Fly Colonies Research Articles

Examining imidacloprid behaviorally resistant house flies (Musca domestica L.) (Diptera: Muscidae) for neonicotinoid cross-resistance.

The house fly (Musca domestica L.) is a ubiquitous fly species commonly associated with confined animal and urban waste storage facilities. It is known for its pestiferous nature and ability to mechanically vector numerous disease-causing pathogens. Effective control of adult house fly populations has traditionally relied upon insecticidal food baits; however, due to the overuse of insecticides, resistance has proven to yield many insecticidal baits and chemical classes less effective. Imidacloprid, the most widely used neonicotinoid, has been formulated and commonly used in house fly baits for over 2 decades. However, widespread evidence of physiological and behavioral resistance to imidacloprid has been documented. While previous studies have investigated the mechanisms of behavioral resistance to imidacloprid in the house fly, it remains unclear whether behavioral resistance is specific to imidacloprid or if behavioral cross-resistance exists to other compounds within the neonicotinoid class of insecticides. The current study used no-choice and choice-feeding bioassays to examine a lab-selected imidacloprid behaviorally resistant house fly colony for cross-resistance to other insecticides in the neonicotinoid chemical class. All flies exhibited high mortality (97-100%) in no-choice assays, even when exposed to imidacloprid, indicating physiological susceptibility to all tested neonicotinoids. House flies exhibited high mortality (98-100%) in choice assays when exposed to all neonicotinoid insecticides tested besides imidacloprid. These results confirm that imidacloprid behavioral resistance is specific to the compound imidacloprid and that alternative neonicotinoids remain viable options for control. Our study showed no evidence of behavioral cross-resistance to other compounds in the neonicotinoid class.

Assessment of optimal blood exposure time at 37 °C during in vitro tsetse flies feeding for quality production in mass-rearing colonies.

Control of African animal trypanosomosis is implemented through an integrated control strategy, with the sterile insect technique (SIT) as one of its components. The SIT requires mass rearing of tsetse fly colonies using an in vitro feeding system. The exposure of blood at 37 °C on heating plates over time can have an impact on the quality of fly productivity. In this study, we investigated the survival and fecundity of adult tsetse flies fed at 37 °C on 8 blood exposure times ranging from 30 min to 4 h with increments of 30 min (treatment 1, flies were fed 30 min after exposure to blood at 37 °C; treatment 2, 1 h and so on until treatment 8 [4 h after]) in order to determine the optimal exposure time. In addition, bacterial growth in blood from each treatment was assessed by agar culture at 37 °C for 72 h. The results showed that the adult female survival rates were similar regardless of the treatment. For males, only those of treatment 1 (30 min) showed a marginal lower survival than those of treatments 7 and 8 fed after 3 h 30 min and 4 h of blood exposure, respectively. Over the 4-h interval of blood exposure at 37 °C, the results showed that the number of pupae produced per initial female and pupal weight tended to increase with exposure time, but the differences were not significant. We discuss the implications of these results on tsetse mass rearing for the SIT program.

Multi-objective Scheduling of Tasks in Cloud Computing Using Fire Fly and Bee Colony Algorithms

The problem of allocating hardware and processing resources in cloud computing is considered in this research. Here, a virtual bee is assigned to any task that dynamically seeks the most appropriate computing resources. Each available computing source also has a virtual fire fly that its brightness is proportional to the amount of processing unit and the amount of computing residue that examines the computing source. The problem of distribution of computing resources among existing tasks will be investigated using the greedy technique of calculating the location of each task in general. The current method has many advantages compared to the random distribution method, but to complement the other topics, researchers can offer other tactics of distribution of computing resources simultaneously with the method proposed in this study and compare their performance with the implementation of both simulations. The simulation results show that the performance criteria of the cloud computing system such as time and computing resources have been greatly improved.

