Spermatogenic cyst and organ culture in <Emphasis Type="Italic">Drosophila pseudoobscura</Emphasis>

Spermatogenesis is a complex series of processes that involves (1) the maintenance of a renewable pool of diploid stem cells within a niche, (2) the mitotic expansion of a subpopulation of stem cells committed to the spermatogenic pathway, and (3) the differentiation of diploid cells into highly specialized, haploid spermatozoa through meiotic and post-meiotic cellular transformations. Drosophila melanogaster is a desirable model for studying spermatogenesis, as similarities exist between mammalian and fly spermatogenesis. Like mammals, flies maintain a spermatogenic stem cell niche; the steps involved in mammalian spermatogenesis are mimicked in flies, with the main difference being that fly sperm develop within cysts rather than an epithelial cell layer. Here, we report a reliable robust system for culturing whole testes and individual spermatogenic cysts obtained from mid- to late-pupal stages of Drosophila pseudoobscura. D. pseudoobscura testes can be easily distinguished in later pupal stages because of their intense red pigmentation and are easily handled because of their simple ellipsoidal morphology. Cultured cysts are comparable in length to cysts obtained from adult flies, and motility is consistently achieved in vitro. This system not only offers a method for dissecting the mechanisms involved in meiotic and post-meiotic cellular transformations, but also can be used for the study of signaling during spermatogenesis.     

Beziehungen zwischen Vermehrungswachstum und Differenzierung von männlichen Keimzellen von Drosophila melanogaster

Summary A count-chamber like methodes enables us to estimate the number of cysts corresponding to each developmental stage of the spermatogenesis.The multiplication of the germ cells depends upon the physiological conditions of the host: it is stopped in isolated larval abdomina, and is low in larvae of a crowded culture where pupation is delayed. In implants of 48 h old testis in. virgin adult females the multiplication rate is lower than in males and in mated females; it is as high as in situ in castrated females or in pupal hosts.The time required for the oldest cysts with spermatocytes to enter the meiotic divisions is with one exception for all these experiments the same as in situ; only in pupal hosts it will be accelerated. The subsequent spermatid differentiation in sperm also takes the same time as in situ. Thereby the premeiotic growth and postmeiotic differentiation is as efficient in all tested milieus as in situ, even when the hosts have not attained or already bypassed the metamorphosis. This autonomy is also valid for germ cells under insufficient multiplying conditions, provided that the spermatocytes have reached a certain stage in the premeiotic growth.When the first cysts reach the postmeiotic stages the further multiplication of cysts is stopped. This regulative mechanism is attributed to a specific inhibition operated by the spermatids.The autonomy of the spermatocyte differentiation and the unified behaviour of all the cells within a cyst are discussed.

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Herrn Prof. Dr. E.Hadorn bin ich zu Dank verpflichtet für die kritische Diskussion dieser Arbeit, Herrn Prof. Dr. H.Burla für die Unterstützung in statistischen Prägen, Herrn Dr. R.Nöthiger und Herrn W.Gehring für die Hilfe bei der Abfassung des deutschen Textes.     

Preference induction and the benefits of floral resources for a facultative florivore

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A New Species of Kudoa (Myxosporidea: Chloromyxidae) from the Spot,Leiostomus xanthurus Lacépède,in Clear Lake,Texas*

SYNOPSIS. Kudoa branchiata sp. n. (Myxosporidea: Chloromyxidae) is described from the gills of the marine sciaenid fish, Leiostomus xanthurus Lacépède, from Clear Lake, Texas. Twelve of the 429 hosts examined from 10 September 1970 to 28 April 1971, were infected. Eight of the 12 infected fish were collected in February and March 1971—a period during which only 13% of the total host sample was taken. The mean total length of infected hosts was 149 mm, with a range of 112–185 mm. The mean number of myxosporidean cysts per infected host was 3.5, with a range of 1–11.     

