the end of October. These prepupae took an overwintering time until April of the following year (table 3.11). Table 3.11. Overwintering time of S. madraspatanum Study Overwintering time (day) Average (day) Average Average year Shortest Longest temperature humidity (C) (%) 2016 167 176 171.7  3.2 19.3 81.5 (n = 18) 2017 171 184 177.7  3.9 20.5 82 (n = 23) 2018 161 181 170.5  7.1 20.4 79 (n = 41) 2019 166 178 171.4  4.0 18.6 83 (n = 33) Average 166.3 179.7 172.8  4.5 19.7 81.4 Remark: n was number of specimens tested 3.3.3. Occuring time of adults in year Occurrence of adults of S. madraspatanum in year observed from the beginning of April to the end of November during 3 years 2017 – 2019 with 32- 46 nests/year resulted in table 3.12. Sceliphron madraspatanum occurred early on 23 April and dissapearence as late

as on 6 November. The total of occurring time in year of the adult wasp was 192 - 197 days, average 194 ± 9,12 days. Table 3.12. Occurring time of the adult wasp S. madraspatanum in year Year of test Early occurring time Ending time of Total time of in year (day/month) occurrence occurrence in year (day/month) (day) 2017 (n = 46) 23/4 6/11 197 2018 (n = 42) 25/4 2/11 192 2019 (n = 32) 26/4 5/11 193 Average 194 ± 9.12 remark: n was number of nests tested 3.3.4. Effects of food to the living time of adult Different foods affected to the different living time of adult of S. madraspatanum. Following results showed at table 3.15, with food of 50%-diluted bee’s honey and fruit sugar cane, the living time of the wasp was the same. Table 3.15. Effects of food to the living time of adult wasps of S. madraspatanum Sex tested Thời gian sống của trưởng thành No food Fresh water 50%-diluted Fruit sugar bee’s honey cane Male (n = 30) 1 – 2 3 - 5 11 – 25 11 – 25 1.5  0.51a 3.90  0.80b 18.5  4.49c 18.3  3.50c Female (n = 1 - 3 3 - 5 31 - 69 31 - 66 30) 2.03  0.81d 4.27  0.74b 51.2  9.80e 50.1  10.10e Remark: In limit of values brough different letters showing confident variousness at probability P  0,05, 3.3.5. Prey Prey of S. madraspatanum were spider of belonged to 4 families into 8 genera. In which, 5 genera were of the family Araneidae, one genus of the Oxyopidae, one of the fafmily Tetragnathidae, and one of the falimy Therididae (table 3.14). Compared to results having been studied before, the present study recorded 7 genera belonging to 3 familes as new prey to S. madraspatanum. Table 3.14. The composition of prey of S. madraspatanum No. Scientific name Finding rate on nest (%) I. Họ Araneidae 1 Genus Araneus Clerck, 1757 80 2 Genus Cyclosa Menge, 1866 20 3 Genus Cyrtophora Simon, 1864 30 4 Genus Leucauge, White, 1841 30 5 Genus Zygeilla Cambridge, 1902 50 II. Họ Oxyopidae 6 Genus Oxyopes Latreille, 1804 10 III. Họ Tetragnathidae 7 Genus Tetragnatha Latreille, 1804 30 IV. Ho Therididae 8 Genus Therodion Walckenaer, 1805 40

