Prevalence of bovine mastitis-related pathogens, identified by mass spectrometry in flies (Insecta, Diptera) captured in the milking environment.

Diptera (Insecta) are able to transmit approximately 200 pathogenic microorganisms to humans and animals, causing more than 65 diseases, including bovine mastitis, which constitutes a major cause of economic losses in the dairy industry. In this scenario, 217 adult specimens of Diptera were collected from nine farms and identified to the family and/or species level. Of the 11 families recorded, Muscidae was the most prevalent (152 out of 217; 70%). All Diptera specimens were subjected to microbiological culture using conventional and selective media, and isolates were then identified at the species level by mass spectrometry. In total, 275 microorganisms were identified, with a predominance of pathogens related to environmental bovine mastitis (166/275 = 60·4%), that is, Enterococcus species (70/275 = 25·4%) and Escherichia coli (49/275 = 17·8%). Nontraditional agents related to bovine mastitis (called miscellaneous) were detected in 28% (77/275), as well as microorganisms with well-known zoonotic behaviour (e.g. Bacillus cereus). This is study contributes with knowledge of diversity of microorganisms carried by Diptera in the dairy environment, including pathogens associated with environmental and contagious bovine mastitis, and agents with human relevance. To our knowledge, a three-part chromogenic selective medium used to microbial culture of milk on farms was used for the first time to identification of pathogens in Diptera.

Egg Developmental Time and Survival of Chrysomya megacephala and Chrysomya putoria (Diptera: Calliphoridae) Under Different Temperatures.

Chrysomya megacephala (F.) and Chrysomya putoria (Wiedemann) (Diptera: Calliphoridae) are considered of forensic, medical, and veterinary importance in Brazil because of their necrophagous and synanthropic behaviour. The development of flies can be influenced by temperature, and species from the same genus usually have different responses to external variables. The egg development of blow fly can be a useful complementary technique to estimate the minimum postmortem interval. Thus, this study aimed to compare the egg developmental time and survival of C. megacephala and C. putoria at different temperatures to determine the optimal temperature for egg development and the linear regression for developmental time and temperature, thereby determining the minimum threshold (t) and thermal summation constant (K) for each species. Adults of both species were collected in the region of Campinas city, São Paulo state, Brazil. Eggs were incubated at eight constant temperatures between 05 ± 1°C and 35 ± 1°C and the egg developmental time and survival were evaluated. There was no egg survival at 5 and 10°C. The K for C. megacephala and C. putoria were 179.41 HD and 189.94 HD, respectively. The regression slopes and t (10°C) were similar for both species. The optimal temperature for egg survival was between 25 and 35°C, for C. megacephala and 20 and 30°C, for C. putoria. The present data were similar to most data available in the literature, but differences in the same species are a possibility.

Effect of nandrolone decanoate on the development of three species of Chrysomya (Diptera:Calliphoridae), flies of forensic importance in Brazil.

Necrophagous insects are valuable tools for postmortem interval (PMI) estimation or for determining the cause of death. Due to the increase in deaths related to drug abuse, it is crucial to know how these substances affect the development of flies that feed on corpses, to avoid errors in the PMI estimates. This study evaluated the effect of nandrolone decanoate, an anabolic androgenic steroid, on the development ofimmatures of Chrysomya megacephala (F.), Chrysomya putoria (Wiedemann), and Chrysomya albiceps (Wiedemann) (Diptera:Calliphoridae) when added to an artificial rearing diet. Four experimental groups were delineated: three of them were given diets containing 4.5, 22.5, or 45 mg/kg nandrolone decanoate; and a drug-free control group. Weights were recorded at 12-h intervals from larval eclosion to pupation. No statistically significant differences were observed in mean larval weights, emergence interval, or emergence rates for all groups. However, differences in the three species were observed during the larval development. Initially, C. putoria reared in the highest concentration of decanoate showed greater weight gain. However, at older ages, immatures reached lower mean weights than the control group. For C. albiceps, the highest concentration of decanoate contributed to an effective lack of weight gain during almost the entire course of larval development. Therefore, the influence of any drug on development should always be considered.

Breeding of Ornidia obesa (Diptera: Syrphidae: Eristalinae) on pig carcasses in Brazil.

Ornidia obesa F. (Diptera: Syrphidae) is usually neglected in forensic entomology, although adults are rather frequent on vertebrate carrion. In this study, conducted in southeastern Brazil in 2008, we used two pig carcasses, one killed by cocaine overdose and the other by shooting, to evaluate mainly the possible influences of the type of death on the larval development of O. obesa in the pig remains. We recorded the breeding of 218 adult specimens of this syrphid fly from the carcass killed by shooting, and none from the carcass killed by cocaine. These observations may open a new perspective for the use of O. obesa in forensic studies, considering its breeding preferences and its complete development on vertebrate carrion.

Phasmatodea (Arthropoda, Insecta) in Brazil: Status, New Record, and Proposal for Using Molecular Tools to Assist in Species Identification.