Validation study of Yunnan ethnic culture industry stock model under epidemic situation based on chaotic particle swarm optimization neural networks

The new crown pneumonia epidemic is raging, in the context of global integration, the scope of the impact of this sudden event spread around the world, the stock market has not been spared, the financial risk has increased dramatically compared with the past, the emergence of the epidemic has led to the spread of investor panic, March 2020, the U.S. S&P 500 index appeared in the four plunge, and led to the market trading meltdown, the world’s financial markets have had an extremely serious impact. The study of the impact of Xin Guan Pneumonia on the company’s stock returns is not only conducive to enriching the theoretical study of public health emergencies, but also conducive to improving the coping strategy, stabilizing the general economic market, and enhancing the public’s awareness of risk response. This paper compares the effect of the four intelligent algorithms of chaotic particle swarm algorithm, chaotic bee colony algorithm, chaotic fruit fly algorithm and chaotic ant colony algorithm combined with neural network on the prediction of the stock price trend of Yunnan national culture, and the study shows that the speed of convergence of the chaotic particle swarm optimization neural network and the speed of descent is better than that of the two models of chaotic fruit fly and chaotic bee colony, and the coefficients of decision of the chaotic particle swarm optimization neural network are higher than that of the other three models, and the errors are lower than the other three models. Indexes are lower than the other three models and have high accuracy in stock prediction of Yunnan ethnic culture, this finding emphasizes the potential of PSO-BP model to provide robust stock market prediction, which is important for both investors and policy makers in dealing with volatile market conditions.

Open and Closed Black Soldier Fly Systems Tradeoff Analysis

When it comes to waste management, it is essential to consider human and environmental safety, financial feasibility, and social benefits. So often, one of these factors takes priority over the others. However, integrated social, environmental, and economic benefits are more apparent with insect-based waste treatment technology like the Black Soldier Fly (BSF) system. BSF waste treatment is an innovative and adaptable technique that offers sustainable benefits to communities in developing countries because it can be designed to be as simple or complex as required. Depending on the local context, simple (open) systems based on naturally occurring flies or more complex (closed) systems based on captured fly colonies are viable. However, what are the environmental tradeoffs when choosing between these two systems? The surge in the use of both BSF systems makes this consideration imperative. Furthermore, until now, the environmental and social impacts of open BSF waste treatment systems have not been compared. The environmental tradeoffs in implementing an affordable, socially accepted, open BSF biowaste treatment system are explored in this study to address this gap. Co-production for delivering public services was used to develop an accepted BSF system through a case study by applying qualitative interaction research methodologies. A Life Cycle Assessment (LCA) was then used to compare the environmental impacts of natural ovipositing BSF systems compared to captured BSF systems. Natural ovipositing BSF (open) systems, in comparison, have reduced climate change (33%) and water, land, and energy use (55%, 37% and 32%) while also motivating the development of socially accepted infrastructure measured through community engagement. This mixed-methods approach facilitates the development of contextually appropriate technology in low-income communities in developing and developed countries while also reducing environmental consequences.

Toward optimising reproductive output of Eristalis tenax (Diptera: Syrphidae) for commercial mass rearing systems

AbstractDeveloping mass rearing systems for the drone fly, Eristalis tenax, is a crucial step toward its use as a complementary commercial pollinator. To meet the timing of commercial needs for E. tenax, there is significant value to both predicting and managing reproductive output and development within the rearing protocol. To help achieve this, our study focuses on the laboratory manipulation of adult mating and the timing of the development of eggs in E. tenax. To stagger colony cohorts, egg rearing temperatures ranging from 12 to 30°C were found to be suitable for both successful egg development and hatching viability (97 to 28.3 h to first hatch, respectively). The mating window for E. tenax females was established to commence from 2 weeks post eclosion and reached 75 ± 11% mated at 7 weeks. Reproductive output over 12 weeks was assessed in separate cage manipulation trials: (1) varying the sex ratio with 20:40, 30:30 and 40:20 female and male flies respectively per cage and (2) varying the adult density per cage with 15:15, 30:30, 60:60 and 120:120 female and male flies per cage. Female percentage mated and egg cluster size, which averaged 200.6 ± 4.3 eggs per cluster, did not change between treatments in the sex ratio and density trials. Egg cluster output per female was significantly reduced for treatments across both trials, which had more than 30 females in a cage. A stocking rate of 15:15 produced 86% more eggs per female than expected, a percentage well above that of all other treatments. However, the highest stocking density produced the most eggs when assessed at a per cage level. Fly survival was significantly different between the sexes across both trials with males dying earlier in cages stocked with more females than males. Although the fly colonies were held at constant temperatures and light conditions for 6 months, we found evidence of endogenous overwintering behaviours among flies resulting in lower mating rates, egg hatching success and greater longevity among flies studied over winter.

