Latrodectus Research Articles

Chromosome-level genome assembly of the black widow spider Latrodectus elegans illuminates composition and evolution of venom and silk proteins.

BackgroundThe black widow spider has both extraordinarily neurotoxic venom and three-dimensional cobwebs composed of diverse types of silk. However, a high-quality reference genome for the black widow spider was still unavailable, which hindered deep understanding and application of the valuable biomass.FindingsWe assembled the Latrodectus elegans genome, including a genome size of 1.57 Gb with contig N50 of 4.34 Mb and scaffold N50 of 114.31 Mb. Hi-C scaffolding assigned 98.08% of the genome to 14 pseudo-chromosomes, and with BUSCO, completeness analysis revealed that 98.4% of the core eukaryotic genes were completely present in this genome. Annotation of this genome identified that repetitive sequences account for 506.09 Mb (32.30%) and 20,167 protein-coding genes, and specifically, we identified 55 toxin genes and 26 spidroins and provide preliminary analysis of their composition and evolution.ConclusionsWe present the first chromosome-level genome assembly of a black widow spider and provide substantial toxin and spidroin gene resources. These high-qualified genomic data add valuable resources from a representative spider group and contribute to deep exploration of spider genome evolution, especially in terms of the important issues on the diversification of venom and web-weaving pattern. The sequence data are also firsthand templates for further application of the spider biomass.

Ecdysteroid responses to urban heat island conditions during development of the western black widow spider (Latrodectus hesperus).

The steroid hormone 20-hydroxyecdysone (20E) controls molting in arthropods. The timing of 20E production, and subsequent developmental transitions, is influenced by a variety of environmental factors including nutrition, photoperiod, and temperature, which is particularly relevant in the face of climate change. Environmental changes, combined with rapid urbanization, and the increasing prevalence of urban heat islands (UHI) have contributed to an overall decrease in biodiversity making it critical to understand how organisms respond to elevating global temperatures. Some arthropods, such as the Western black widow spider, Latrodectus hesperus, appear to thrive under UHI conditions, but the physiological mechanism underlying their success has not been explored. Here we examine the relationship between hemolymph 20E titers and spiderling development under non-urban desert (27°C), intermediate (30°C), and urban (33°C) temperatures. We found that a presumptive molt-inducing 20E peak observed in spiders at non-urban desert temperatures was reduced and delayed at higher temperatures. Intermolt 20E titers were also significantly altered in spiders reared under UHI temperatures. Despite the apparent success of black widows in urban environments, we noted that, coincident with the effects on 20E, there were numerous negative effects of elevated temperatures on spiderling development. The differential effects of temperature on pre-molt and intermolt 20E titers suggest distinct hormonal mechanisms underlying the physiological, developmental, and behavioral response to heat, allowing spiders to better cope with urban environments.

Risky business: males choose more receptive adults over safer subadults in a cannibalistic spider.

Understanding factors affecting male mate choice can be important for tracking the dynamics of sexual selection in nature. Male brown widow spiders (Latrodectus geometricus) mate with adult as well as immature (subadult) females. Mating with adults involves costly courtship with a repertoire of signaling behaviors, and typically ends with cannibalism (“self-sacrifice” initiated by male somersault). Mating with subadults involves brief courtship with behavioral repertoire reduced to one component (vibration) and no cannibalism. We examined male mate choice as a function of risks associated with different types of mates and the cues available to courting males. Previous studies showed male preference for adults based on air-borne pheromones, but it was unclear whether that preference is maintained after males reach female’s webs. We show that males prefer adults also based on silk-borne contact cues. To determine which types of cues trigger different courtship components, we swapped adults and subadults between webs. We showed that contact with adult females’ webs triggers two courtship behaviors from the repertoire, with adult female’s bodies triggering additional behaviors. However, vibrational signals occur regardless of the web origin or female developmental stage. We conclude that males recognize subadult females as potential mates, but are more likely to invest in costly courtship behaviors and mating attempts with adults. In our experiments, subadults were less likely to mate than adults. We conclude that mating with adults could be the preferred option for males because of the higher likelihood of copulation, even at the cost of a higher risk of cannibalism.

