Phortica variegata as an intermediate host of Thelazia callipaeda under natural conditions: evidence for pathogen transmission by a male arthropod vector

Knowledge about Phortica variegata (Drosophilidae, Steganinae), the intermediate host of the eyeworm Thelazia callipaeda (Spirurida, Thelaziidae), is confined to experimental studies. To investigate the role P. variegata plays in the transmission of T. callipaeda under natural conditions, the population dynamics of these flies in the natural environment and their feeding preferences (on vegetables and/or animal lachrymal secretions) were examined. From April to November 2005, a total number of 969 (557 males and 412 females) P. variegata flies were collected weekly in a region of southern Italy with a history of canine thelaziosis. The flies were identified and dissected or subjected to a PCR assay specific for a region within the ribosomal ITS-1 DNA of T. callipaeda. The zoophilic preferences of P. variegata were assessed by collecting flies around the eyes of a person or around a fruit bait. Seven hundred and twenty flies (398 males and 322 females) were dissected under a stereomicroscope; 249 flies (158 males and 91 females) that died prior to the dissection were subjected to molecular investigation. Only P. variegata males were infected with larval T. callipaeda both at dissection (six, 0.83%) and with the specific PCR (seven, 2.81%), representing a total percentage of 1.34% flies infected. Interestingly, only males were collected around the eyes, compared with a male/female ratio of 1:4 around the fruit. This survey indicated that P. variegata males act as intermediate hosts of T. callipaeda under natural conditions in Europe. Both the zoophilic behaviour of P. variegata males on lachrymal secretions and their role as vector of T. callipaeda have been discussed as they represent a peculiarity in medical and veterinary entomology. The synchrony between the fly population dynamics and the biology of the nematode in the definitive host provides an interesting model for exploring the co-evolution of Thelazia spp. with their hosts.     

Molecular characterization and phylogenesis of Steganinae (Diptera, Drosophilidae) inferred by the mitochondrial cytochrome c oxidase subunit 1

The subfamily Steganinae (Diptera, Drosophilidae) includes flies which display zoophilic feeding behaviour in the larval and/or adult stages, some of which act as vectors of Spirurida eyeworms, which infect both carnivores and humans. To date, the taxonomy and phylogeny of the subfamily Steganinae has been studied only superficially and many aspects of their systematics remain unresolved. Thus, the present study aimed to provide a molecular dataset to facilitate the identification and phylogenetic analysis of Steganinae species based on partial ( approximately 700 basepairs) mitochondrial cytochrome c oxidase subunit 1 (cox1) sequences. A total of 134 flies belonging to 13 species and eight genera of Steganinae were subjected to molecular and phylogenetic analyses. The mean nucleotide variation within the Steganinae subfamily was 8.1%, with a variation within genera for which more than one species was examined ranging from 1.6% (in Phortica spp.) to 21.8% (in Amiota spp.). Interspecific pairwise divergence ranged from 1.6% (Phortica variegata vs. Phortica semivirgo) to 24.8% (Cacoxenus indagator vs. Amiota alboguttata) and intraspecific variation ranged from 0% to 1%. Seventy of the 233 amino acids were variable, including 26 parsimony informative sites and 44 singleton sites, with some highly conserved residues identified within the genera Stegana and Amiota. Parsimony and maximum likelihood-based phylogenetic analyses provided strong support for the genus Phortica, phylogenetically distinct from the genus Amiota. Gitona distigma was placed in an unresolved position adjacent to the outgroup taxa, Drosophila yakuba and Drosophila melanogaster. The molecular data reported here represent the first molecular dataset based on cox1 of Steganinae flies and provide a base for further investigations into the evolutionary relationships among this little-studied subfamily.     

Factors affecting size, longevity and fecundity in the reindeer oestrid flies Hypoderma tarandi (L.) and Cephenemyia trompe (Modeer)

1. Laboratory reared reindeer oestrid flies Hypoderma tarandi and Cephenemyia trompe (Diptera: Oestridae) were weighed to determine progressive weight loss and death weights at treatments with various temperature and humidity conditions.
2. Four individual measurements of size were taken: larval weight, wet weight of newly eclosed flies, wing length, and weight of flies after dehydration and fat extraction. In H. tarandi, males were bigger than females (except for wing length), whereas the reverse was true for C. trompe .
3. Size variation was not significantly related to conditions (temperature, humidity, duration) during the pupal stage, but individual reindeer produced flies (both species) of different mean sizes. These size differences were not correlated with larval burden (= number of larvae per individual host), but are hypothesized to be connected to unknown host quality factors.
4. Longevity of flies kept in vials and subjected to various temperature and humidity conditions revealed that C. trompe lived significantly longer than H. tarandi (range: 4–44 and 1.2–27 days, respectively) at 5–33 °C. Male H. tarandi survived longer than females; female C. trompe survived longer than males. Longevity was not significantly correlated to any of the size measures.
5. Most flies had a large portion of their fat reserves left at death.
6. In H. tarandi , mean number of eggs was 609 ± SD 73 (range 354–772, n = 119). Egg number was slightly dependent on larval size, but not on wet weight of newly eclosed flies or wing length. In C. trompe , mean number of eggs was 960 ± SD 208 (range 493–1349, n = 31).
7. The possible adaptive value of large size in oestrids is questioned. Benefits of flexibility in size in oestrids are hypothesized.     