Gene expression differentials driven by mass rearing and artificial selection in black soldier fly colonies.

The micro-evolutionary forces that shape genetic diversity during domestication have been assessed in many plant and animal systems. However, the impact of these processes on gene expression, and consequent functional adaptation to artificial environments, remains under-investigated. In this study, whole-transcriptome dynamics associated with the early stages of domestication of the black soldier fly (BSF), Hermetia illucens, were assessed. Differential gene expression (DGE) was evaluated in relation to (i) generational time within the cultured environment (F2 vs. F3), and (ii) two selection strategies [no artificial selective pressure (NS); and selection for greater larval mass (SEL)]. RNA-seq was conducted on 5th instar BSF larvae (n=36), representing equal proportions of the NS (F2=9; F3=9) and SEL (F2=9; F3=9) groups. A multidimensional scaling plot revealed greater gene expression variability within the NS and F2 subgroups, while the SEL group clustered separately with lower levels of variation. Comparisons between generations revealed 898 differentially expressed genes (DEGs; FDR-corrected p < 0.05), while between selection strategies, 213 DEGs were observed (FDR-corrected p < 0.05). Enrichment analyses revealed that metabolic, developmental, and defence response processes were over-expressed in the comparison between F2 and F3 larvae, while metabolic processes were the main differentiating factor between NS and SEL lines. This illustrates the functional adaptations that occur in BSF colonies across generations due to mass rearing; as well as highlighting genic dynamics associated with artificial selection for production traits that might inform future selective breeding strategies.

High Temperature Limits of Survival and Oviposition of Phormia regina (Meigen) and Lucilia sericata (Meigen).

The temperature dependent development rates of blow flies allow blow flies to be used as biological clocks in forensic death investigations. However, the upper thermal limits of adult survival and oviposition, both required for producing larvae, remains largely unknown. Therefore, in this study we examined the impact of a range of temperatures between 37 °C and 44 °C on the likelihood of survival and egg-laying behavior of two species of medicolegal forensic importance, Lucilia sericata (Meigen) and Phormia regina (Meigen) (Diptera: Calliphoridae). To quantify the upper temperature limits of survival, adult fly colonies were exposed to 37 °C, 41 °C, 42 °C, 43 °C, and 44 °C for 24 h. Similarly for oviposition trials, adults of both species were exposed to 40 °C, 42 °C, and 43 °C with P. regina oviposition also observed at 41 °C. Trials lasted for 24 h with oviposition substrate replenished at the 12 h mark. A yes/no determination on egg deposition was made, eggs were counted, and a yes/no determination was made on egg hatch. Survival did not differ by species (p = 0.096). Overall, survival decreased with increasing temperatures, with ~100% at 37 °C, ~50% at 41 °C, ~37% at 42 °C, ~15% at 43 °C and 0% at 44 °C. Lucilia sericata laid eggs capable of hatch up to 43 °C, while Phormia regina egg-hatch was observed up to 41 °C. These results indicate a greater thermal tolerance of adult survival than for egg deposition and successful egg hatch, which supports previous experiments indicating blow flies stop laying eggs at sub-lethal temperatures. Furthermore, these data indicate that adult blow flies may find remains at or near time of death but may delay egg deposition until temperatures drop below an acceptable threshold.

Aspects of the biology of &lt;em&gt;Phlebotomus argentipes&lt;/em&gt; (Diptera: Psychodidae) in Kurunegala district

The sand fly species Phlebotomus argentipes is reported to be the suspected primary vector of leishmaniasis in Sri Lanka transmitting Leishmania donovani which causes visceral leishmaniasis. Thus, studies about some aspects of the reproductive biology of P. argentipes and the performances of immature stages are highly important for adopting control measures. Accordingly, sand flies were captured from the wild using five CDC light traps set in pre-identified places in the Kurunegala district. Gravid and fully engorged females were sorted out and they were directly placed for oviposition in plaster of Paris lined pots. The unengorged ones were allowed to feed on a mouse and placed for oviposition in similar pots. The rate of oviposition, egg hatching, and adult emergence was determined by making daily observations. Moreover, the duration of eggs, each larval instar, pupae, and adult longevity were recorded. According to the observations the life cycle of P. argentipes lasted about 3 to 4 months. The eggs were laid 10-13 days after the blood meal, and they hatched after 12-16 days. Similarly, each second instar, third instar, fourth instar and pupae spent 12-16 days for the emergence. However, the pupal duration was 8-10 days. Furthermore, the rate of oviposition was 15.8 per female and egg hatching and adult emergence was 62.52% and 52.32% respectively. The longevity of the emerged adults was 8-15 days. The information presented on P. argentipes in this study is highly important to initiate and maintain a sand fly colony under laboratory conditions for the implementation of successful control measures.