Differential parasitism of native and invasive widow spider egg sacs

AbstractDuring colonization, invasive species establish and spread to new locations, where they may have an advantage over native species. One such advantage may be avoidance of predators or parasites by means of better defenses or due to lower suitability as a host. We conducted field surveys and lab behavioral experiments to investigate the differential susceptibility of two widow spider species—one native to Israel, the white widow spider Latrodectus pallidus, and one invasive species, the brown widow spider, Latrodectus geometricus—to an egg sac parasitoid wasp, Philolema latrodecti. In collections of egg sacs from six paired sites of L. geometricus and L. pallidus populations in the Negev Desert, Israel, we found higher parasitism rates on the egg sacs of the native L. pallidus. In no-choice trials, we found that wasps were more likely to parasitize and oviposited longer on L. pallidus egg sacs than on L. geometricus egg sacs. In two-choice tests with spider webs and egg sacs, parasitoids made first contact with L. pallidus webs more often and faster. After developing inside of L. pallidus egg sacs, more parasitoids emerged and were larger than those emerging from L. geometricus egg sacs. Potentially better defense of the L. geometricus egg sacs as well as the parasitoid’s fitness advantages gained from parasitizing L. pallidus egg sacs may explain the higher parasitism rate in the native species. Our results suggest that the invasion and establishment success of L. geometricus is due, in part, to its ability to escape parasitism.

Dispersal and life history of brown widow spiders in dated invasive populations on two continents

Theory and empirical work suggest that behaviours such as dispersal and exploration are predictors of invasive success, and that behaviours may shift predictably after invasive populations have established and spread. However, there are limited data on temporal patterns in the distribution of behavioural traits linked to the timeline of establishment of invasive species. We examined dispersal and exploration, along with life history traits that may be linked to behaviour, across multiple invasive populations of the brown widow spider, Latrodectus geometricus. This global invader has established populations across the United States and Israel. Using this temporal and spatial variation, we tested predictions about changes in suites of traits over establishment time. We compared trait distributions of four U.S. populations of L. geometricus to patterns in four populations in Israel. We predicted that selective filters during the invasion process would result in more dispersive, more exploratory spiders that are larger and more fecund in recently established populations, but, if trade-offs occur, dispersal would be favoured at the expense of fecundity and size in recent populations. We found more frequent and faster dispersal in more recently established populations in Israel, but not the United States. Spiders in more recently established populations in Israel were larger than those in older populations, but there were no consistent patterns across U.S. populations. However, there was evidence in both the U.S. and Israel for differing trade-offs among fecundity, dispersal and size. In more recently established populations, spiders were more variable in resource allocation to eggsacs. The results suggest that in some populations, trade-offs underlying dispersal, fecundity and body size are shaped by the time interval, and thus the number of generations, since establishment, with implications for the role of evolutionary processes in invasion success.

Enhanced Black Widow Algorithm for Numerical Functions Optimization

Black widow optimization algorithm is a recently evolutionary metaheuristic that imitates the unique mating behaviour of the black widow spiders in the real life. The trend of published papers utilizing the BWO algorithm is growing rapidly due to its efficiency in solving various engineering optimization problems. However, the BWO does not always perform as well as it should, and this is due to the random initialization of the spiders also the loss of good candidate solutions during the search. To remedy these problems, we propose in this paper a modified black widow optimization algorithm (MBWO) based on three ideas. First, an efficient initialization technique is adopted, which can guarantee starting the search with finest quality spiders and plays a significant role in determining an optimal or near-optimal solution. Second, the sexual cannibalism phase is modified to avoid the loss of high-quality solutions. Finally, an adaptive adjustment of crossover and mutation probabilities is presented to achieve a compromise between the diversification and intensification. Experiments are carried out on nineteen standard benchmark functions with different dimensions. The simulation results reveal that MBWO algorithm outperforms the original one also other metaheuristic algorithms in term of solution accuracy, global optimality, and the convergence speed.

Webslinger vs. Dark Knight: First record of a false widow spider Steatoda nobilis preying on a pipistrelle bat in Britain

AbstractThe noble false widow spider Steatoda nobilis continues to expand its range globally with possible implications for public health and native species. Of the European Union list of 66 species of invasive plants and animals, none are spiders. S. nobilis is invasive around the world, but its impact on native species has not been fully assessed. Here, we report on the predation of a protected species of pipistrelle bat by S. nobilis in Britain. To our knowledge, this is the first recorded case of a Theridiidae spider preying on a bat globally, the first case of a Theridiidae spider preying on a vertebrate in Britain, and the first global case of a Steatoda preying on a mammal.