Elemental stoichiometry of Drosophila and their hosts

1. Nitrogen (N) and phosphorus (P) availabilities are important ecological determinants of resource use in nature. Despite the wide range of hosts used by species of the genus Drosophila , elemental composition of natural resources of these flies has never been investigated.
2. Total body N and P contents were determined in seven species of wild-caught Drosophila , their natural hosts, and artificial diets routinely used to rear these flies in the laboratory. The flies tested included D. hydei, D. arizonae, D. simulans and D. pseudoobscura collected from rotting fruit (melons), and the cactophilic D. nigrospiracula, D. mojavensis and D. pachea collected from their specific host plants, Saguaro, Organpipe and Senita cactus, respectively.
3. Natural hosts varied in elemental composition, with fruit showing higher N (2·8–4·3% dry mass) and P (0·50–0·67%) levels compared with cacti (0·5–1·6% N; 0·01–0·29% P). No consistent differences in N and P levels were found between healthy and necrotic cactus tissue.
4. Total body N and P also varied among Drosophila species. This variation mirrored the levels of N and P found in the respective hosts and laboratory diets. N:P ratios were consistently lower in female flies compared with conspecific males suggesting phosphorus demands during oogenesis are high.
5. Potential mechanisms by which Drosophila deal with N or P limitation in nature are discussed.     

Diversity,ecology, and seasonality of sand flies (Diptera: Psychodidae) of the Jenin District (Palestinian Territories)

The diversity, ecology, and seasonality for sand flies from two localities in Jenin District, the Palestinian Territories, were studied. A total of 12,579 sand flies (5,420 Phlebotomus and 7,159 Sergentomyia) were collected during the study period. The genera Phlebotomus and Sergentomyia are represented by 13 and nine species and subspecies, respectively. Species account was given for all collected species. CDC light traps yielded 7,649 (60.8%) of the total captured sand flies, while sticky traps and aspirators contributed to 36.4 and 2.8% of the total collected specimens, respectively. Phlebotomus sergenti and P. syriacus showed two peaks, one in July and one in October. Phlebotomus tobbi showed one peak towards the end of the summer in September and August, while P. papatasi showed a bimodal peaks pattern, one in June and one in October. Phlebotomus canaaniticus showed a peak in August. P. perfiliewi transcaucasicus and P. neglectus showed a peak in October. Sergentomyia dentata showed one peak in August and increasing numbers from June to August, declining afterwards. Other species, such as S. theodori, had one peak in June, S. taizi had steady numbers across the summer, and S. christophersi had a peak in August.     

Egg size, first instar behaviour and the ecology of Lepidoptera

There is striking variation in egg size among Lepidoptera. Part of the explanation could be a link between egg size and larval feeding ecology.
The relationship between absolute egg size and aspects of feeding ecology for different Lepidoptera families from different temperate regions is examined. Species that overwinter in the egg stage have larger eggs. There are significant differences in egg size with respect to feeding specificity but different families show different patterns. Woody plant feeders have larger eggs than herb feeders. There is little effect of proximity of the egg to the plant part that is eaten.
Patterns in the behaviour and survival of newly hatched larvae of 42 spp. of British Lepidoptera and their relationship to egg and larval size and to food plant characteristics are examined. Patterns in egg size with respect to feeding ecology are similar to those described above. There is a strong correlation between egg size and the size of newly hatched larvae. Newly hatched larvae survived for a mean of 1–20 days without food. Survival is not correlated with larval weight. Grass feeders survive longer than herb and woody plant feeders; the species surviving the longest feeds on lichens. Newly hatched larvae moved at a mean speed of 0.7-267.8 cm h^-1. Speed is not correlated with larval weight or survival time. Grass feeders move faster than woody plant feeders which, in turn, move faster than herb feeders. Woody plant feeders tend to move upwards, grass feeders downwards and herb feeders both upwards and downwards. The proportion of larvae silking is negatively correlated with larval weight.
The strong links between egg size and larval feeding ecology and between feeding ecology and larval behaviour are discussed. It is surprising that larval body size does not appear to constrain the speed of movement, nor tolerance to starvation.     