A novel low‐cost effective trap to capture sarcosaprophagous Diptera alive

Carrion flies are trapped to study their succession pattern and record their diversity or establish laboratory colonies for various purposes. They are typically collected manually, which is not only challenging and time-consuming, but may also result in incomplete or biased inventories. Passive trapping techniques commonly used damage the specimens and make them difficult to identify. We describe the construction of a trap to capture sarcosaprophagous Diptera alive. We tested six traps using six rabbit carcasses (Oryctolagus cuniculus L.) per trial over 1 year and recorded the diversity and succession of insects. We also established fly colonies of Lucilia sericata (Meigen) adults using captured individuals. A total of 27,428 insects (i.e., 8669 specimens in spring, 7969 specimens in summer, 8417 specimens in autumn and 2373 specimens in winter) belonging to 21 families and 43 species were collected over a period of 23, 19, 24 and 30 days in spring, summer, autumn and winter, respectively. The catch consisted mainly of the order Diptera (98.55%), but also Hymenoptera (1.45%). We successfully started a fly colony with 107 individuals in spring and added wild-type adults every season. The colonies were maintained for 1 year, during which the lifespan of the adult flies varied between 5 and 52 days. The device is easy to manufacture, low-cost, robust under field conditions, efficient and time-saving. It enables large samples to be collected within a short period of time over the entire trial period without affecting its condition or performance.

Characterization and Tissue Tropism of Newly Identified Iflavirus and Negeviruses in Glossina morsitans morsitans Tsetse Flies.

Tsetse flies cause major health and economic problems as they transmit trypanosomes causing sleeping sickness in humans (Human African Trypanosomosis, HAT) and nagana in animals (African Animal Trypanosomosis, AAT). A solution to control the spread of these flies and their associated diseases is the implementation of the Sterile Insect Technique (SIT). For successful application of SIT, it is important to establish and maintain healthy insect colonies and produce flies with competitive fitness. However, mass production of tsetse is threatened by covert virus infections, such as the Glossina pallidipes salivary gland hypertrophy virus (GpSGHV). This virus infection can switch from a covert asymptomatic to an overt symptomatic state and cause the collapse of an entire fly colony. Although the effects of GpSGHV infections can be mitigated, the presence of other covert viruses threaten tsetse mass production. Here we demonstrated the presence of two single-stranded RNA viruses isolated from Glossina morsitans morsitans originating from a colony at the Seibersdorf rearing facility. The genome organization and the phylogenetic analysis based on the RNA-dependent RNA polymerase (RdRp) revealed that the two viruses belong to the genera Iflavirus and Negevirus, respectively. The names proposed for the two viruses are Glossina morsitans morsitans iflavirus (GmmIV) and Glossina morsitans morsitans negevirus (GmmNegeV). The GmmIV genome is 9685 nucleotides long with a poly(A) tail and encodes a single polyprotein processed into structural and non-structural viral proteins. The GmmNegeV genome consists of 8140 nucleotides and contains two major overlapping open reading frames (ORF1 and ORF2). ORF1 encodes the largest protein which includes a methyltransferase domain, a ribosomal RNA methyltransferase domain, a helicase domain and a RdRp domain. In this study, a selective RT-qPCR assay to detect the presence of the negative RNA strand for both GmmIV and GmmNegeV viruses proved that both viruses replicate in G. m. morsitans. We analyzed the tissue tropism of these viruses in G. m. morsitans by RNA-FISH to decipher their mode of transmission. Our results demonstrate that both viruses can be found not only in the host’s brain and fat bodies but also in their reproductive organs, and in milk and salivary glands. These findings suggest a potential horizontal viral transmission during feeding and/or a vertically viral transmission from parent to offspring. Although the impact of GmmIV and GmmNegeV in tsetse rearing facilities is still unknown, none of the currently infected tsetse species show any signs of disease from these viruses.

