Growth and survival of blowfly Lucilia sericata larvae under simulated wound conditions: implications for maggot debridement therapy.

Maggot debridement therapy has become a well-established method of wound debridement. Despite its success, little information is available about the optimum duration of the treatment cycle and larval growth in wounds. This study examines the development of Lucilia sericata (Diptera: Calliphoridae) larvae under two containment conditions (bagged and free range) under simulated wound conditions and assesses the impact of transport and further storage of larvae on their survival and growth. There was no significant difference in size between bagged and free-range larvae over the 72-h experimental period. Larvae grew fastest 8-24 h after inoculation and completed their growth at 40-48 h. Mortality rates were similar (0.12-0.23% per hour) in both containment conditions and did not differ significantly (P = 0.3212). Survival of free-range larvae was on average 16% lower than survival of bagged larvae. Refrigeration of larvae upon simulated delivery for > 1 day reduced their survival to < 50% and caused a reduction in growth of up to 30% at 12 h, but not at 48 h, of incubation. Therefore, it is recommended that free-range larvae are left in the wound for a maximum of 40-48 h, and bagged larvae for 48-72 h. Larvae should be used within 24 h of delivery to avoid high mortality caused by prolonged refrigeration.

The influence of housefly Musca domestica embryo age on viability in water and at low temperatures.

The sensitivity of housefly Musca domestica L. (Diptera: Muscidae) embryos to storage at low temperatures (5 and 10 °C on moist sponges in Petri dishes) and in water at 26 °C was investigated to develop suitable protocols for the storage and transport of housefly eggs. The youngest embryos (aged 0-3 h) were the most sensitive to storage at 5 °C, with 45% survival after storage for 24 h. Storage of embryos aged 3-12 h at 5 °C for 24 h had no negative effect; longer storage resulted in significantly decreased larval survival (30-34% after 48-72 h, compared with 61% in the control group) and reduced hatching rates (83% after 72 h storage). No negative effects were observed when embryos aged 0-9 h were stored at 10 °C for 24 h, but this temperature did not completely inhibit development and eggs began to hatch if stored for longer than 24 h. All age groups of embryos showed high mortality after storage in water at 26 °C for 24 h, with the youngest embryos being least resistant to submersion.

Improvement of survival of the house fly (Musca domestica L.) larvae under mass-rearing conditions.

Two new approaches were examined, aimed at increasing survival of the house fly (Musca domestica L.) larvae under mass-rearing conditions of a biodegradation facility: modification of the larval substrate and dispersal of the eggs during inoculation. The two types of pig manure used in this study (manure with sawdust and manure without sawdust) differed in terms of larval survival and nutritional value for the house fly larvae. Larval survival in manure without sawdust in the control treatment was low (46.8 ± 2.1%) and its nutritional value for the larvae were high. Addition of 5.7% of previously biodegraded manure did not significantly affect larval survival (52.3 ± 1.9%), but larval development was faster and the pupae were significantly smaller (14.28 ± 0.4 mg) compared to the control (16.29 ± 0.5 mg). Using alternative substrate for incubation of eggs and first-instar larvae significantly increased larval survival (63.3 ± 3.3%) and decreased the mean weight of produced pupae (14.39 ± 0.71 mg). Overall, the weight of recovered biomass in the alternative substrate treatment increased by 14.3 kg ton-1 of manure compared to the control. Larval survival in manure with sawdust was generally higher than 70%, but its nutritional value for the larvae was lower than in manure without sawdust. Dispersal of eggs over the surface of manure with sawdust significantly affected larval survival and mean weight of pupae. Larval survival was significantly lower (59.2 ± 4.0%) and pupae were significantly heavier (18.45 ± 0.8 mg) when eggs were applied to a small area on the manure surface (spot treatment), as compared to diagonal, Z-line and multiple zig-zag dispersal (72.5 ± 2.4 to 74.6 ± 3.0% and 14.76 ± 0.6 to 15.97 ± 0.6 mg, respectively). No significant differences were observed in larval survival or mean weight of pupae when comparing the diagonal, Z-line and multiple zig-zag dispersal patterns. Implementation of the techniques which improve larval survival and increase the weight of produced fly biomass may decrease demand for production of house fly eggs and, therefore, reduce the maintenance costs of adult colony, as well as increase the revenue earned by selling the products.

In-vivo time-dependent articular cartilage contact behavior of the tibiofemoral joint.

