Genetic markers associated with insecticide resistance and resting behaviour in Anopheles gambiae mosquitoes in selected sites in Kenya

Background Molecular diagnostic tools have been incorporated in insecticide resistance monitoring programmes to identify underlying genetic basis of resistance and develop early warning systems of vector control failure. Identifying genetic markers of insecticide resistance is crucial in enhancing the ability to mitigate potential effects of resistance. The knockdown resistance (kdr) mutation associated with resistance to DDT and pyrethroids, the acetylcholinesterase-1 (ace-1R) mutation associated with resistance to organophosphates and carbamates and 2La chromosomal inversion associated with indoor resting behaviour, were investigated in the present study. Methods Anopheles mosquitoes sampled from different sites in Kenya and collected within the context of malaria vector surveillance were analysed. Mosquitoes were collected indoors using light traps, pyrethrum spray and hand catches between August 2016 and November 2017. Mosquitoes were identified using morphological keys and Anopheles gambiae sensu lato (s.l.) mosquitoes further identified into sibling species by the polymerase chain reaction method following DNA extraction by alcohol precipitation. Anopheles gambiae and Anopheles arabiensis were analysed for the presence of the kdr and ace-1R mutations, while 2La inversion was only screened for in An. gambiae where it is polymorphic. Chi-square statistics were used to determine correlation between the 2La inversion karyotype and kdr-east mutation. Results The kdr-east mutation occurred at frequencies ranging from 0.5 to 65.6% between sites. The kdr-west mutation was only found in Migori at a total frequency of 5.3% (n = 124). No kdr mutants were detected in Tana River. The ace-1R mutation was absent in all populations. The 2La chromosomal inversion screened in An. gambiae occurred at frequencies of 87% (n = 30), 80% (n = 10) and 52% (n = 50) in Baringo, Tana River and Migori, respectively. A significant association between the 2La chromosomal inversion and the kdr-east mutation was found. Conclusion The significant association between the 2La inversion karyotype and kdr-east mutation suggests that pyrethroid resistant An. gambiae continue to rest indoors regardless of the presence of treated bed nets and residual sprays, a persistence further substantiated by studies documenting continued mosquito abundance indoors. Behavioural resistance by which Anopheles vectors prefer not to rest indoors may, therefore, not be a factor of concern in this study’s malaria vector populations.

Use of space, active and resting behaviour in fattening rabbits (Oryctolagus cuniculus) housed in a combi park system: A case study

In this case study, a housing system for rabbits (Oryctolagus cuniculus) was developed, complying with the new German Welfare Regulation, and evaluated on-farm with regard to the rabbits' behaviour during four fattening periods. The housing system was characterised by the large group size of fatteners (maximum 65 animals per group, 12 animals per m²) due to the merging of six former single units for does and their litters, post-weaning. A large elevated platform, a box with a roof (small elevated platform), a tube and gnawing materials were made available per unit. The aim being to assess the suitability of the housing system for rabbits with regard to animal welfare, based on behavioural analyses. Therefore, the use of space by the rabbits (n = 247) was investigated by video analysis (instantaneous scan sampling) during daytime. Additionally, the individual behaviour of 20 focal rabbits in different locations was assessed by continuous sampling. Results showed that rabbits preferred to huddle together in the outer units in the first weeks post-weaning. The highest animal densities were found under and in front of the large and on the small platforms. The large platforms were visited increasingly from the 10th fattening day onwards. Elevated platforms supported resting and comfort behaviours. Nonelevated open-top areas enabled upright positions and locomotor behaviours. Aggression and stereotypic behaviours rarely occurred. We conclude that the housing system supported species-specific behaviour and seemed to cater for the needs of rabbits in terms of welfare. Nevertheless, further investigation is needed to ascertain the effects on animal health and performance.

Australian mosquito assemblages vary between ground and sub-canopy habitats

Background The surveillance and control of mosquito-borne diseases is dependent upon understanding the bionomics and distribution of the vectors. Most studies of mosquito assemblages describe species abundance, richness and composition close to the ground defined often by only one sampling method. In this study, we assessed Australian mosquito species near the ground and in the sub-canopy using two traps baited with a variety of lures. Methods Mosquitoes were sampled using a 4 × 4 Latin square design at the Cattana Wetlands, Australia from February to April 2020, using passive box traps with octenol and carbon dioxide and three variations of a sticky net trap (unbaited, and baited with octenol or octenol and carbon dioxide). The traps were deployed at two different heights: ground level (≤ 1 m above the ground) and sub-canopy level (6 m above the ground). Results In total, 27 mosquito species were identified across the ground and sub-canopy levels from the different traps. The abundance of mosquitoes at the ground level was twofold greater than at the sub-canopy level. While the species richness at ground and sub-canopy levels was not significantly different, species abundance varied by the collection height. Conclusions The composition of mosquito population assemblages was correlated with the trap types and heights at which they were deployed. Coquillettidia species, which prefer feeding on birds, were mainly found in the sub-canopy whereas Anopheles farauti, Aedes vigilax and Mansonia uniformis, which have a preference for feeding on large mammals, were predominantly found near the ground. In addition to trap height, environmental factors and mosquito bionomic characteristics (e.g. larval habitat, resting behaviour and host blood preferences) may explain the vertical distribution of mosquitoes. This information is useful to better understand how vectors may acquire and transmit pathogens to hosts living at different heights. Graphical abstract

Is outdoor-resting behaviour in malaria vectors consistent? Short report from northern Ghana.