Predation strategy of the brown widow spider Latrodectus geometricus Koch, 1841 against prey with different defensive mechanisms

Predation strategy of the brown widow spider Latrodectus geometricus Koch, 1841 against prey with different defensive mechanisms

Latrotoxin-Induced Neuromuscular Junction Degeneration Reveals Urocortin 2 as a Critical Contributor to Motor Axon Terminal Regeneration.

We used α-Latrotoxin (α-LTx), the main neurotoxic component of the black widow spider venom, which causes degeneration of the neuromuscular junction (NMJ) followed by a rapid and complete regeneration, as a molecular tool to identify by RNA transcriptomics factors contributing to the structural and functional recovery of the NMJ. We found that Urocortin 2 (UCN2), a neuropeptide involved in the stress response, is rapidly expressed at the NMJ after acute damage and that inhibition of CRHR2, the specific receptor of UCN2, delays neuromuscular transmission rescue. Experiments in neuronal cultures show that CRHR2 localises at the axonal tips of growing spinal motor neurons and that its expression inversely correlates with synaptic maturation. Moreover, exogenous UCN2 enhances the growth of axonal sprouts in cultured neurons in a CRHR2-dependent manner, pointing to a role of the UCN2-CRHR2 axis in the regulation of axonal growth and synaptogenesis. Consistently, exogenous administration of UCN2 strongly accelerates the regrowth of motor axon terminals degenerated by α-LTx, thereby contributing to the functional recovery of neuromuscular transmission after damage. Taken together, our results posit a novel role for UCN2 and CRHR2 as a signalling axis involved in NMJ regeneration.

A Novel Mppt Control Strategy for Partial Shading Situations Based on the Black Widow Spider Optimization Algorithm

A Novel Mppt Control Strategy for Partial Shading Situations Based on the Black Widow Spider Optimization Algorithm

Frequency Regulation of Interconnected Power System Using Black Widow Optimization

A frequency regulation of interconnected power system comprising thermal reheat system and photovoltaic panel systems is disclosed in this research article with Proportional Integral Derivative (PID) controller optimized using Black Widow Optimization Algorithm (BWOA) based on distinct feature of cannibalism of black widow spiders. With rising demand for perfectly optimized power systems, a best optimal solution for controller gains besides merit of faster convergence and avoidance of trapping in local minimal are becoming key requirements for system engineers. In tune with these requirements, we have developed an optimized solution based on BWOA in the power system realm to minimize the fitness function which is Integral Time Absolute Error composing of frequency and tie-line power variations. Superiority of BWOA optimized controller in power system realm is established by comparing and analyzing its simulation results with some other known tuned controllers which uses different optimization algorithms namely firefly algorithm, genetic algorithm, whale optimization algorithm and modified whale optimization algorithm. Results in this paper lays out that BWOA optimized PID controller outperforms other reported controllers in terms of fitness function value, settling time, undershoot/overshoot of the incremental variations in frequency and tie-line power of the interconnected power system. Further, the potency and robustness of the proposed tuned controller are also proven by considering large variation in load demand, real-time constraints namely generation rate constraint and governor dead band in the system. Additionally, the sensitivity analysis of the optimized controlled system is performed to conclude that the proposed BWOA optimized PID controller optimized controller is insensitive to changes in system parameters and eliminates the necessity of resetting of controller parameters. The performance of the proposed control technique is also tested on two-area non-reheat thermal power system with non-linear constraints namely time delay, generation rate constraint and governor deadband.

New Record of Brown Widow Spider (Latrodectus geometricus Koch, 1841) in Ha’il Region, Saudi Arabia

Brown Widow Spider (Latrodectus geometricus) is recorded for the first time in Hail Region of Saudi Arabia .Three specimens of brown widow spider (L. geometricus ) were found and collected on October, 2021 from inside a beehive in Al Hait city (Southern of Ha’il region of Saudi Arabia). The specimens were collected and submitted to Biology department of Hail University for the identification of morphological characters and DNA barcode sequence analysis of the specimen. Phylogenetic analysis was performed to compare the DNA sequence of the newly discovered female brown widow spider with the other species of Latrodectus genus.

De Novo Transcriptome Analysis of the Venom of Latrodectus geometricus with the Discovery of an Insect-Selective Na Channel Modulator.