OBJECTIVE: The purpose of this study was to investigate the in-vivo time-dependent contact behavior of tibiofemoral cartilage of human subjects during the first 300 s after applying a constant full body weight loading and determine whether there are differences in cartilage contact responses between the medial and lateral compartments. DESIGN: Six healthy knees were investigated in this study. Each knee joint was subjected to full body weight loading and the in-vivo positions of the knee were captured by two orthogonal fluoroscopes during the first 300 s after applying the load. Three-dimensional models of the knee were created from MR images and used to reproduce the in-vivo knee positions recorded by the fluoroscopes. The time-dependent contact behavior of the cartilage was represented using the peak cartilage contact deformation and the cartilage contact area as functions of time under the constant full body weight. RESULTS: Both medial and lateral compartments showed a rapid increase in contact deformation and contact area during the first 20s of loading. After 50s of loading, the peak contact deformation values were 10.5+/-0.8% (medial) and 12.6+/-3.4% (lateral), and the contact areas were 223.9+/-14.8 mm(2) (medial) and 123.0+/-22.8 mm(2) (lateral). Thereafter, the peak cartilage contact deformation and contact area remained relatively constant. The respective changing rates of cartilage contact deformation were 1.4+/-0.9%/s (medial) and 3.1+/-2.5%/s (lateral); and of contact areas were 40.6+/-20.8 mm(2)/s (medial) and 24.0+/-11.4 mm(2)/s (lateral), at the first second of loading. Beyond 50 s, both changing rates approached zero. CONCLUSIONS: The peak cartilage contact deformation increased rapidly within the first 20s of loading and remained relatively constant after approximately 50 s of loading. The time-dependent response of cartilage contact behavior under constant full body weight loading was significantly different in the medial and lateral tibiofemoral compartments, with greater peak cartilage contact deformation on the lateral side and greater contact area on the medial side. These data can provide insight into normal in-vivo cartilage function and provide guidelines for the improvement of ex-vivo cartilage experiments and the validation of computational models that simulate human knee joint contact.

Maggot debridement therapy.

AIM: To establish a maggot debridement therapy unit and assess efficacy of this method in chronic conservative non-treatable leg ulcers in patient in whom conventional therapy failed. MATERIAL AND METHODS: From August 2004 to December 2005 we treated with Maggot debridement therapy (MDT) 10 patients, 6 men and 4 women, with 13 leg ulcers treated first with conventional and then with maggot therapy. Average age of these patients was 55 years, 11 wounds were of venous and 2 of arterial origin. 7 patients had diabetes mellitus. RESULTS: 1 patient with arterial ulcers underwent supracondylar amputation, 2 patients were re-admitted for recurrent ulcers, one extremely obese patient with cardiovascular co-morbidity did not heal, but all other ulcers have been healed, cleared or minimised during 4-8 week MDT. We observed massive growth of granulation tissue and microbiological cleaning of these wounds. No adverse effects were observed. CONCLUSION: Maggot therapy was more effective and efficient in debriding non-healing leg ulcers than a conservative treatment (Fig. 3, Ref. 6).

Anticoagulant activities in salivary glands of tabanid flies.

Tabanid flies are telmophages (pool feeders), taking frequent and rapid bloodmeals from many different individual hosts. To investigate how they accomplish this intermittent feeding strategy, we examined the anticoagulant activities in salivary gland extracts (SGE) from 19 species representing six genera: Atylotus, Chrysops, Haematopota, Heptatoma, Hybomitra and Tabanus (Diptera: Tabanidae). Standard coagulation screen assays were used to determine thrombin time, prothrombin time and activated partial thromboplastin time. Chromogenic substrate assays were performed for thrombin and factor Xa activity. SGE of most species (except Chrysops spp.) considerably prolonged human plasma clotting time in a dose-dependent manner, and showed potent and specific antithrombin activity in the chromogenic substrate assay. Heptatoma pellucens displayed the strongest anticoagulant activity. Specific anti-factor Xa activity in tabanid SGE was not detected. Electrophoretic profiles of SGE proteins differed between genera and species. Overall, the results suggest that tabanids have evolved at least two antihaemostatic strategies.

Identification of anticoagulant activities in salivary gland extracts of four horsefly species (Diptera, tabanidae).

Anticoagulant activities against the extrinsic and intrinsic coagulation pathways were identified in salivary gland extracts (SGE) prepared from four tabanids (Hybomitra muehlfeldi, Tabanus autumnalis, Haematopota pluvialis, Heptatoma pellucens). All extracts prolonged human plasma clotting time in a dose-dependent manner and inhibited thrombin activity in the chromogenic substrate assay. Horsefly SGE did not inhibit factor Xa. Partial purification of SGE proteins using reversed-phase high-performance liquid chromatography revealed species-specific differences in the elution profiles and range of fractions with anticoagulant activities.