Patterns of Genetic Diversity and Mating Systems in a Mass-Reared Black Soldier Fly Colony.

The black soldier fly (BSF), Hermetia illucens, is a promising candidate for the emerging insect farming industry with favourable characteristics for both bioremediation and production of animal delivered nutritive and industrial compounds. The genetic management of commercial colonies will become increasingly important for the sustainability of the industry. However, r-selected life history traits of insects pose challenges to conventional animal husbandry and breeding approaches. In this study, the long-term genetic effects of mass-rearing were evaluated as well as mating systems in the species to establish factors that might influence genetic diversity, and by implication fitness and productivity in commercial colonies. Population genetic parameters, based on microsatellite markers, were estimated and compared amongst two temporal wild sampling populations and four generations (F28, F48, F52, and F62) of a mass-reared colony. Furthermore, genetic relationships amongst mate pairs were evaluated and parentage analysis was performed to determine the oc-currence of preferential mate choice and multiple paternity. The mass-reared colony showed a reduction in genetic diversity and evidence for inbreeding with significant successive generational genetic differentiation from the wild progenitor population. Population-level analysis also gave the first tentative evidence of positive assortative mating and genetic polyandry in BSF. The homoge-neity of the mass-reared colony seems to result from a dual action caused by small effective popu-lation size and increased homozygosity due to positive assortative mating. However, the high ge-netic diversity in the wild and a polyandrous mating system might suggest the possible restoration of diversity in mass-reared colonies through augmentation with the wild population.

Efficient feature selection technique for brain tumor classification utilizing hybrid fruit fly based abc and ann algorithm

In medical science, information technology plays a main role in image processing for medicinal investigation and classification. For this reason “Magnetic Resonance Imaging” (MRI) can be useful today in the medical world which in turn is used to generate a comprehensive picture of the body's soft tissue and bones. The suggested technique utilizes an effective technique for brain tumor segmentation and classification. To segment and classify the brain tumor MR image by Modified fuzzy c means clustering (MFCM) and Artificial Neural Network (ANN). From the input image the proposed method extracts the shape, intensity and texture features. Here the extracted features are optimally selected by means of Hybrid Fruit fly and artificial bee colony (HFFABC). Both the segmentation and classification performance of the implemented technique is estimated in terms of “Accuracy, Sensitivity, Specificity, Ppv, Npv, Fpr And Fnr”. The implementation result shows the efficiency of proposed tumor segmentation and classification technique in segmenting the tissues perfectly from the MRI images of human nervous system. The classification performance of the implemented techniques is evaluated in terms of accuracy, sensitivity and specificity. The implementation results show the sensitivity, specificity & efficiency as 98.1%, 99.8% and 99.59%respectively.

Lucifensin, Chymotrypsin Proteins and Molecular Characterization of Lucilia sericata