Host location by the fruit fly parasitoid Diachasmimorpha krausii: role of fruit fly species,life stage and host plant

• 1 Diachasmimorpha krausii is a braconid parasitoid of larval tephritid fruit flies, which feed cryptically within host fruit. At the ovipositor probing stage, the wasp cannot discriminate between hosts that are physiologically suitable or unsuitable for offspring development and must use other cues to locate suitable hosts.
• 2 To identify the cues used by the parasitoid to find suitable hosts, we offered, to free flying wasps, different combinations of three fruit fly species (Bactrocera tryoni, Bactrocera cacuminata, Bactrocera cucumis), different life stages of those flies (adults and larvae) and different host plants (Solanum lycopersicon, Solanum mauritianum, Cucurbita pepo). In the laboratory, the wasp will readily oviposit into larvae of all three flies but successfully develops only in B. tryoni. Bactrocera tryoni commonly infests S. lycopersicon (tomato), rarely S. mauritianum (wild tobacco) but never C. pepo (zucchini). The latter two plant species are common hosts for B. cacuminata and B. cucumis, respectively.
• 3 The parasitoid showed little or no response to uninfested plants of any of the test species. The presence of adult B. tryoni, however, increased parasitoid residency time on uninfested tomato.
• 4 When the three fruit types were all infested with larvae, parasitoid response was strongest to tomato, regardless of whether the larvae were physiologically suitable or unsuitable for offspring development. By contrast, zucchini was rarely visited by the wasp, even when infested with B. tryoni larvae.
• 5 Wild tobacco was infrequently visited when infested with B. cacuminata larvae but was more frequently visited, with greater parasitoid residency time and probing, when adult flies (either B. cacuminata or B. tryoni) were also present.
• 6 We conclude that herbivore‐induced, nonspecific host fruit wound volatiles were the major cue used by foraging D. krausii. Although positive orientation to infested host plants is well known from previous studies on opiine braconids, the failure of the wasp to orientate to some plants even when infested with physiologically suitable larvae, and the secondary role played by adult fruit flies in wasp host searching, are newly‐identified mechanisms that may aid parasitoid host location in environments where both physiologically suitable and unsuitable hosts occur.

     

The spatial patterns of parasitism of eumenid wasps,Anterhynchium flavomarginatum andOrancistrocerus drewseni by the miltogrammine flyAmobia distorta

1. Spatial patterns of parasitism of eumenid wasps Anterhynchium flavomarginatum and Orancistrocerus drewseni by the miltogrammine fly Amobia distorta were studied in Kyoto, Japan during 1980–1984.
2. In generations of low (<5%) and medium (5–20%) parasitism, percent parasitism per shed (the habitat of the hosts) increased as a function of host density. Conversely, in generations of high (>20%) parasitism, percent parasitism was rather constant over different host densities.
3. The spatial distributions of adult miltogrammine flies among sheds were censused in generations of low and medium parasitism. The frequency of observations of adult miltogrammine flies was higher at sheds of higher host density (aggregative behavioral response), but on the other hand, the adult miltogrammine flies distributed in an underdispersed (or regular) manner in relation to other conspecifics.
4. The spatially density independent relationship between host density and percent parasitism in generations of high parasitism was explained in relation to parasitoid dispersal from patches of high parasitoid density.

     

Spotted wing drosophila (Drosophila suzukii) utilization and dispersal from the wild host Asian bush honeysuckle (Lonicera spp.)

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Effect of the <Emphasis Type="Italic">Drosophila</Emphasis> endosymbiont <Emphasis Type="Italic">Spiroplasma</Emphasis> on parasitoid wasp development and on the reproductive fitness of wasp-attacked fly survivors

In a previous study, we showed that Spiroplasma, a maternally transmitted endosymbiotic bacterium of Drosophila hydei, enhances larval to adult survival of its host when exposed to oviposition attack by the parasitoid wasp Leptopilina heterotoma. The mechanism by which Spiroplasma enhances host survival has not been elucidated. To better understand this mechanism, we compared the growth of wasp larvae in Spiroplasma-infected and uninfected hosts. Our results indicate that wasp embryos in Spiroplasma-infected hosts hatch and grow normally for ~2 days, after which their growth is severely impaired, compared to wasps developing in uninfected hosts. Thus, despite their reduced ability to complete development in Spiroplasma-infected hosts, developing wasps may exert fitness costs on their hosts that are manifested after host emergence. The severity of these costs will influence the degree to which this protective mechanism contributes to the long-term persistence of Spiroplasma in D. hydei. We therefore examined survival to 10-day-old adult stage and fecundity of Spiroplasma-infected flies surviving a wasp treatment. Our results suggest detrimental effects of wasp attack on longevity of Spiroplasma-infected adult flies. However, compared to Spiroplasma-free flies exposed to wasps, Spiroplasma-infected flies exposed to wasps have ~5 times greater survival from larva to 10 day-adult. The relative fecundity of wasp-attacked Spiroplasma-infected females was ~71% that of un-attacked Spiroplasma-free females. Our combined survival and female fecundity results suggest that under high wasp parasitism, the reproductive fitness of Spiroplasma-infected flies may be ~3.5 times greater than that of uninfected females, so it is potentially relevant to the persistence of Spiroplasma in natural populations of D. hydei. Interestingly, Spiroplasma-infected males surviving a wasp attack were effectively sterile during the 3-day period examined. This observation is consistent with the expectation that, as a maternally transmitted symbiont, there is little selective pressure on Spiroplasma to enhance the reproductive fitness of its male hosts.     

