Maggots cannot live on meat meal alone: production parameters for mass rearing of the ovoviviparous blowfly, Calliphora dubia (Diptera: Calliphoridae).

This study determined a cost-effective larval diet for rearing Calliphora dubia Macquart for use as a potential managed pollinator in Australia. This fly has potential as a pollination species to support honey bees (Apis mellifera). Larvae of C. dubia were reared mostly in meat meals with varying amounts of either whole egg powder, whole eggs (+ shell), bran flakes, skimmed milk powder, brewer's yeast, or poultry oil. This was done from an economic and production perspective to support commercial rearing. Several laboratory-based studies determined the growth and output from various ingredient combinations. Larvae fed 90% meat meal and 10% whole egg powder developed rapidly through to pupation with a high pupation rate, adult size, and percent adult emergence. Given the high cost and difficulty in sourcing whole egg powder, media comprising mostly meat meals with the addition of bran flakes and whole eggs also supported rapid larval development, pupation rate, and adult emergence. The ideal amount of media/larvae was 0.5 g/larvae to support high pupation rates and adult emergence. Adult eclosion occurred over 4-5 days, even when larvae were laid and fed within 1 h on ample media. Commercial mass rearing would then require daily cohorts of larvae to ensure peak adult fly emergence over 1-2 days for release into a crop. Mass-rearing C. dubia should use meat meal as the base ingredient with bran flakes and whole eggs added and fed at 0.5 g of media/larvae. Based on the current media ingredient costs, rearing 1-m adult C. dubia would cost just over $500 (US$342).

Implications of Seizure-Cluster Treatment on Healthcare Utilization: Use of Approved Rescue Medications.

Purpose: People with epilepsy may experience seizure clusters despite a stable regimen of antiseizure medications. Such clusters have the potential to last ≥24 hours, typically occur in the community setting, and may progress to medical emergencies, such as status epilepticus, if untreated. Thus, long-acting rescue therapy for seizure clusters is needed that can be administered by nonmedical individuals outside a hospital. Benzodiazepines are the foundation of rescue therapy for seizure clusters. The approved outpatient treatments (ie, diazepam, midazolam) have differing profiles that may affect multiple aspects of health-care utilization. The current labeling of these medications allows for a second dose if needed to control the cluster. Although no head-to-head studies directly comparing rescue treatments have been conducted, differences between studies with generally similar designs may provide context for the potential importance of second doses of rescue therapy on health-care utilization. Methods: For this analysis, large, long-term, open-label studies of approved seizure-cluster treatments designed for use by nonmedical caregivers were reviewed, and the percentage of seizure clusters for which a second dose was used or that were not controlled at 6, 12, and 24 hours was examined. Available data on hospitalizations were also collected. Results: The 3 identified studies meeting the inclusion criteria were for use of diazepam rectal gel, intranasal midazolam, and diazepam nasal spray. Across these studies, the use of a second dose ranged from <40% at 6 hours to <13% at 24 hours. Hospitalizations and serious treatment-emergent adverse events were reported variably across these studies. Conclusion: These results demonstrate the importance of second doses of rescue therapy for seizure clusters for optimizing health-care utilization. Need for second doses should be included as one component. In turn, when second doses are needed, they have the potential to curtail emergency department use and hospitalization and to prevent further seizure clusters.

Examining the patient and caregiver experience with diazepam nasal spray for seizure clusters: Results from an exit survey of a phase 3, open-label, repeat-dose safety study.

BACKGROUND: Ideal rescue treatments for acute treatment of seizure clusters should be easy to administer, so it is important to assess user perceptions of these treatments. Diazepam nasal spray is designed to have a rapid, noninvasive, and socially acceptable route of administration. Patient and caregiver (including care partner) responses to surveys from a phase 3 safety study of diazepam nasal spray are reported. METHODS: The study enrolled patients aged 6-65 years with seizure clusters. Surveys distributed to patients and caregivers at study end, completion, or discontinuation collected data on comfort using diazepam nasal spray outside the home, timing of administration and return to their usual selves, and comfort of use compared with rectal diazepam. Safety was assessed. RESULTS: Of 175 patients enrolled at the October 31, 2019, interim cutoff, 158 received diazepam nasal spray. Sixty-seven (42.4%) patients and 84 (53.2%) caregivers responded to the surveys (including 35 matched pairs). Most patients (78.8%, 52/66) responded that they were very comfortable doing activities outside the home with diazepam nasal spray available; 59.4% of patients returned to their usual selves within an hour of administration. Twenty-seven (40.3%) of these patients reported self-administration, 48% doing so at the first sign of a seizure. Administration of diazepam nasal spray was rated extremely or very easy by 93.8% of caregivers. Safety profile was consistent with diazepam rectal gel; no patient discontinued owing to treatment-emergent adverse events. Nasal discomfort was typically mild and transient. Among patients who had used diazepam rectal gel, most were not at all comfortable using it outside the home (86.7%) or at home (64.5%) compared with diazepam nasal spray, whereas caregivers reported that diazepam rectal gel was not at all easy to use compared with diazepam nasal spray. CONCLUSIONS: This survey from the phase 3 safety study of diazepam nasal spray shows that patients and caregivers were satisfied with, and more comfortable using, diazepam nasal spray than rectal diazepam in public. NCT02721069.