The brown widow spider, Latrodectus geometricus, is a predator of a variety of agricultural insects and is also hazardous for humans. Its venom is a true pharmacopeia representing neurotoxic peptides targeting the ion channels and/or receptors of both vertebrates and invertebrates. The lack of transcriptomic information, however, limits our knowledge of the diversity of components present in its venom. The purpose of this study was two-fold: (1) carry out a transcriptomic analysis of the venom, and (2) investigate the bioactivity of the venom using an electrophysiological bioassay. From 32,505 assembled transcripts, 8 toxin families were classified, and the ankyrin repeats (ANK), agatoxin, centipede toxin, ctenitoxin, lycotoxin, scorpion toxin-like, and SCP families were reported in the L. geometricus venom gland. The diversity of L. geometricus venom was also uncovered by the transcriptomics approach with the presence of defensins, chitinases, translationally controlled tumor proteins (TCTPs), leucine-rich proteins, serine proteases, and other important venom components. The venom was also chromatographically purified, and the activity contained in the fractions was investigated using an electrophysiological bioassay with the use of a voltage clamp on ion channels in order to find if the neurotoxic effects of the spider venom could be linked to a particular molecular target. The findings show that U24-ctenitoxin-Pn1a involves the inhibition of the insect sodium (Nav) channels, BgNav and DmNav. This study provides an overview of the molecular diversity of L. geometricus venom, which can be used as a reference for the venom of other spider species. The venom composition profile also increases our knowledge for the development of novel insecticides targeting voltage-gated sodium channels.

Urban Environments Aid Invasion of Brown Widows (Theridiidae: Latrodectus geometricus) in North America, Constraining Regions of Overlap and Mitigating Potential Impact on Native Widows

Urbanization is a major cause of biotic homogenization and habitat fragmentation for native communities. However, the role of urbanization on the success of biological invasions on a continental scale has yet to be explored. Urbanization may facilitate the establishment success of invasive species by minimizing niche differentiation between native and invaded ranges. In such cases, we might expect anthropogenic variables to have stronger influence on the geographic distribution of invasive compared to native populations. In this study, we use ecological niche modeling to define the distribution of non-native brown widow spider (Latrodectus geometricus) and three native black widows (L. hespersus, L. mactans, L. variolus) in North America and gauge the importance of urbanization on the geographic ranges of widows at a continental scale. We also quantify the geographic overlap of L. geometricus with each native widow to assess potential species and regions at risk of ecological impact. Consistent with our hypothesis, we find that the distribution of L. geometricus is strongly constrained to urban environments, while native widow distributions are more strongly driven by climatic factors. These results show that urbanization plays a significant role in facilitating the success of invasion, weakening the significance of climate on the realized niche in its invaded range.

Molecular architecture of black widow spider neurotoxins

Latrotoxins (LaTXs) are presynaptic pore-forming neurotoxins found in the venom of Latrodectus spiders. The venom contains a toxic cocktail of seven LaTXs, with one of them targeting vertebrates (α-latrotoxin (α-LTX)), five specialized on insects (α, β, γ, δ, ε- latroinsectotoxins (LITs), and one on crustaceans (α-latrocrustatoxin (α-LCT)). LaTXs bind to specific receptors on the surface of neuronal cells, inducing the release of neurotransmitters either by directly stimulating exocytosis or by forming Ca2+-conductive tetrameric pores in the membrane. Despite extensive studies in the past decades, a high-resolution structure of a LaTX is not yet available and the precise mechanism of LaTX action remains unclear. Here, we report cryoEM structures of the α-LCT monomer and the δ-LIT dimer. The structures reveal that LaTXs are organized in four domains. A C-terminal domain of ankyrin-like repeats shields a central membrane insertion domain of six parallel α-helices. Both domains are flexibly linked via an N-terminal α-helical domain and a small β-sheet domain. A comparison between the structures suggests that oligomerization involves major conformational changes in LaTXs with longer C-terminal domains. Based on our data we propose a cyclic mechanism of oligomerization, taking place prior membrane insertion. Both recombinant α-LCT and δ-LIT form channels in artificial membrane bilayers, that are stabilized by Ca2+ ions and allow calcium flux at negative membrane potentials. Our comparative analysis between α-LCT and δ-LIT provides first crucial insights towards understanding the molecular mechanism of the LaTX family.

