Samples of sewage from a university hospital and a chemistry technical school were analysed for the percentage of bacterial tolerance to chromium (Cr), silver (Ag) and mercury (Hg). Additionally, we investigated the effect of these metals on pigmentation and on some enzymatic activities of the metal tolerant strains isolated, as well as antimicrobial resistance in some metal tolerant Enterobacteriaceae strains. Tolerance to Cr was observed mainly in Gram positive bacteria while in the case of Ag and Hg the tolerant bacteria were predominately Gram negative. Hg was the metal for which the percentage of tolerance was significantly higher, especially in samples from the hospital sewage (4.1%). Mercury also had the most discernible effect on color of the colonies. Considering the effect of metals on the respiratory enzymes, one strain of Ag-tolerant Bacillus sp. and one of Hg-tolerant P. aeruginosa were unable to produce oxidase in the presence of Ag and Hg, respectively, while the expression of gelatinase was largely inhibited in various Gram negative strains (66% by Cr). Drug resistance in Hg-tolerant Enterobacteriaceae strains isolated from the university hospital sewage was greater than 80%, with prevalence of multiple resistance, while the Agtolerant strains from the same source showed about 34% of resistance, with the predominance of monoresistance. Our results showed that, despite the ability of metal tolerant strains to survive and grow in the presence of these elements, the interactions with these metals may result in metabolic or phisiological changes in this group of bacteria.

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.

Staphylococcus aureus is a pathogen commonly resistant to antibiotics. Biofilm formation is one of the important factors related to its virulence. Non-antibiotics drugs, such as nonsteroidal anti-inflammatory agents (NSAIDs), have been studied as an alternative for treating infections by multiresistant pathogens and biofilm-associated infections. In this study, the effects of NSAID sodium diclofenac on growth inhibition and biofilm formation of S. aureus were evaluated. The minimum inhibitory concentration (MIC) of diclofenac for fifty isolates ranged from 200 to 400 μg/mL. Diclofenac sub-MICs induced biofilm in 32.3% of biofilm-negative strains in tryptic soy broth. All biofilms induced by the drug showed a PIA- (polysaccharide intercellular adhesion-) independent composition, and the scanning electron microscopy showed that the induced biofilm presented a very discrete matrix. The combination of diclofenac with rifampicin sub-MICs induced strong production of PIA-dependent biofilm in three of four strains, while combination of NSAID with NaCl induced the formation of partially polysaccharide biofilm in two strains and PIA-independent biofilm in another strain. The combination of NSAID with glucose resulted in PIA-independent biofilms in all four strains tested. The results showed that diclofenac can commonly induce biofilm production by a PIA-independent pathway. However, when this NSAID is combined with other types of inducing agents, the composition of the biofilm produced may vary.

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.