Evaluation of diazepam nasal spray in patients with epilepsy concomitantly using maintenance benzodiazepines: An interim subgroup analysis from a phase 3, long-term, open-label safety study.

OBJECTIVE: Diazepam nasal spray (Valtoco), indicated for acute treatment of frequent seizure activity (seizure clusters) in patients with epilepsy ≥6 years of age, is designed to be a rapid, noninvasive, socially acceptable route of administration. This interim analysis evaluated the safety profile of diazepam nasal spray in patients with and without concomitant use of benzodiazepines, with use of a second dose for a seizure cluster as a proxy for effectiveness. METHODS: A long-term, phase 3, open-label safety study enrolled patients with epilepsy who had seizures despite a stable antiseizure medication regimen. RESULTS: Among 175 patients enrolled by October 31, 2019, a total of 158 were treated with diazepam nasal spray (aged 6-65 years; 53.8% female). Of those, 119 (75.3%) received concomitant benzodiazepines (60, chronic; 59, intermittent); 39 (24.7%) did not. Use of a second dose was similar in patients using chronic concomitant benzodiazepines (second dose in 11.1% [144/1299]) and those with no concomitant benzodiazepines (second dose in 10.3% [41/398]). Treatment emergent adverse events (TEAEs) occurred for 80.0% with chronic use of concomitant benzodiazepines and 61.5% without. Cardiorespiratory depression was not reported, and no serious TEAEs were treatment related. Study retention was high: 83.3% in the chronic benzodiazepine group and 76.9% in the no-benzodiazepine group. Findings were similar in a sub-analysis of patients who were (n = 44) or were not (n = 75) taking clobazam. SIGNIFICANCE: This analysis of patients from a long-term study shows a similar safety profile of diazepam nasal spray in patients with and without concomitant benzodiazepines, and consistent with the established profile for diazepam. Use of a single dose of diazepam nasal spray and high study retention rates suggest the effectiveness of diazepam nasal spray in patients irrespective of chronic daily benzodiazepine use. Results were similar in the clobazam sub-analysis. These results support the safety and effectiveness of diazepam nasal spray in patients with concomitant benzodiazepine use.

The Role of Flies as Pollinators of Horticultural Crops: An Australian Case Study with Worldwide Relevance.

Australian horticulture relies heavily on the introduced managed honey bee, Apis mellifera Linnaeus 1758 (Hymenoptera: Apidae), to pollinate crops. Given the risks associated with reliance upon a single species, it would be prudent to identify other taxa that could be managed to provide crop pollination services. We reviewed the literature relating to the distribution, efficiency and management potential of a number of flies (Diptera) known to visit pollinator-dependent crops in Australia and worldwide. Applying this information, we identified the taxa most suitable to play a greater role as managed pollinators in Australian crops. Of the taxa reviewed, flower visitation by representatives from the dipteran families Calliphoridae, Rhiniidae and Syrphidae was frequently reported in the literature. While data available are limited, there was clear evidence of pollination by these flies in a range of crops. A review of fly morphology, foraging behaviour and physiology revealed considerable potential for their development as managed pollinators, either alone or to augment honey bee services. Considering existing pollination evidence, along with the distribution, morphology, behaviour and life history traits of introduced and endemic species, 11 calliphorid, two rhiniid and seven syrphid species were identified as candidates with high potential for use in Australian managed pollination services. Research directions for the comprehensive assessment of the pollination abilities of the identified taxa to facilitate their development as a pollination service are described. This triage approach to identifying species with high potential to become significant managed pollinators at local or regional levels is clearly widely applicable to other countries and taxa.

Amending Poultry Broiler Litter to Prevent the Development of Stable Fly, Stomoxys calcitrans (Diptera: Muscidae) and Other Nuisance Flies.

Spent poultry litter use as a fertilizer in horticulture supports stable fly Stomoxys calcitrans (L.) (Diptera: Muscidae) development. Stable fly continues to have an economic impact on livestock production and rural lifestyle in south-western Australia. The use of raw poultry manure is banned in 12 Shires surrounding Perth. The loss of market options for West Australian broiler growers has caused economic hardship. Hence, this study examined a range of chemical and biological amendments to spent poultry broiler litter in preventing stable fly and nuisance fly development. These included alkalizers (i.e., lime sand, quicklime, soda ash, and shell grit), acidifiers (aluminum sulfate, sodium bisulfate), gypsum, zeolite, spongolite, calcium cyanamide, and two fungal agents. The treated litters were placed under irrigation in horticulture with amendments added prior to them being exposed in the field as replicate 1-liter pads. In total, 19,559 stable flies developed from the spent litters exposed over five field experiments (88.7% of all flies recovered). House flies (Musca domestica L. (Diptera: Muscidae); 2,067 or 9.4%), false stable flies (Muscina stabulans Fallén (Diptera: Muscidae); 414 or 1.9%), and two sarcophagids (flesh fly) also developed from the litter. Borax completely prevented any fly development from the litter. Calcium cyanamide (1-2.5% v/v) and sodium bisulfate (10%) reduced stable fly numbers by as much as 99-100% when added to litter. Alkalizers, zeolite, spongolite, and entomopathogenic fungi had no significant impact on stable fly development. The addition of either calcium cyanamide or sodium bisulfate to raw litter can boost the fertilizer value of the litter while preventing stable fly development.