Lucilia sericata, one of the most common species of the Calliphoridae family, is found in large numbers around droppings, garbage and carcasses. This fly species is important in medicine, forensics and veterinary medicine. The larvae of the parasite are important both in veterinary medicine and in combating of the animal diseases, as they cause significant losses in animal production. Since they are one of the first fly colonies to settle on corpses, they can also be used in determining the time of death in the field of forensic medicine. L.sericata larvae used in Maggot debridement treatment (MDT) which is a treatment method with fly larvae, help wound healing by destroying necrotic tissues and infectious agents in wounds. While the larvae protect themselves from polymicrobial flora with the proteins they secrete; at the same time, they make an interesting contribution to wound healing with these molecules secreted. One of the most important molecules discovered in recent years is lucimycin which has an antifungal effect. In addition, lucifensin and chymotrypsin secretions have gained importance in recent years due to their antibacterial effects and especially their effects on resistant gram-negative and positive bacteria. There is a need for the discovery of the molecules that can be alternative in the treatment of non-healing wounds or that can be applied together with existing antibiotics. It is necessary to investigate the antimicrobial characterization of the compounds involved in maggot therapy and their mechanisms. The aim of this study was to clone, molecular characterization and analysis of the antigenic structures of lucifensin and chymotrypsin genes, which are important defensin molecules secreted by L.sericata larvae used in MDT. Primarily, the cultivation of L.sericata colonies to be used in molecular studies were performed. Later, RNA isolation and cDNA synthesis from larvae were carried out. Lucifensin and chymotrypsin genes were individually inserted into the pJet1.2 plasmid by cloning reactions. The presence of the recombinant plasmid was confirmed by PCR screening and DNA sequence analysis methods in all steps. Nucleotide and amino acid based molecular characterizations of these two genes, which are important larval components in wound treatment, have been made. Antigenic regions and three-dimensional structures of the proteins were obtained. The isolate numbered MT495795 of the L.sericata lucifensin gene and the isolate numbered MT495794 of the chymotrypsin gene were registered to GenBank. This data reported for the first time in the Republic of Turkey will contribute to the literature. From the beginning of the 20th century until the discovery of the antibiotics, MDT was applied especially on soldiers but did not find much application area after the discovery of the antibiotics. Drug resistance, which is the most important problem encountered in the treatment of the wounds today, has led to the recall of MDT and its mechanism of action. In this study the data, obtained will constitute a source for the multidisciplinary studies of the scientists from different fields on the discovery and applicability of the important moleculesin the treatment of the wounds.

Interaction of age and temperature on heat shock protein expression, sperm count, and sperm viability of the adult black soldier fly (Diptera: Stratiomyidae)

Information regarding black soldier fly (Diptera: Stratiomyidae) adult biology is vital as this is the life stage that produces eggs and thus drives population size. The goal of this study was to determine key biological characteristics of adult black soldier flies as they age in relation to: (1) the thermal preferences (Tsel) of males and females; (2) the impact of temperature on heat shock protein expression in males and females; as well as (3) the sperm count; and (4) the sperm viability in males. Aging significantly impacted male and female temperature preferences. Young males (&lt;24-h-old) preferred warmer temperatures (median=24.3 °C, range=19.3-28.2 °C) compared to females of the same age (median=20.2 °C, range=15.4-26.2 °C). However, in older adults (i.e. 72-h-old males and 48-h-old females), temperature preferences converged between 21 and 24 °C. Temperatures tested did not impact hsp expression in males or females. However, aging males, but not females, had increased expression of the heat shock proteins (hsp) hsp70 and hsp90. Furthermore, age impacted sperm count but not sperm viability in males. In particular, 48-h-old males had the greatest sperm count (322.5/sample) and sperm viability (60-78%) compared to all other aged males. Thermal data in conjunction with sperm data potentially explain why early thermal segregation behaviour between males and females occurs. Once adult males and females reached 72-h-old and 48-h-old, respectively, they exhibited a common thermal preference, which coincided with the greatest number of viable sperm in males. Forcing adults into environments (i.e. cages) outside these selected preferences could result in premature or delayed mating or low fertilisation rates. Future research exploring cage design and conditions are needed to optimise black soldier fly colony maintenance and fertile egg production, and can leverage information such as the results described here.