Phasmatodea (Arthropoda, Insecta) comprises phytophagous insects, popularly known as stick bugs because most of them resemble dry branches. Currently, more than 3000 species are known, the majority of which can be found in tropical regions of the world. In Brazil, it is estimated that there are approximately 200 species, but these numbers may not truly reflect the group's richness. A high degree of intraspecific polymorphism, a low number of specimens sampled, a shortage of specialized literature, and having only a small number of experts have amplified the difficulties in studying this order. Therefore, this study aims to present a historical survey of the Phasmatodea studies that have been conducted in Brazil thus far, indicating gaps in knowledge and discussing perspectives to expand understanding within this group. Furthermore, to the best our knowledge, the molecular characterization of the cytochrome oxidase subunit I gene of mitochondrial DNA of two species of phasmids found in Brazil, Cladomorphus phyllinus Gray, 1835, and Pseudophasma missionum Piza, 1981, is presented for the first time. In addition, this study records for the first time the occurrence of P. missionum in Brazilian territory.

First Report on the Use of Larvae of Cochliomyia macellaria (Diptera: Calliphoridae) for Wound Treatment in Veterinary Practice.

Factors contributing to the delay or prevention of the natural wound healing process include infection and ineffective conventional treatment. Alternative therapies, such as the maggot debridement therapy (MDT), may be helpful for successful treatment in these cases. Aiming to disseminate information about the possibility of using other species of flies for wound treatment, besides the best known Lucilia sericata Meigen, 1826 (Diptera, Calliphoridae), we report here a successful MDT case with the application of Cochliomyia macellaria (Fabricius, 1775) larvae to treat an infected wound with extensive area of necrotic tissue in a dog. Five sterile larvae were applied to each square-cm of lesion and kept on the animal for only 48 h. The healing was successful, from both qualitative and quantitative points of view. The ratio of wound healing (RWH) reached almost 50% on the 5th day and 100% on the 14th day after MDT. Although the overall animal prognosis had been unfavorable, mainly due to the sepsis, the patient began to recover and had improved clinical condition from the fifth day after MDT. This study shows the importance and effectiveness of MDT in promoting faster and more complete healing of a complex wound.

Optimized Pitfall Trap Design for Collecting Terrestrial Insects (Arthropoda: Insecta) in Biodiversity Studies.

Pitfall traps are commonly used for the collection of terrestrial insects in ecology and biology studies; they are relatively straightforward to manufacture and there is a large variety of models described in the literature. However, they present a few drawbacks: (i) the removal and transport of the collected material are not practical; (ii) they have low resistance and durability; (iii) they fail to correctly protect the attractive bait against adverse weather conditions and scavengers, and (iv) evaporation of the liquid used inside the trap. We proposed an optimized pitfall trap design for terrestrial insect collection made from cheap and easily accessible materials. The new design allows the transfer of the collected material to the lab by removing only that part of the trap where the insects have been captured; the other part remains in its original place. Thus, the proposed trap allows easier operation since there is no need to transport water to replenish the traps after each transfer; in addition, there is less volume and weight to be carried. The trap can remain in the field for months because of the durability of its material. Furthermore, the collected material is better protected against adverse weather conditions and scavengers. Currently, an efficient and rapid sampling strategy in the field is of global interest to understand mechanisms that can contribute to the monitor changes in phenology, succession, and biodiversity.

First Record of Larval Secretions of Cochliomyia macellaria (Fabricius, 1775) (Diptera: Calliphoridae) Inhibiting the Growth of Staphylococcus aureus and Pseudomonas aeruginosa.

Maggot debridement therapy (MDT) consists on the intentional and controlled application of sterilized larvae of the order Diptera on necrotic skin lesions with the purpose of cleaning necrotic tissue and removing pathogenic bacteria. During MDT, a marked antimicrobial activity has been reported in literature specially associated with antibacterial substances from Lucilia sericata (Meigen); however, regarding Cochliomyia macellaria (Fabricius), little is known. This study aimed to evaluate in vitro inhibition of bacterial growth of Pseudomonas aeruginosa and Staphylococcus aureus in contact with excretions and secretions (ES) from C. macellaria larvae. Larval ES were extracted in sterile distilled water and divided in three groups: ES, containing 400 µL of autoclaved ES; ES+BAC, containing 400 µL of autoclaved ES+0.5-µL bacterial inoculum; and CONT-BAC, containing 400 µL of sterile distilled water +0.5 µL of bacterial inoculum. Aliquots of each experimental group were plated by spreading onto Petri dishes. Seedings were made at 0, 1, 2, 4, and 12 h after the extraction of ES. In ES+BAC groups, inhibition of S. aureus was verified between times 1 and 2 h and P. aeruginosa was inhibited between 0 and 4 h. There was no growth observed in any ES group. In the CONT-BAC groups, the number of colonies from time 4 h became countless for S. aureus and decreased for P. aeruginosa. As reported in the literature, we note here that ES have excellent bactericidal activity for both gram-positive and gram-negative bacteria, and this study shows for the first time the action of the bactericidal activity of exosecretions of C. macellaria against S. aureus and P. aeruginosa.