3.3.6. Predators Predators of S. madraspatanum showed in table 3.17. Of which, 9 families, 12 genera, and 13 species were predators of S. madraspatanum. Pison punctifron of the family Crabronidae, Antepipona excelsa keralensis, Euodynerus trilobus of the family Vespidae, Orphnaeus brevilabiatus of the family Oryidae, and Hemidaciylus frenatus of the family Gekkonidae were recorded as new predators of S. madraspatanum. The genus Melittobia (the parasite Melittobia clavicornis (Cameron, 1908)) was recorded for the first time to the parasitic fauna of Vietnam. Table 3.17. Predators of S. madraspatanum No. Scientific name Parasitic stage I Family Sphecidae 1 Chalybion japonicum (Gribodo, 1880) Egg, larva 2 Chalybion bengalense (Dahlbom, 1845) Undetermined II Family Crabronidae 3 Pison punctifron Shuckard, 1838 Egg, larva III Family Apidae 4 Amegilla zonata (Linnaeus, 1758) Egg, larva, pupa IV Family Ichneumonidae (họ ong cự) 5 Enicospilus flavocephalus (Kirby, 1990) Larva 6 Euchalinus sp. larva V Family Vespidae 7 Antepipona excelsa keralensis Lamber, 2004 Egg, larva, pupa 8 Euodynerus trilobus (Fabricius, 1787) Egg, larva, pupa VI Family Eulophidae 9 Melittobia clavicornis (Cameron, 1908) Pupa VII Family Bombyliidae 10 Anthrax sp. Larva VII Family Chrisiidae 11 Chrisis sp. Egg, larva VIII Family Oryidae 12 Orphnaeus brevilabiatus Newport, 1845 Undetermined IX Family Gekkonidae 13 Hemidaciylus frenatus Schlegel, 1836 Adult 3.4. Some habits of S. madraspatanum 3.4.1. Nesting habits Eight females of S. madraspatanum found nesting at Tan Thanh, Luong Son, Hoa Binh in 2017 and 18 females saw nesting at Hong Minh, Hung Ha, Thai Binh in 2017 and 2018. Females searched mud at pools, and used their pair of for legs to make pieces of mud. These pieces of mud were spherical, 2,0 - 2,5 mm in diameter. Females used their mouth to bring them to nesting sites. Pieces of mud were attched to substrates closely. Females made high buzzes during ther nesting time. At the base of nests, females used 5 - 6 pieces of mud, and when building partitions of cells, females used their mouth and one pair of forelegs to long mud. When size of cells was full, females made nesting entrances narrowed and smooth. Prey were closely put into nest cells. Completing time of one cell was 4522 seconds as showed in table 3.17 and equaled time taken to complete one cell + time taken to provisioning prey + time taken to laying egg.

On 11 cases, Provisioning time was from 4 hours and 21 minutes to 7 hours and 32 minutes, average 6 hours and 4 minutes (21.840 seconds). Egg-laid time was known, but it was about 10 seconds. Therefore, Nest completing time = 4522 + 21.840 + 10 = 26.372 seconds (abour 7 hours and 32 minutes). Females of S. madraspatanum usually made aggregarious nests at sites in which adults had emerged. Many nests of the wasp aggregated and took nest groups. Table 3.17. Completing time of one cell of S. madraspatanum Time taking mud + Time Time attaching mud Time completing No. bringing mud (second) (second) one cell (second) 1 (5/2017) 38 – 82 37 – 79 4447 (n = 38) 60.37  10.42 56.66  12.61 2 (7/2017) 32 – 73 39 – 80 4597 (n= 39) 55.85  10.10 62.02  10.07 3 (8/2018) 35 – 75 48 – 80 4522 (n = 36) 59.05  9.85 66.58  8.12 Average 32 – 82 37 – 80 4522 58.42  10.12 61.75  10.27 Remark: n was times tested 3.4.2. Ovipositioning habits Females of S. madraspatanum laid only one egg per one nesting cell. The egg attched on abdomen of prey, usually on the left side (n = 37). One end of egg was glued to abdomen of prey, one end extending after. The egg was glued on the first, the second or the third prey brough back to the nest (n = 48), on some cases it was on the fifth (n = 3). Laying egg was taken outside or inside nest cells remained unknow. Prey attached egg were not the largest ones provisioned into the cell. 3.4.3. Feeding habits The first instar larva of S. madraspatanum only took solution of prey (n = 46). The second instar larva began to feed on prey (n = 40). Firstly, it fed on the remaining of the first prey, then next prey into cell. The first instar consumed only one prey, the third instar consuming most of prey put into cell. The last instar had the biggest body when it consumed all prey. After stopping feed, the last larva made its cocoon with silk from its mouth. It made continously its cocoon and completed it about 1 day. After completing the cocoon, the larva elimilated feces. The feces was black and lied at the bottom of the cocoon. It was hard then. Study on 109 nesting cells of S. madraspatanum showed that the number pf prey provisioned in male cells was less than that of female cells (table 3.18). Table 3.18. Number of prey provisioned in nesting cell of S. madraspatanum Year of study Number of prey provisioned in one nesting cell (individual) Male cell Female cell 2017 12 - 23 15 - 29 n = 32 17.40  3.84 19.81  3.71 2018 9 - 24 15 - 27 n = 33 16.15  4.37 19.39  3.16 2019 10 - 22 17 - 31 n = 44 15.45  3.77 22.54  4.34 Average 9 - 24 15 - 31 16.33  3.99 20.58  37.36 Remark: n was number of nesting cells tested 21