Molecular Characterization of Lucimycin Gene of Lucilia sericata

Lucilia sericata, a member of the Calliphoridae family, is one of the most common species in the genus Lucilia. Medical importance of L.sericata stems from its use in maggot debridement therapy (MDT). MDT is the name of L.sericata larvae being sterilized and used in the treatment of non-healing wounds. L.sericata maggots used in the treatment of chronic and non-healing wounds (decubitus ulcer, venous leg ulcer, diabetic foot ulcer, etc.) clean the wounds with the help of secreted proteolytic trypsin and lucimycin -like enzymes. The aim of the study was to determine the molecular characterization of lucimycin gene obtained from L.sericata larvae in MDT by using molecular methods and to contribute to the literature. In this study, continuous production of adult colonies of L.sericata species was carried out in insectarium unit where conditions such as light, humidity and temperature were formed. The life cycle of L.sericata was followed and the production of eggs, larvae, pupae, adult flies and fly colonies of the species were formed. In the third stage larvae obtained from adult flies in the insectarium unit, RNA was isolated and subsequently cDNA synthesis was performed by reverse transcription. Polymerase chain reaction (PCR) analysis of the synthesized cDNAs with the specific primers designed for the lucimycin gene of L.sericata was performed and the obtained amplicons were cloned into pJET1.2/blunt vector and the plasmid was purified. The recombinant plasmids were sequenced with vector-specific primers and target gene region sequences were obtained. After the molecular characterization of the isolate with nucleotide sequences was determined, it was registered to GenBank database with the accession number MF964229. The PCR product of 288 bp was obtained from the cDNA obtained from the larvae of L.sericata produced in the insectarium unit by PCR using lucimycin specific primers. The PCR product imaged on the gel was purified by transformation and subsequent colonies were screened to see whether they contained recombinant plasmids. Three of the colonies were identified as recombinant plasmids containing L.sericata lucimycin gene by PCR screening. From three colonies confirmed by PCR screening, recombinant plasmids containing L.sericata lucimycin gene were purified by miniprep. The recombinant plasmid product was confirmed to contain the L.sericata lucimycin gene by PCR from a total of 20 μl of the recombinant plasmid miniprep product. DNA sequencing analysis was performed to confirm the plasmid after cloning. The 288 bp L.sericata lucimycin sequence was confirmed by DNA sequence analysis. The lucimycin gene isolated was confirmed by specific and pJET1.2 forward and reverse primers using Blastn algorithm as a result of species and/or subspecies using the Blastn algorithm and the related isolate was recorded in GenBank database with the MF964229 accessory number. The DNA sequence of the isolated sample was compared with other isolates found in GenBank by Pubmed/Blast program. KJ413251.1 was found to be 99% similar to the GenBank isolate. The 113th nucleotide was C (cytosine) in the sequence of our isolate, while the existence of G (guanine) in the sequence numbered KJ413251.1 GenBank revealed the difference between the two sequences. In this study the molecular characterization of lucimycin gene derived from L.sericata larvae were determined for the first time in Turkey, it is assumed that this molecule which has an antifungal property, can be used in the studies that will be carried out in the future, especially in microorganisms causing cutaneous infections. The study is important since the isolate is registered as a biological asset of Turkey in GenBank and also being the second study in the world.

Establishment of a Colony of Phlebotomus argentipes under Laboratory Conditions and Morphometric Variation between Wild-Caught and Laboratory-Reared Populations.

The field-based studies on sand flies are not adequate to uncover information required for the control of the leishmaniasis through reduction of vector populations. Therefore, establishment and maintenance of laboratory colonies of sand flies is an essential step in leishmaniasis research. In the current study, a colony of P. argentipes was established from wild-caught sand flies following standard procedures from the published literature. Morphological measurements of laboratory-reared and wild-caught individual sand flies were compared to assess the difference between two groups. The colony was successfully established under confined laboratory conditions. The comparison of morphometric parameters revealed that the laboratory-reared sand flies are significantly larger than those caught from wild, suggesting a possibility of increased fitness of sand flies under favorable environmental conditions which may cause higher prevalence in the disease. The current study reports the first successful attempt in colonizing sand flies under laboratory conditions. However, the colony data suggest that the conditions extracted from the published literature need to be optimized to suit local settings in order to achieve maximum population sizes within the available amount of resources.

Wolbachia Endosymbiont of the Horn Fly (Haematobia irritans irritans): a Supergroup A Strain with Multiple Horizontally Acquired Cytoplasmic Incompatibility Genes.