Don't put all your eggs in small baskets: Ineffective guardians, incidence of parasitoids and clutch size of Latrodectus geometricus (Araneae, Theridiidae) along an urban gradient

Maternal care is common in spiders, but the effects of egg protection by mothers on offspring survival remains understudied for most species, especially against parasitoids. Parasitoid incidence can also be modulated by host clutch size (since larger egg masses provide the opportunity of producing larger clutches for the parasitoids) and urbanization (by creating conditions that may restrict the occurrence of less tolerant species). In this study, we conducted field experiments with the brown widow spider Latrodectus geometricus to investigate the following hypothesis: (1) females are effective in the defense of eggs against parasitoids; (2) large clutch sizes are attacked more often by parasitoids than the smaller ones; and (3) the incidence of parasitoids in egg sacs decreases in a gradient from a preserved area to an urbanized one. The presence of caring females reduced by three times the incidence of two parasitoids, Baeus latrodecti (Hymenoptera, Scelionidae) and an unidentified Eulophidae wasp, compared to unattended egg sacs, but this distinction did not differ from a random distribution. Parasitoids were most often found in egg sacs containing few eggs. The incidence of B. latrodecti in egg sacs was highest in the area with an intermediate degree of urbanization, but Eulophidae wasps were unaffected by the urbanization gradient. We concluded that (1) females are generally ineffective at protecting their egg sacs against parasitoids, that (2) egg sacs containing small clutch sizes are usually more vunerable to parasitoid attacks, and that (3) urbanization does not reduce the incidence of the egg parasitoids identified here. Our study provides an important step towards our understanding of the interactions between spiders with parental care and egg parasitoids, suggesting new aspects to be investigated in future studies.

Expression of Brown and Southern Black Widow Spider (Araneae: Theridiidae) Latrotoxins Is Tissue- and Life Stage-Specific for α-Latroinsectotoxins and δ-Latroinsectotoxins and Is Ubiquitous for α-Latrotoxins.

Widow spiders are widely known for their potent venom toxins that make them among the few spiders of medical concern. The latrotoxins are the most well-studied widow toxins and include both the vertebrate-specific latrotoxins and the insect-specific latroinsectotoxins (LITs). Previous studies have shown that toxins are not limited to expression in the venom glands of adult spiders; however, gaps exist in latrotoxin screening across all life stages for brown widows, Latrodectus geometricus and southern black widows, Latrodectus mactans. In this study, we screened male and female venom gland, cephalothorax, and abdomen tissues, spiderling cephalothorax and abdomen tissues, and eggs of both L. geometricus and L. mactans, for the presence of three latrotoxins: α-latrotoxin (α-LTX), and α- and δ-latroinsectotoxins (α/δ-LITs). Widows were locally collected. Extracted RNA was used to prepare cDNA that was analyzed by PCR for the presence or absence of latrotoxin expression. Results show that expression profiles between the two species are very similar but not identical. Expression of α-LTX was found in all life stages in all tissues examined for both species. For both species, no LIT expression was detected in eggs and variable patterns of α-LIT expression were detected in spiderlings and adults. Notably, δ-LIT could only be detected in females for both species. Our results show that latrotoxin expression profiles differ within and between widow species. Data on their expression distribution provide further insight into the specific latrotoxins that contribute to toxicity profiles for each life stage in each species and their specific role in widow biology.

Dry-Heat Tolerance of Egg Sacs of Invasive Latrodectus Spiders (Araneae: Theridiidae) in Japan: Implications for Efficient Control/Extermination.

Strategic responses to invasive Latrodectus widow spiders are a global challenge due to the risks they pose to health and ecosystems. Chemical strategies involving the use of pyrethroids are effective against adult spiders, but as their populations rebound, chemical control becomes costly and unsustainable for eradication. A major obstacle is the inefficacy of insecticides against eggs, which are covered by a protective silk egg sac. Eradication of invasive spiders must focus on destroying progeny. Here, the responses of eggs in egg sacs of two invasive Latrodectus spiders in Japan (Latrodectus hasseltii (Thorell) and Latrodectus geometricus (C.L. Koch)) to short-term dry-heat exposure were examined. To test whether the dry-heat tolerance of the egg sacs of both spider species differed, lethal temperature (LT) was determined based on the hatching rate of eggs from egg sacs subjected to a range of temperatures. Hatching in both species failed completely when the egg sacs were exposed to temperatures of 55°C and above for 10 min, but the LT to reduce hatching by 90% (LT90) differed significantly between L. hasseltii (50. 9°C) and L. geometricus (52. 8°C). Our study highlights the efficacy of dry heat in suppressing hatching and thus shows the possibility for effective extermination of these noxious invasive pests. Further exploration and investigation of the effects of humidity and heat exposure time on egg sacs under field conditions are needed to guide Latrodectus spider control strategies.