Eimeria curvata n. sp.(Apicomplexa: Eimeriidae) in Columbina talpacoti and Scardafella squammata (Aves: Columbidae) from Brazil.

Eimeria curvata is a new coccidian described in the doves Columbina talpacoti and Scardafella squammata from western of the State of São Paulo, Brazil. The oocysts are ovoid to ellipsoid, 18.3 (17-19) microm x 15.5 (15-17) microm, with a shape index of 1.2 (1.1-1.3). The wall is colorless, smooth and double-layered. A polar granule is present, but there is no micropyle or oocyst residuum. The sporocysts are elongate, 12.3 (11.5-13) microm x 5.8 (5.5-6) microm with a curved anterior portion and a smooth, thin, single-layered wall. The Stieda body is protuberant and nipple-like; there is no substieda body. The sporozoites lie head-to-tail in the sporocyst and contain a large refractile body at the extremities. The sporocyst residuum contains small granules uniformly distributed in the middle of the sporocyst. The prevalence of E. curvata n. sp. was 17.4% and 12.8% in C. talpacoti and S. squammata, respectively.

A checklist of arthropods associated with pig carrion and human corpses in southeastern brazil.

Necrophagous insects, mainly Diptera and Coleoptera, are attracted to specific stages of carcass decomposition, in a process of faunistic succession. They are very important in estimating the postmortem interval, the time interval between the death and the discovery of the body. In studies done with pig carcasses exposed to natural conditions in an urban forest (Santa Genebra Reservation), located in Campinas, State of São Paulo, southeastern Brazil, 4 out of 36 families of insects collected - Calliphoridae, Sarcophagidae, Muscidae (Diptera) and Dermestidae (Coleoptera) - were considered of forensic importance, because several species were collected in large numbers both visiting and breeding in pig carcasses. Several species were also observed and collected on human corpses at the Institute of Legal Medicine. The species belonged to 17 different families, 6 being of forensic importance because they were reared from human corpses or pig carcasses: Calliphoridae, Sarcophagidae, Muscidae, Piophilidae (Diptera), Dermestidae, Silphidae and Cleridae (Coleoptera). The most important species were: Diptera - Chrysomya albiceps, Chrysomya putoria, Hemilucilia segmentaria, Hemilucilia semidiaphana (Calliphoridae), Pattonella intermutans (Sarcophagidae), Ophyra chalcogaster (Muscidae), Piophila casei (Piophilidae); Coleoptera - Dermestes maculatus (Dermestidae), Oxyletrum disciolle (Silphidae) and Necrobia rufipes (Cleridae).

Miíase primária em coelho doméstico causada por Lucilia eximia ( Diptera: Calliphoridae ) no Brasil: relato de caso / Primary myiasis in a domestic rabbit caused by Lucilia eximia ( Diptera: Calliphoridae ) in Brazil: case report

Relata-se a ocorrência de miíase primária em coelho doméstico Oryctolagus cuniculus (Lagomorpha: Leporidae) causada por Lucilia eximia (Diptera: Calliphoridae) em área urbana do município de Campinas, São Paulo.

The occurrence of primary myiasis in a domestic rabbit Oryctolagus cuniculus (Lagomorpha: Leporidae) caused by Lucilia eximia (Diptera: Calliphoridae) is reported in an urban area in the city of Campinas, São Paulo, Brazil.

A checklist of arthropods associated with pig carrion and human corpses in Southeastern Brazil

Necrophagous insects, mainly Diptera and Coleoptera, are attracted to specific stages of carcass decomposition, in a process of faunistic succession. They are very important in estimating the postmortem interval, the time interval between the death and the discovery of the body. In studies done with pig carcasses exposed to natural conditions in an urban forest (Santa Genebra Reservation), located in Campinas, State of São Paulo, southeastern Brazil, 4 out of 36 families of insects collected - Calliphoridae, Sarcophagidae, Muscidae (Diptera) and Dermestidae (Coleoptera) - were considered of forensic importance, because several species were collected in large numbers both visiting and breeding in pig carcasses. Several species were also observed and collected on human corpses at the Institute of Legal Medicine. The species belonged to 17 different families, 6 being of forensic importance because they were reared from human corpses or pig carcasses: Calliphoridae, Sarcophagidae, Muscidae, Piophilidae (Diptera), Dermestidae, Silphidae and Cleridae (Coleoptera). The most important species were: Diptera - Chrysomya albiceps, Chrysomya putoria, Hemilucilia segmentaria, Hemilucilia semidiaphana (Calliphoridae), Pattonella intermutans (Sarcophagidae), Ophyra chalcogaster (Muscidae), Piophila casei (Piophilidae); Coleoptera - Dermestes maculatus (Dermestidae), Oxyletrum disciolle (Silphidae) and Necrobia rufipes (Cleridae).