The purpose of this study was to evaluate the post—embryonic development of Chrysomya putoria (Wiedemann 1818) (Diptera: Calliphoridae) reared on a diet of gizzard or gizzard/agar homogenate, with a diet of beef used as the control. Four replicates per treatment were performed (60 mL of each diet). The gizzard (60%), distilled water, and agar homogenate were combined in a blender. Each replicate consisted of 40 newly hatched larvae of C. putoria (5th generation). Each glass beaker containing a diet was inserted into a larger flask containing sawdust, which was covered with a nylon cloth held in place by an elastic band. The larvae were weighed and stored in test tubes sealed with a nylon cloth and an elastic band. The average temperature, measured with a thermohygrograph, was 20.6 °C, and the average relative humidity was 67.7%. The variation in the mean weight of mature larvae and in the duration of the larval, pupal, and total stages (newly hatched larvae to imagoes) were analyzed by Student's t—test (α = 5%), while viability was compared by ANOVA. The sex ratio was evaluated by the chi—squared test. The average duration of the period from the larval to imago stage was 8.868 days on the beef diet, 8.676 on the gizzard diet, and 9.067 on the gizzard/agar homogenate diet. Larval survival rates on these diets were 98, 92, and 73%, respectively, while pupal viabilities were 98, 91, and 71%, respectively, and larva—to—imago viabilities were 93, 83, and 64%, respectively. The duration of the pupal period differed significantly between the blowflies reared on the beef and gizzard/agar homogenate diets. The two diets proved to be good alternatives for rearing C. putoria.
Large-scale, quality-controlled laboratory production of fly larvae is needed for biotherapy. The objective of this study was to assess the action of glutaraldehyde on the sterilization of Chrysomya putoria eggs by applying pharmaceutical sterility tests. Egg masses with 0.600 g were divided into three parts of 0.200 g, the eggs were separated using sterile distilled water, and the suspensions obtained were mixed with activated 2% glutaraldehyde solution. After 15-min contact, the suspensions were filtered through Whatman filter paper, and the glutaraldehyde residue obtained in the filtrate was neutralized by rinsing with Tryptone Soy Broth. The treated eggs were placed aseptically on Petri dishes containing gauze moistened with sterile saline solution. About 10% of the sterilized mass was transferred to test tubes containing Tryptone Soy Broth and Fluid Thioglycollate Broth. The tubes were incubated, respectively, at 22.5 and 35.0°C for 14 d to verify egg mass sterility. The plates containing the rest of the eggs (90%) were sealed with plastic film and kept in a climatized chamber at 30°C/d, 28°C per night, 60 ± 10% relative humidity, and under a 12-h light period to assess insect viability and survival. Each experiment was carried out in triplicate using a biological class II safety cabinet. No change in color or turgidity was observed with the agent tested, proving the sterility of the product and that there was no trace of contamination. Forty larvae (in three replications) in the periods of 12, 24, and 48 h after sterilization, when transferred to diet, produced larvae, pupae, and total viability similar to the control (larvae without sterilization). However, for the 72- h treatment, larvae and total viability were significantly lower than for the other treatments. There was no significant difference for the pupal stage. The product tested was shown to be efficacious for use as a sterilizer of C. putoria eggs for all the parameters assessed.
Maggot debridement therapy (MDT) is a simple wound debridement technique. It is a natural treatment licensed by the Food and Drug Administration (FDA) and is increasingly used in the United States and in Europe. This treatment is safe when the larvae originate from laboratory stocks of eggs that have been sterilized. In this study, a simple, inexpensive microbe decontamination technique is described. It yields eggs that are free of chemical residues and are easy to handle, meeting the growing demand for medicinal larvae in hospitals or medical centers. Three treatments (T1, T2, T3) involving 3, 6, and 12 min of exposure to ultraviolet (UV-C) rays, respectively, were compared. Egg sterility was evaluated by culture in thioglycollate broth, incubated at 32°C ± 2.5°C under aerobic conditions for up to 14 d. The UV-C radiation sterilization process obtained satisfactory results after 12 min exposure (treatment 3). Larval viability was 57%, pupal viability was 54%, and 54% of the adults emerged. The sex ratio was 50%, within the expected values. There were no morphological abnormalities associated to the UV-C treatment in the flies. In conclusion sterilization by UV-C rays is indicated to obtain sterile larvae destined for MDT.
Antibiotic‐resistant bacteria pose a major threat to global health in the 21st century, requiring a quick, cheap and effective response from public health officials. This study evaluated the antimicrobial activity of native excretions/secretions (NES) produced by third instar (3 days old) larvae of Calliphora vicina using a protocol adapted from the Institute of Clinical and Laboratory Standards (CLSI). The microorganisms tested were: Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus and the fungus Candida albicans. After the incubation period, the suspensions were diluted and spread on nutrient agar plates to count the colony‐forming units. A turbidimetric test also was carried out to test the action of the NES of C. vicina against S. aureus, a very common bacterial species, with an enormous capacity for adaption and resistance, being one of the bacteria of medical importance that causes the most hospital and community infections in the world. According to our results, the NES of C. vicina exhibits antimicrobial activity at different dilutions, being most effective against the gram‐negative bacteria E. coli and K. pneumoniae.