Survival and development of the forensically important blow fly, Calliphora varifrons (Diptera: Calliphoridae) at constant temperatures.

The influence of temperature on the development of the forensically important blow fly, Calliphora varifrons Malloch 1932 (Diptera: Calliphoridae), was investigated at seven constant temperatures ranging from 12 to 30 °C. C. varifrons completed development between 12 and 27 °C. At 30 °C larvae formed pupae but did not successfully emerge. Temperature significantly influenced development time, mortality, maximum larval length, and adult body size. Development time (larviposition to adult emergence) ranged from 16.65 ± 0.17 days at 27 °C to 49.93 ± 0.26 days at 12 °C. Development rate was essentially linear throughout the 12-27 °C temperature range. Linear estimates of lower developmental threshold and thermal requirement (K) for development of C. varifrons were 4.20 °C and 368.46 ± 26.38 K. At 30 °C, a slight inhibitory effect of high temperature on third instar development rate was observed followed by a rapid decline when subsequent development and survival ceased. Nonlinear estimates of lower developmental threshold for third instar development were comparatively higher (6.29 °C). Nonlinear estimates of optimal developmental temperature and upper lethal developmental threshold were 25.94 and 32.13 °C respectively. Mortality was high at both temperature extremes (12 and 27 °C) and lowest between 18 and 24 °C. Maximum larval length was inversely related to temperature. Adult body size was significantly smaller at 12 °C, peaked at 18 °C, and declined as temperatures increased. The species-specific development data presented are the first available for C. varifrons for use by forensic practitioners for estimation of minimum time since death.

The laying of live larvae by the blowfly Calliphora varifrons (Diptera: Calliphoridae).

This study examined larviposition in Calliphora varifrons Malloch (Diptera: Calliphoridae), an ovoviviparous blowfly of forensic importance in Western Australia. Females took 1.5 min (mean 89±10.0s, n=42) to deposit their live, first instar larvae (average body length 2.5 mm) on fresh liver in the laboratory. Females laid an average of 30 live larvae (mean of 29.9±1.2, n=50) at a rate of 1 larva every 2 s (mean of 0.46±0.05 larvae/s, n=42). Most females (66%) laid live larvae only, but 18% laid fully developed eggs (half of the egg masses laid by individual females were viable) and 16% laid live larvae and some eggs at the same time. Females laying only eggs laid a mean of 24.1±3.0, n=10 eggs on each occasion. Field females carried between 20 and 44 live larvae (mean of 33.4±1.0, n=49) with a strong linear relationship between female size and number of live larvae. In the laboratory, females typically laid larvae in multiple locations. Very few females (<8%) retained some larvae in their ovisacs, retaining on average, half of their complement of developed larvae. Those larvae retained had begun to be resorbed by the female with necrosis evident on the posterior segments.

Decomposition and insect succession of clothed and unclothed carcasses in Western Australia.

The effect of clothing on carcass decomposition and patterns of insect succession onto remains were investigated in two separate years during autumn in Western Australia. The progression of decomposition differed between clothed and unclothed carcasses in both years of the study. The presence of clothing markedly prolonged the wet decay stage in both years with larval feeding occurring across the moist skin surface underneath clothing, as well as within and under the carcasses. Ambient temperatures were higher in the second year of the study and corresponded to marginally faster rates of decay throughout decomposition. Within years, insect arrival and oviposition were largely consistent between clothed and unclothed carcasses with a few notable exceptions. The green blow fly, Lucilia sericata Meigen (Diptera: Calliphoridae) oviposited one day earlier on clothed than unclothed carcasses in both years of the study. The black carrion fly, Australophyra rostrata Robineau-Desvoidy, (Diptera: Muscidae) colonised clothed carcasses in two distinct waves of succession but only one wave of ovipoistion was observed on unclothed carcasses in either year. Correspondingly, clothed carcasses supported larval feeding by A. rostrata for a longer duration than unclothed carcasses. Finally, dipteran larval masses were more widely distributed across the carcass surface and were present for a longer period of time on clothed carcasses than on unclothed carcasses in both years. Forensically relevant data detailing the seasonal pattern of insect succession onto clothed and unclothed decomposing remains in Western Australia are reported.