3.4.4. Emerging habits of adult wasp Before emerged, adult wasps made high buzze, then used a pair of forlegs to break the cocoon (n = 32). The cocoon was splited at the center part. The adult exited from this slit and made the cocoon becoming two parts. The adult bited mud cell to make an emerging hole on the surface of nest. The adult went out the cell and cleaned its antennae and forlegs, midlegs and hindlegs, then it flew away to search food. 3.4.5. Mating habits Males usually had the body size smaller than that of females. The male much often flew around the nesting site, when it found female, immidiately grasped the female from the back. It used its mandible to bite the pronotum, placing his forelegs on both sides of her mesosoma just anterior to her hindlegs, and his midlegs and hindlegs straddling her abdomen. The male tapped his antennae on the top of her head while probing the apex of her abdomen with his genital claspers, then bent his abdomen and attempted to clasp the female’s genitalia from above. The male mated the female continously. During the copulation they often buzzed loudly. We did not know whether or not the buzz that was from the male or the female made. The copulation time varied between 20 second to 8 minutes. After completing the couple, immidiately both the male and female flew away.

CONCLUSIONS AND RECOMMENDATIONS 1. CONCLUSIONS 1. At several provinces of the northwest part of Vietnam, 25 species and 6 subspecies belonging to 7 genera into 4 subfamilies of the family Sphecidae were recorded. Of which, one species had been described to science Chalybion tanvinhensis Pham và Ohl, 2019; 4 species and 3 subspecies were recorded for the first time for the sphecid wasp fauna of Vietnam, and 9 species and 6 subspecies were newly recorded in northwest Vietnam. The genus Melittobia was new recorded for the parasitic wasp fauna of Vietnam. 2. In the conditions of the average temperature of 28.1  0.95C and average humidity of 80.9  5.6%, S. madraspatanum had four developmental stages. Of which, the egg stage had the average developmental time of 2.23  0.43 days, that of the larva stage 7.35  1.14 days, that of the pupa stage (including prepupa and pupa stages) 20.06  1.64 ngày, that of adult before laying egg 5.87  0.76 days. The time of life cycle was 31 - 41 days, average 35.51  4.27 days. Egg-laid rate was 95.2%, the living rate of larva 82.8% and that of pupa 84.8%. The sex rate in the laboratory condition was 1:1.44 (♀/♂) and that in the field was 1:1.12 (♀/♂). 3. Materials used to build nests of S. madraspatanum were mud or coal. The nest of the wasp constructed 1-17 cells neighborly or accumulatively. There were 1-3 nesting floors. 10 nesting sites of the species were recorded, mainly on wall of desolated houses. Sceliphron madraspatanum overwinterred as prepupae with the average time of 172.8  4.5 days (Beginning from the end of October to the beginning of following April). A total of time of occurrence of S. madraspatanum in year was 194 days. Under the laboratory condition, average temperature 27.3  1.27C and average 78.5  7.3%, with food of 50% diluted bee’s honey, the average life time of males 18.5  4.49 days and that of females 51.2  9.80 days. Prey of S. madraspatanum were spiders (8 genera into 4 families), of which, two genera Araneus and Zygeilla were used commonly. 13 species belonging to 12 genera into 9 families were predators of S. madraspatanum, of which, 7 families belonged to the insect class, one family of the chilopoda class, and one family of the mammalia class.