We evaluate the effects the antibiotic Gentamicin on the development of Chrysomya putoria (Wiedemann, 1818). Third-generation, first-instar larvae were reared in a climatic chamber on 60 g of homogenate + agar 65% and were treated with three concentrations of Gentamicin: 4.44 mg/ml, 13.33 mg/ml, and 66.66 mg/ml. The control consisted of distilled water. The relationships between mean body mass of mature larvae (measured after diet abandonment, in batches of five individuals), duration of larval and pupal stages, and overall duration of development were analyzed. The actual sex ratio was compared against the expected using the chi square. None of the parameters measured differed significantly among the four treatments, with one exception: when Gentamicin concentration was 13.33 mg/ml, larval viability differed significantly from the control. All larvae from all treatments were considered normal. We conclude that the antibiotic did not significantly alter the development of C. putoria (Wiedemann) (Diptera: Calliphoridae).
Chrysomya putoria (Wiedemann) (Diptera: Calliphoridae), an Old World screwworm fly, is a species with potential for maggot therapy practice and has been described in myiasis and forensic entomology studies. The objective of the present study was to assess the action of different ciprofloxacin concentrations on the growth and development of C. putoria . First instar maggots of the third generation were raised on 60 g of chicken gizzard homogenate in 65% agar diet and received ciprofloxacin chloridrate. Each concentration of the antibiotic tested (3.33 µg/mL, 6.66 µg/mL, and 13.33 µg/mL) and the control (no antibiotic) were replicated four times (40 maggots/replication). The control received distilled water instead of the antibiotic. Maggots were kept in an acclimatized chamber at 30° C during the day and 28° C at night, with 70 + 10% RH and a 14:10 L:D photoperiod. They were weighed in batches of five and stored in test tubes sealed with nylon fabric and elastic. Microsoft Excel and STAT were used for the analysis. The variation among the maggot weight means and the duration of the maggot stage, pupal stage, and time to total development (neolarvae to adult) were analyzed by Student’s t -test (α= 5%). The viabilities and the normality rates were compared using ANOVA, and the expected sex ratio frequency was tested by the chisquared test (χ 2 ). There was no significant difference among the four treatments regarding mean individual maggot weight, mean duration of the maggot inoculation until abandonment, the duration of the maggot and pupal stages, and the total duration of all stages. The sex ratios found in the four treatments did not differ from the expected. Only treatment 2 (6.66 µg/mL concentration of ciprofloxacin) differed significantly from the control in maggot and total viability. The antibiotic did not seem to alter C. putoria development in the postembryonic period.
Here we evaluate the effects of different concentrations of the antibiotic ampicillin on the growth and development of Chrysomya putoria. Third-generation, fi rst instar larvae (L1) reared on 60 grams of homogenate+agar 65% were treated with ampicillin sodium. The experiment consisted of four replicates (40 larvae/replicate) of each antibiotic concentration tested (T1: 466μg/mL ; T2: 81.33 mg/mL and T3: 166.66mg/mL) and a T4: control. The body mass of the mature larvae, after they abandoned the diet, were recorded in batches of fi ve. The variation between the mean body mass of larvae and the duration of larval and pupal stages, and overall duration of the development, viability and normal rates were analyzed by ANOVA. There were no signifi cant differences between the four treatments in the following parameters: body mass of larvae that discontinued the diet as well as the duration of larval, pupal, and total development. The sex ratios found in the four treatments did not differ from those expected. Normality rates were 100% for all treatments. There were no signifi cant differences between treatments for larval and overall viability, but pupal viability differed signifi cantly between T1 and the control, T1 and T2, and between the control and T3. The antibiotic did not appear to signifi cantly alter the development of C. putoria.
Chicken gizzard was tested as an alternative to beef diet for creating Chrysomya megacephala (Fabricius) to reduce costs and optimize the time of diet preparation and maintenance of the colony diet. Forty newly hatched larvae were placed in 100 grams of diet: gizzard and meat (control). The experiment was performed in triplicate. The mass of mature larvae was recorded in semi-analytical balance and separate batch of five. After emergence, three replicates of 10 pairs of adults were formed. The post-embryonic development was evaluated by Tukey’s test at a 5% level of significance and longevity was adjusted Weibull regression. The mass of mature larvae showed no significant difference. The larval stage of flies reared in meat was longer. Pupae and newly emerged adults showed no significant differences as a function of diet. The increased mortality rate after after 22 days in both experimental diets. Greater longevity was achieved with diet gizzard, which was more efficient than the meat on the biological parameters studied. Estudo Comparativo entre a Moela de Frango e Carne Bovina como Dietas e suas Influências sobre o Desenvolvimento Pós-Embrionário e Longevidade de Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae) Resumo. Moela de Frango foi testada como dieta alternativa à carne para criação de Chrysomya megacephala (Fabricius) visando reduzir custos e otimizar o tempo de preparação da dieta e manutenção da colônia. Quarenta larvas recém eclodidas foram colocadas em 100 gramas de dieta: moela e carne (controle). O experimento foi realizado em triplicata. A massa das lavas maduras foi registrada em balança semi-analítica e separada em lotes de cinco. Após a emergência, foram formadas três repetições de 10 pares de insetos adultos. O Desenvolvimento pós-embrionário foi avaliado pelo teste Tukey com nível a 5% de significância e a longevidade foi ajustada por regressão Weibull. A massa das lavas maduras não apresentou diferença significativa. A fase larval de moscas criadas em carne foi mais longa. Pupa e adultos recém-emergidos não apresentaram diferenças significativas em função da dieta. A mortalidade aumentou após 22 dias de experimento em ambas as dietas. Uma maior longevidade foi conseguida com a dieta moela, que se mostrou mais eficiente do que a carne sobre os parâmetros biológicos estudados.
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