Infection with Haemoproteus iwa affects vector movement in a hippoboscid fly—frigatebird system

Haemosporidian parasites, which require both a vertebrate and invertebrate host, are most commonly studied in the life stages occurring in the vertebrate. However, aspects of the vector's behaviour and biology can have profound effects on parasite dynamics. We explored the effects of a haemosporidian parasite, Haemoproteus iwa, on a hippoboscid fly vector, Olfersia spinifera. Olfersia spinifera is an obligate ectoparasite of the great frigatebird, Fregata minor, living among bird feathers for all of its adult life. This study examined the movements of O. spinifera between great frigatebird hosts. Movement, or host switching, was inferred by identifying host (frigatebird) microsatellite genotypes from fly bloodmeals that did not match the host from which the fly was collected. Such host switches were analysed using a logistic regression model, and the best‐fit model included the H. iwa infection status of the fly and the bird host sex. Uninfected flies were more likely to have a bird genotype in their bloodmeal that was different from their current host's genotype (i.e. to have switched hosts) than infected flies. Flies collected from female birds were more likely to have switched hosts than those collected on males. Reduced movement of infected flies suggests that there may be a cost of parasitism for the fly. The effect of host sex is probably driven by differences in the sex ratio of bird hosts available to moving flies.     

Mutual interference in the mass-reared fruit fly parasitoid,Diachasmimorpha longicaudata (Hymenoptera: Braconidae)

Diachasmimorpha longicaudata is a solitary endoparasitoid produced in Mexico for the biological control of Anastrepha fruit flies (Diptera: Tephritidae). We studied the effect of mutual interference among conspecific foraging females to better understand the parasitoid-host dynamics established in the mass-rearing system of this species. We used a constant host availability of 60 third instar larvae of Anastrepha ludens (per oviposition unit type Petri dish) that were individually exposed for 3 h to 1, 2, 4, 8, 10, 14, and 20 D. longicaudata females, seven days old without oviposition experience. The following parameters were evaluated: total number of attacked hosts (i.e., larvae with at least one oviposition scar), number of attacked hosts per female, adult emergence percentage, and female proportion. We also performed a second assay at mass-rearing level, contrasting the current proportion used in mass-rearing (~?two host larvae per female) with the alternative treatments (5 and 7.5 host larvae per female). Results showed that the density of females foraging on a patch exhibits an inverse relationship to the number of hosts attacked by one female. The highest values for adult emergence (73.4%), patch exploitation (94.6%), and female proportion (0.86) were obtained in the treatments with 7.5, 6, and 3 host larvae per female, respectively. Under mass-rearing conditions, we found that a lower density of females per cage (~?five larvae per female) reduced superparasitism levels without affecting adult emergence and the proportion of females. This suggests that the mass-rearing efficiency of D. longicaudata could be improved by reducing the density of foraging females.

     

Survival and reproductive rate of mites in relation to resistance of their barn swallow hosts

Parasite resistance may act via a number of different mechanisms that regulate or control the survival and the reproductive rate of parasites. Observations and experiments were used to test for effects of host resistance on parasite survival and rate of reproduction. Natural levels of infestation of barn swallow Hirundo rustica nests by the tropical fowl mite Ornithonyssus bursa were positively related to brood size, inversely related to the length of the outermost tail feathers of male nest owners (a secondary sexual character) and affected by time of reproduction by the host. A mite inoculation experiment, in which 50 adult mites were introduced into nests during the laying period of the host, was used to test for differential survival and reproduction of mites as a function of host resistance. The relationship between survival and reproduction of parasites, male tail length and host resistance was investigated. There was a negative relationship between mite numbers per nest after fledging of nestlings and male tail length. This relationship was mainly caused by a reduction in the number of mites in the first and second nymph stage with increasing tail length of male hosts, implying a reduction in rate of reproduction of mites. The proportion of mites that had recently fed was inversely related to tail length of male hosts. The proportion of nymph stages was positively related to the proportion of mites that had recently had a blood meal. Parasite resistance of barn swallows to the tropical fowl mite thus appeared to act through increased mortality rate of adult and nymph stages of mites, and through reduced reproductive rates of mites on resistant hosts. This is the first study demonstating a direct relationship between fitness components of a parasite and the expression of a secondary sexual character of a host. Received: 11 January 2000 / Accepted: 22 March 2000     