4. Females of S. madraspatanum usually chose nesting sites with an elevation of from 1 to 4 meter. Females used its mouth and for legs to made and bring pieces of mud from mud resources to nesting sites. During building, the wasp always buzzed loudly. 36 - 39 pieces of mud were used to build one nesting cell with the total of building it about 7 hours and 32 minutes. The insect laid only an egg on a first, a second or a third prey brough back to its nest. The 1st instar took solution of prey, the 2st to 3 rd instars consuming prey by chewing. The average number of prey provisioned per one male cell was 16,33  3,99 prey and that provisioned per female cell 20,58  37,36 prey. After emerged, adults buzzed loudly and used a pair of for legs to break cocoons, then used the the mandibles to bite mud cells, and then went out. Females usually cleaned its body with the mouth and the for legs and flew to flowers for food. When mating, the male tapped his antennae on the top of her head while probing the apex of her abdomen with his genital claspers, then bent his abdomen and attempted to clasp the female’s genitalia from above. During the copulation they often buzzed loudly. The copulation time varied between 20 second to 8 minutes.

2. RECOMMENDATIONS It really needs to take added studies on reproductive habits of male of S. madraspatanum such as territorial, mating competition habits and so on as well as methods of attack of predators on nests of S. madraspatanum in the field and the relationship between prey of the wasp and these predators.

NEW CONTRIBUTIONS OF THE THESIS

1- Provided a first list of solitary wasps of the family Sphecidae (Hymenoptera: Apoidea) for several provinces of the northwest part of Vietnam, of which one new species having been published and 4 new species and 3 new subspecies having been recorded for the sphecid wasp fauna of Vietnam. 2- Presented several new data on the biology, ecology and habits of the mud dauber wasp S. madraspatanum. 3- Recorded one new genus Melittobia (Hymenoptera: Eulophidae) for the parasitic wasp fauna of Vietnam.

PUBLICATIONS RELATED TO THE PHD. THESIS

A. PAPERS BELONGING TO THE ISI LIST 1. Phong Huy Pham, 2019. Notes on nest and gregarious behavior of a blue nest-renting wasp, Chalybion japonicum (Gribodo), with comments on commonly used name for Chalybion species (Hymenoptera: Sphecidae). Entomologica Americana, 124 (1-4): 35- 38. http://doi.org/10.1664/1947-5136-124.1.35 2. Phong Huy Pham, 2019. The nesting habits of Chalybion bengalense (Dahlbom) (Hymenoptera: Sphecidae). Oriental Insects, 53(3): 1-12. https://doi.org/10.1080/00305316.2019.1624220 3. Phong Huy Pham, Michael Ohl, Lam Xuan Truong, 2019. The genus Chalybion Dahlbom, 1843 (Hymenoptera: Sphecidae) from Northwest Vietnam, with description of a new species. Zootaxa, 4712(2): 202-210. http://doi.org/10.11646/zootaxa.4712.2.2 B. PAPERS BELONGING TO THE SCOPUS LIST 4. Phong Huy Pham, Hoa Thi Dang, 2017. New record of the mud dauber wasp Chalybion malignum (Kohl, 1906) (Hymenoptera: Sphecidae) from Vietnam with the first information on its nesting biology. Punjab University Journal of Zoology, 32(1): 155-158. 5. Phong Huy Pham, Lam Xuan Truong, Tu Anh Thi Nguyen, 2019. First report of the genus Melittobia Westwood, 1848 (Hymenoptera: Eulophidae) from Vietnam, with notes on the biology of M. clavicornis (Cameron, 1908). Russian Entomological Journal, 28(4): 421–424. http://doi.org/10.15298/rusentj.28.4.10 C. ANOTHER PAPER 6. Phong Huy Pham, 2018. Chalybion bengalense (Dahlbom, 1845) (Hymenoptera: Sphecidae) first reported in North Vietnam. Linzer Biologische Beiträge, 50(1): 681-685.