Two-Screen Virtual Board Game Didactic for Teaching Wilderness and Environmental Medicine Topics to Emergency Medicine Residents.

This game is appropriate for medical students, interns, junior and senior residents. The COVID-19 pandemic has forced a transition from in-person to virtual learning, and educators must innovate and adapt to keep learners engaged. One way to achieve this is through gamification.1 The authors employed a novel approach to gamification of virtual learning which engaged not only learners' computers but also their mobile phones. Learners worked in teams communicating by text message to answer ABEM board-style questions and occupy sites on the virtual board. By the end of this didactic, the learner will:1. Describe the basics of the presentation of each topic listed above2. Recall the basics of management of each topic listed above3. Improve learners' preparedness for the Emergency Medicine Inservice Exam and Written Board Examination. Wilderness and environmental medicine topics were selected from the list of topics covered on the ABEM boards. Questions were then written by the authors teaching these concepts. Learners were surveyed immediately following the session using an evaluation tool containing both Likert-scale questions on quality and applicability as well as open-ended questions on strengths and areas for improvement. The response rate to this survey was 100%. Feedback was overwhelmingly positive, with 19/20 respondents rating the sessions 5/5 for effectiveness and value of teaching compared with other sessions, and 1/20 rating the session 4/5. Nineteen out of twenty respondents rated the content as "highly relevant"; 1/20 rated it as "mostly relevant." Nineteen out of twenty respondents rated the session 5/5 for being engaging and holding their attention; 1/20 rated it as somewhat engaging. Learners rated the session as highly relevant and engaging. We hypothesize that by engaging two screens and forcing learners to work together by text message, we were able to turn what would normally be a distraction (texting co-residents during resident conference) into a tool for learning. Electrical injury, lightning strike, thermal burns, inhalational injury, chemical burns, acute radiation syndrome, snake bites, scorpion envenomation, stingray envenomation, jellyfish stings, black widow spider bites, mammalian bites, rabies, murine typhus, bear encounters, decompression sickness, arterial gas embolism, drowning, dehydration, heat stroke, exercise-associated hyponatremia, frostbite, hypothermia, CO poisoning, hydrogen sulfide poisoning, giardia.

Real time flood disaster monitoring based on energy efficient ensemble clustering mechanism in wireless sensor network

AbstractIn this article, energy efficient ensemble clustering method (EECM) with black widow optimization (EECM‐BWO) algorithm is proposed for effective data transmission with the help of real time flood disaster monitoring wireless sensor network (WSN). Initially, unified scalable ensemble clustering algorithm based on ensemble generation and consensus function is proposed for selecting the optimal routing path among the node using BWO algorithm. Then, biologically inspired routing black widow spiders optimization algorithm is proposed to trade off the nodes energy level, self‐organization, and self‐configuration in the WSN. The simulation is performed using NS2 simulator for validating the performance of the proposed EECM‐BWO method. Here, in node, low delay achieves 24.07%, 72.58%, 51.36%, 81.75%, 77.74%, high packet delivery ratio achieves 70.83%, 53.93%, 90.23%, 43.58%, 24.58%, low packet drop attains 77.93%, 72.76%, 61.56%, 51.87%, 34.35%, low energy consumption attains 75.9%, 52.94%, 65.81%, 58%, 41.2% compared with existing energy‐efficient clustering approach consolidated game theory as well as dual‐cluster‐head mode for WSNs energy‐aware clustering by cuckoo optimization approach (EECM‐COA), energy‐aware clustering‐based routing using multi‐path reliable transmission with routing and control board (EECM‐RCB‐MRT), adaptive repair algorithm with temporally ordered routing algorithms for flood control strategy (EECM‐AR‐TORA‐FCS), passive multi‐hop clustering algorithm (EECM‐PMC), dynamic source routing protocol based on genetic algorithm‐bacterial foraging optimization (DSR‐GA‐BFO).