Sertoli cell proliferation in the adult testis--evidence from two fish species belonging to different orders

Germ cell survival and development critically depend on the cells' contact with Sertoli cells in the vertebrate testis. Fish and amphibians are different from mammals in that they show a cystic type of spermatogenesis in which a single germ cell clone is enclosed by and accompanied through the different stages of spermatogenesis by an accompanying group of Sertoli cells. We show that in maturing and adult testes from African catfish and Nile tilapia, Sertoli cell proliferation occurs primarily during spermatogonial proliferation, allowing the cyst-forming Sertoli cells to provide the increasing space required by the growing germ cell clone. In this regard, coincident with a dramatic increase in cyst volume and number of germ cells per cyst, in Nile tilapia, the number of Sertoli cells per cyst was strikingly increased from primary spermatogonia to spermatocyte cysts. In both African catfish and Nile tilapia, Sertoli cell proliferation is strongly reduced when germ cells have proceeded into meiosis, and stops in postmeiotic cysts. We conclude that Sertoli cell proliferation is the primary factor responsible for the increase in testis size and sperm production observed in teleost fish. In mammals, Sertoli cell proliferation in the adult testis is not observed under natural conditions. However, on the level of the individual spermatogenic cyst--similar to mammals--Sertoli cell proliferation ceases when germ cells have entered meiosis and when tight junctions are established between Sertoli cells. This suggests that fish are valid vertebrate models for studying Sertoli cell physiology.     

What makes a host profitable? Parasites balance host nutritive resources against immunity

Numerous host qualities can modulate parasite fitness, and among these, host nutritive resources and immunity are of prime importance. Indeed, parasite fitness increases with the amount of nutritive resources extracted from the host body and decreases with host immune response. To maximize fitness, parasites have therefore to balance these two host components. Yet, because host nutritive resources and immunity both increase with host body condition, it is unclear whether parasites perform better on hosts in prime, intermediate, or poor condition. We investigated blood meal size and survival of the ectoparasitic louse fly Crataerina melbae in relation to body condition and cutaneous immune response of their Alpine swift (Apus melba) nestling hosts. Louse flies took a smaller blood meal and lived a shorter period of time when feeding on nestlings that were experimentally food deprived or had their cutaneous immune response boosted with methionine. Consistent with these results, louse fly survival was the highest when feeding on nonexperimental nestlings in intermediate body condition. Our findings emphasize that although hosts in poor condition had a reduced immunocompetence, parasites may have avoided them because individuals in poor condition did not provide adequate resources. These findings highlight the fact that giving host immunocompetence primary consideration can result in a biased appraisal of host-parasite interactions.     

Small-sized euryhaline fish as intermediate hosts of the digenetic trematodeCryptocotyle concavum

Cercariae of the trematodeCryptocotyle concavum, which encyst in skin and/or kidney of sticklebacks and gobies, were studied in the Schlei Fjord (western Baltic Sea). Mean incidence of dermal cysts was 48 % inGasterosteus aculeatus and 37 % inPungitius pungitius. No cysts were found in the kidneys of sticklebacks. While 97 % ofPomatoschistus microps had encysted metacercariae in the kidneys, only 2 % had cysts in the skin.Pomatoschistus minutus, however, showed hardly any cyst infestation of either skin or kidney. InP. microps the intensity of infestation by metacercariae was frequently more than 50 cysts; in contrast, sticklebacks rarely exhibited more than 5 dermal cysts. Infested fish were larger than 10 mm in total length, the incidence rate increasing with growth. Parasitic infestation depends on ambient salinity:C. concavum was not found at salinities below 4 . In contrast to the high incidence in fish, the first hosts — the snailsHydrobia stagnalis andH. neglecta — showed remarkably low infection rates (3 to 5 %). The findings reported are related to the distribution ofC. concavum, the mode of life of infested fish, the feeding habits of the final hosts and the infestation ofP. microps by other parasites. Evidently,P. microps represents an optimal second host forC. concavum.     