The horn fly, Haematobia irritansirritans, is a hematophagous parasite of livestock distributed throughout Europe, Africa, Asia, and the Americas. Welfare losses on livestock due to horn fly infestation are estimated to cost between $1 billion and $2.5 billion (U.S. dollars) annually in North America and Brazil. The endosymbiotic bacterium Wolbachia pipientis is a maternally inherited manipulator of reproductive biology in arthropods and naturally infects laboratory colonies of horn flies from Kerrville, TX, and Alberta, Canada, but it has also been identified in wild-caught samples from Canada, the United States, Mexico, and Hungary. Reassembly of PacBio long-read and Illumina genomic DNA libraries from the Kerrville H. i. irritans genome project allowed for a complete and circularized 1.3-Mb Wolbachia genome (wIrr). Annotation of wIrr yielded 1,249 coding genes, 34 tRNAs, 3 rRNAs, and 5 prophage regions. Comparative genomics and whole-genome Bayesian evolutionary analysis of wIrr compared to published Wolbachia genomes suggested that wIrr is most closely related to and diverged from Wolbachia supergroup A strains known to infect Drosophila spp. Whole-genome synteny analyses between wIrr and closely related genomes indicated that wIrr has undergone significant genome rearrangements while maintaining high nucleotide identity. Comparative analysis of the cytoplasmic incompatibility (CI) genes of wIrr suggested two phylogenetically distinct CI loci and acquisition of another cifB homolog from phylogenetically distant supergroup A Wolbachia strains, suggesting horizontal acquisition of these loci. The wIrr genome provides a resource for future examination of the impact Wolbachia may have in both biocontrol and potential insecticide resistance of horn flies.IMPORTANCE Horn flies, Haematobia irritans irritans, are obligate hematophagous parasites of cattle having significant effects on production and animal welfare. Control of horn flies mainly relies on the use of insecticides, but issues with resistance have increased interest in development of alternative means of control. Wolbachia pipientis is an endosymbiont bacterium known to have a range of effects on host reproduction, such as induction of cytoplasmic incompatibility, feminization, male killing, and also impacts vector transmission. These characteristics of Wolbachia have been exploited in biological control approaches for a range of insect pests. Here we report the assembly and annotation of the circular genome of the Wolbachia strain of the Kerrville, TX, horn fly (wIrr). Annotation of wIrr suggests its unique features, including the horizontal acquisition of additional transcriptionally active cytoplasmic incompatibility loci. This study provides the foundation for future studies of Wolbachia-induced biological effects for control of horn flies.

Sand Fly Colony Crash Tentatively Attributed to Nematode Infestation.

Maintenance of laboratory colonies of insects and other arthropod pests offers significant research advantages. The availability, age, sex, housing conditions, nutrition, and relative uniformity over time of biological material for research facilitate comparison of results between experiments that would otherwise be difficult or impossible. A laboratory research colony of Phlebotomus papatasi (Scopoli), old world sand flies, was maintained with high-colony productivity for a number of years, but within a relatively short (4-6 mo) time period, colony productivity declined from over 10,000 flies per week to less than 100 per week. Mites and nematodes were both visible in the larval medium; however, the mites had been present throughout high productivity periods; therefore, it seemed reasonable to investigate the nematodes. PCR amplification of 18S rRNA yielded a clean cDNA sequence identified by BLAST search as Procephalobus sp. 1 WB-2008 (Rhabditida: Panagrolaimidae) small subunit ribosomal RNA gene, GenBank EU543179.1, with 475/477 nucleotide identities. Nematode samples were collected and identified as Tricephalobus steineri, (Andrássy, 1952) Rühm, 1956 (Rhabditida: Panagrolaimidae) based on morphological characteristics of the esophagus and the male copulatory apparatus. Mites (Tyrophagus putrescentiae [Acariformes: Acaridae]) may have played an additional predatory role in the loss of sand fly colony productivity. We hypothesized that the origin of the nematode infestation was rabbit dung from a local rabbitry used in preparation of the larval medium. Colony productivity was fully restored within 3 mo (two sand fly generational periods) by replacement of the rabbit dung from a clean source for use to prepare sand fly larval medium.

Standardisation of quantitative resource conversion studies with black soldier fly larvae

Using larvae of the black soldier fly (Hermetia illucens; BSF) to convert low-value residual organic resources into high-value products like protein-rich animal feed ingredients and biofuel while managing organic waste has developed into a global industry. Considering the associated exponential increase in publications dealing with diet conversion efficiency by BSF larvae, it is timely to suggest procedures to arrive at an improved harmonization and reproducibility among studies. This means establishing protocols for describing the basic experiment design, fly colony origin, rearing procedures, reference and experimental feeding substrates, and sampling preparations including microbiota and chemical analyses. Such standardised protocols are instrumental to allow conversion efficiencies to be calculated. Some of these parameters are relatively easy to describe such as giving the origin and rearing conditions, while others are more challenging (e.g. description of microbe community). In this article we discuss and propose such procedures with the aim to arrive at standardisation of how future resource conversion studies with BSF larvae are conducted and how results are communicated.