Reproductive characteristics of the flower breeding Drosophila hibisci Bock (Drosophilidae) along a latitudinal gradient in eastern Australia: relation to flower and habitat features

For many insect species, egg and larval substrate characteristics are significantly correlated with interspecific differences in female reproductive allocation and egg size-number tradeoffs. We tested the hypothesis that a similar pattern occurred within the Australian drosophilid, Drosophila hibisci, that is restricted throughout its life cycle to flowers of species in the genus Hibiscus. These plants occur as small, isolated, normally monospecific stands that should facilitate differentiation of the fly populations in relation to specific oviposition and larval substrates. Data from 38 sites ranging from 20.8̀ to 34.4̀ S latitude in eastern Australia indicated no relationship between female body size, egg size, or ovariole numbers and floral size or mass among four species of Hibiscus. However, the flies did show a latitudinal cline in ovariole number that was independent of floral variation. Females averaged 15–20 ovarioles per female in the south (32–34̀ S latitude) and 10–12 ovarioles in the north (21–22̀ S latitude). The increase in ovariole number with latitude was due to divergence in the ovariole number of the largest females. In contrast, small females in the north and south had the same number of ovarioles. Reproductive allocation of female flies in the northern region was less than females in the southern region. The latitudinal divergence in ovariole number was not associated with habitat differences (density of trees, density of flies and beetles), nor with differences in floral characteristics (flower weight, petal length, yeast species present). Short term weather patterns in daily temperature and rainfall preceding collections pardy explain the variation in ovariole number. These observations in conjunction with preliminary genetic results suggest the cline is associated with genetic differences that interact with environmental determinants such as the temperature during larval development.     

Vertical distribution of haematophagous Diptera in temperate forests of the southeastern U.S.A.

The vertical distribution of blood‐feeding flies in two temperate forests in the southeastern U.S.A. was determined by placing 15 Centers for Disease Control and Prevention miniature light traps (12 CO₂‐baited, three unbaited controls), without lights, at three heights (1.5 m, 5.0 m, 10.0 m). More than 6550 haematophagous flies, representing 49 species in four families, were collected. Eighteen species were taken almost exclusively (90–100%) at 1.5 m or 10.0 m, and the mean number of flies per trap differed significantly with height for another six species. Five species exhibited shifts in vertical distribution between the two forests, indicating that forest structure could influence the height of host searching. Most (52.5%) mammalophilic flies were collected at 1.5 m, whereas most (56.4%) ornithophilic flies were taken at 10.0 m, suggesting that host associations influence vertical distributions. The significant differences in the composition of haematophagous fly populations among forest strata emphasize the importance of trap placement in vector surveillance and of understanding the ecological relationships of blood‐feeding flies.     

Larval habitats of sand flies in rural areas of southern Brazil

We report the results of an investigation of natural larval sand fly habitats in the Recanto Marista, Doutor Camargo municipality, Paraná state, Brazil, from May, 2010 to August, 2012. We used Alencar emergence traps (AT), experimental traps (ET), and soil samples incubated in a biochemical oxygen demand incubator. Eight sand flies were collected with ATs. One specimen was collected with an ET and 21 were collected in soil samples. The collected species were Brumptomyia brumpti, Micropygomyia ferreirana, Migonemyia bursiformis, Migonemyia migonei, Nyssomyia neivai, Nyssomyia whitmani, and Pintomyia pessoai. The laval habitats of sand flies were located in the Recanto Marista, especially between tree roots, but the number of adults that emerged in the traps and soil samples was small despite the high density of sand flies that has been recorded in the Recanto Marista.     

Notes on the biology and ecology of the parasitoids of the poplar clearwing moth, Paranthrene tabaniformis (Rott.) (Lep., Sesiidae) in Bulgaria. II. Eriborus terebrans (Gravenhorst, 1826) (Hym., Ichneumonidae)

The biology and ecology of Eriborus terebrans (Grav.), a parasitoid of the poplar clearwing moth, Paranthrene tabaniformis (Rott.), were studied during the period 1987–98. One-year-old poplar ( Populus spp.) shoots infested with P. tabaniformis larvae were collected from poplar seedlings at 11 localities in Bulgaria and examined in both field and laboratory conditions. Eriborus terebrans was recorded in seven localities as a solitary internal larval parasitoid of P. tabaniformis which developed two generations in early and mid-stage host larvae. Eriborus terebrans overwintered as a larva in P. tabaniformis overwintering larvae. In the field adult parasitoids of the overwintering generation appeared between late April/early May, and June or July. The peak activity of E. terebrans adults only coincided with the beginning of host emergence, which resulted in low levels of parasitism, being no more than 6.2%. Parasitoid adults of the summer generation appeared in late June–mid August. In this period enough larvae of the host were suitable for attacking and parasitism reached 24.4–39% in some cases. The average mortality of P. tabaniformis overwintering larvae caused by this parasitoid in Bulgaria during the period of the study was 4.7%. A significant part of the parasitized P. tabaniformis larvae constructed tunnel structures of frass and silk threads over the external openings of the galleries. It is possible that these structures protect the parasitoid cocoons from natural enemies – hyperparasitoids and predators.