Regulation of Reproductive Function in DrosophilaFemales Due to Hormonal Interaction Under Stress Is Genetically Determined

The effect of heat stress (38°C) on the content of octopamine (OA) and 20-hydroxyecdysone (20HE) was studied under normal and stressful conditions in adult flies of Drosophila virilislines contrasting in the level of the juvenile hormone (JH). The wild-type flies (line 101) exhibited a pronounced sex dimorphism for the content of both OA and 20HE, which was substantially lower in this line than in flies of the mutant line 147. The level of both hormones increased in flies of line 101 exposed to heat stress, whereas it remained unchanged in flies of line 147 under the same conditions. The effect of heat stress on the level of JH metabolism and fertility was also studied in D. melanogasterwild-type lines and lines carrying mutations in genes responsible for OA and DA syntheses. In octopamineless females of the Th ^nM18line and in females of the Steline characterized by a doubled content of DA, JH degradation differed from normal: it was increased in both young and mature Th ^nM18females, while decreased in young and increased in mature Steflies. Fertility was substantially lower in the Stethan in the wild-type line. Flies of all of the D. melanogasterlines produced a stress response; however, in mutant lines, both fertility and stress reactivity of the systems controlling JH metabolism differed significantly from that of the wild-type lines. The role of JH, 20HE, OA, and DA interaction in regulation of Drosophilareproduction under stressful conditions is discussed.     

Aspects of life-table studies and functional response of Lysiphlebia mirzai

The fecundity, reproductive rate, and survival of Lysiphlebia mirzai parasitising third instar nymphs of the cereal aphid Rhopalosiphum maidis were measured at six different host densities under constant laboratory conditions. The survival rate (l_x) of the female parasitoids was unaffected by host density, with an average adult life-span of 5–6 days at all densities. The age-specific fecundity rate (m_x) was host density-dependent. The value of m_x decreased rapidly from the first day of parasitisation. The number of hosts available determined the maximum possible number of mummies. At 200 hosts available per day, the average fecundity was 184.6 mummies/female; the maximum number of mummies yielded by any female was 200. The relationship between host density and the number of aphids parasitised per female was linear at 50 aphids/cage/day, but at higher host densities (100 aphids/cage/day) a significant curvilinear regression was observed. The intrinsic rate of natural increase (r_m) increased with increasing host density. Maximum value of r_m (0.262) was obtained at a host density of 200. The response of r_m to changes in host density and parasitoid sex ratio is shown. A typical type II functional response was observed for L. mirzai. The curve was described by a logistic curve, N_p=200/[1+exp(5.65–1.60 ln N_o)]. The search rate of the parasitoid was inverse host density-dependent. No significant variation in the sex ratio of F₁ offspring was observed at different initial host densities. Sex ratio values exceeded 0.5 at all host densities. The results evaluated the reproductive potential of L. mirzai as a promising biological control agent.     

Disease and the extended phenotype: parasites control host performance and survival through induced changes in body plan

Background

By definition, parasites harm their hosts. However, some forms of parasite-induced alterations increase parasite transmission between hosts, such that manipulated hosts can be considered extensions of the parasite''s phenotype. While well accepted in principle, surprisingly few studies have quantified how parasite manipulations alter host performance and survival under field and laboratory conditions.

Methodology/Principal Findings

By interfering with limb development, the trematode Ribeiroia ondatrae causes particularly severe morphological alterations within amphibian hosts that provide an ideal system to evaluate parasite-induced changes in phenotype. Here, we coupled laboratory performance trials with a capture-mark-recapture study of 1388 Pacific chorus frogs (Pseudacris regilla) to quantify the effects of parasite-induced malformations on host locomotion, foraging, and survival. Malformations, which affected ∼50% of metamorphosing frogs in nature, caused dramatic reductions in all measures of organismal function. Malformed frogs exhibited significantly shorter jumping distances (41% reduction), slower swimming speeds (37% reduction), reduced endurance (66% reduction), and lower foraging success relative to infected hosts without malformations. Furthermore, while normal and malformed individuals had comparable survival within predator-free exclosures, deformed frogs in natural populations had 22% lower biweekly survival than normal frogs and rarely recruited to the adult population over a two-year period.

Conclusions/Significance

Our results highlight the ability of parasites to deeply alter multiple dimensions of host phenotype with important consequences for performance and survival. These patterns were best explained by malformation status, rather than infection per se, helping to decouple the direct and indirect effects of parasitism on host fitness.     

Oviposition,larval preference,and larval performance in two polyphagous species: does the larva know best?

It is expected that females preferentially oviposit on plant hosts that allow for optimal larval performance. However, this expectation contradicts empirical evidence where adults do not always choose the best host for their descendants. Recent evidence suggests that females’ host selection depends on the number of potential hosts. Females from oligophagous species seem to be able to choose an appropriate host in terms of larval performance, whereas in polyphagous species, adult oviposition preference is not related with larval performance. This suggests that larvae in polyphagous species could be taking a more active role in host selection than their mothers. Here, we evaluated the oviposition preference and the larval preference and performance of two polyphagous species of economic importance, Copitarsia decolora (Guenée) (Lepidoptera: Noctuidae: Cuculliinae) and Peridroma saucia (Hübner) (Lepidoptera: Noctuidae: Noctuinae), on eight species of cultivated plants. In laboratory and greenhouse choice assays, we tested adult preference for oviposition and larval preference at 1 and 24 h. Larval performance was measured in terms of survival to adulthood, length of larval period, and pupal weight. We found that both adult females and larvae actively choose their hosts and that the larval preference toward the hosts is related to the females’ preference in both herbivore species. However, the females and larvae did not preferentially select the host with the best larval performance, indicating that larval performance is not related to female or larval preference and that other selective pressures are influencing the choice of the host plant in these two species.     

Analyse der physiologischen Bedingungen des Vermehrungswachstums männlicher Keimzellen von Drosophila melanogaster

Summary 48 h old testis were implanted into different adult hosts. In virgin females the germ cell multiplication is lower than in both mated females or in males. However in all host types the differentiation into spermatids beginns at the same time as in situ.The role of the different adult host glands in male germ cell development was investigated. Through inactivation of such glands in mated females or implantation of extra glands in virgin hosts was demonstrated, that not the adult Corpus allatum but the Median Neurosecretory Brain Cells (MNBC) are responsable for this germ cell multiplication.Adult host provided with larval Ring gland or pupal host accelerate the germ cell differentiation but not the germ cell multiplication of implanted testis. This acceleration would be much higher if the normal development of the germ cells in adult hosts were due to rests of pupal hormons in this milieu.The nutritive conditions of the host play a very important role in germ cell multiplication. The testis implanted in castrated females or females injected with casein hydrolisate show a higher number of developing germ cells. The MNBC controll the protein metabolism (assimilation) but are only indirectly responsable for the germ cell multiplication; the multiplication is a consequence of the autonomous protein synthesis of the germ cells themself.The growth of other imaginal discs in adult hosts are discussed in the light of these results.

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Herrn Prof. Dr. E.Hadorn bin ich zu Dank verpflichtet für die kritische Diskussion dieser Arbeit, Herrn Dr. R.Nöthiger und Herrn W.Gehring für die Hilfe bei der Abfassung des deutschen Textes.     

Comparative biology of two rosette crown-feeding flies of the genus Botanophila (Dipt., Anthomyiidae) with potential for biological control of their thistle hosts

The comparative biology of two anthomyiid flies with potential for biological control of their host thistles, Botanophila turcica on Carthamus lanatus and Botanophila spinosa on Onopordum acanthium , was studied using field surveys and collections, and by rearing collected eggs and larvae in the laboratory in southern France. The thistle hosts are significant weeds outside their native range, particularly in Australia. Both flies attack the rosette meristems of their hosts prior to flowering. Larval stages are described together with natural attack rates (21–33% of field plants) and the mortality of field-collected larvae reared in the laboratory. A successful rearing protocol for these flies is also described. The results of preliminary host-specificity tests showed that both species are highly specific, being restricted to their host genus in the tests conducted. Furthermore, Botanophila turcica could not complete development on safflower (a congener of its natural host) under natural conditions. Botanophila turcica had an adult activity period that lasted 6 months from late autumn and laid fewer eggs per host plant than B. spinosa , which was active for 2 to 3 months in spring. Only one parasitoid was observed, an endoparasitic eucoilid, Trybliographa sp., which attacked both fly species, causing 18–23% mortality.