Background Recent reports of a change in the resting behaviour of malaria vectors, from predominantly indoor resting to outdoor resting following blood feeding, have been attributed to selection pressure from use of vector control tools such as indoor residual spraying and long-lasting insecticide-treated nets. Recent studies have observed vectors resting predominantly outdoors in settings where anti-vector tools are extensively deployed. This present study examined if the outdoor resting behaviour in the vector population, is random or indicative of a consistent preference of one resting site over the other. Methods Mark-release-recapture (MRR) experiments were conducted with outdoor-resting Anopheles gambiae and An. funestus mosquitoes collected from pit shelters, animal houses and granaries in two villages in Northern Ghana during rainy and dry seasons. Mosquitoes were marked with fluorescent dyes and released indoors. The experiments were controlled with indoor-resting mosquitoes, which were marked and released outdoors. Species of all recaptured mosquitoes were identified and assessed for consistency in their resting behaviour. Results A total of 4,460 outdoor-resting mosquitoes comprising An. gambiae sensu lato (s.l.) (2,630, 59%) and An. funestus complex (1,830, 41%) were marked and released. Overall, 31 (0.7%) mosquitoes were recaptured mostly from outdoor location comprising 25 (81%) An. gambiae s.l. and 6 (19%) An. funestus complex. Only 3 (10%) of the recaptured mosquitoes were found resting indoors where they were released. The majority of the outdoor-recaptured mosquitoes were An. arabiensis (11, 39%), followed by An. coluzzii (7, 25%); whereas all indoor-recaptured mosquitoes were An. coluzzii. For the control experiment, 324 indoor-resting mosquitoes constituting 313 (97%) An. gambiae s.l. and 11 (3%) An. funestus complex were marked and released. However, none of these was recaptured neither indoors nor outdoors. More mosquitoes were significantly captured and recaptured during rainy season ( Z = 6.579, P < 0.0001). Conclusions The results obtained suggested the tendency for the mosquitoes to retain their outdoor-resting behaviour. Further investigations are required to ascertain if emerging preference for outdoor resting behaviour in malaria vector populations is consistent or a random occurrence.

The “right” side of sleeping: laterality in resting behaviour of Aldabra giant tortoises (Aldabrachelys gigantea)

Although some studies investigated lateralization in reptiles, little research has been done on chelonians, focusing only on few behaviours such as righting response and escape preference. The aim of this study was to investigate lateralization in Aldabra giant tortoises (Aldabrachelys gigantea), focusing on asymmetrical positioning of the limbs and the head during resting behaviour, called sleep-like behaviour, involving both wild tortoises and individuals under human care. Subjects of the study were 67 adult Aldabra tortoises (54 free ranging on Curieuse, 13 under human care in Mahè Botanical Garden). For each tortoise observed during sleep-like behaviour, we recorded the position of the head (on the left, on the right or in line with the body midline) and we collected which forelimb and hindlimb were kept forward. Moreover, the number of subjects in which limbs were in a symmetrical position during the sleep-like behaviour was recorded. Based on our results, the number of tortoises with asymmetrical position of head and limb was higher (head: 63%; forelimbs: 88%; hindlimbs: 70%) than the number of tortoises with symmetrical position of the head and the limb. Regarding the head, throughout the subjects found with the asymmetrical position of the head during sleep-like behaviour, tortoises positioning the head on the right (42%) were more than those sleeping with the head on the left (21%). We found a relationship between the position of the forelimbs and hindlimbs during sleep-like behaviour. We reported no differences between Mahè (under human care) and Curieuse (wild) tortoises. Findings of this preliminary study underlined traces of group-level lateralization in head positioning during the sleep-like behaviour, possibly due to a left-eye/right-hemisphere involvement in anti-predatory responses and threatening stimuli as reported in reptiles and other vertebrates. This study aims at adding data on brain lateralization, often linked to lateralized behaviours, in reptiles, especially in chelonians.

Is Outdoor-Resting Behaviour in Malaria Vectors Consistent? Short Report From Northern Ghana.

BackgroundRecent reports of a change in the resting behaviour of malaria vectors, from predominantly indoor resting to outdoor resting following blood feeding, have been attributed to selection pressure from use of vector control tools such as indoor residual spraying and long-lasting insecticide-treated nets. Recent studies have observed vectors resting predominantly outdoors in settings where anti-vector tools are extensively deployed. This present study examined if the outdoor resting behaviour in the vector population, is random or indicative of a consistent preference of one resting site over the other. MethodsMark-release-recapture (MRR) experiments were conducted with outdoor-resting Anopheles gambiae and An. funestus mosquitoes collected from pit shelters, animal houses and granaries in two villages in Northern Ghana. Mosquitoes were marked with fluorescent dyes and released indoors. The experiments were controlled with indoor-resting mosquitoes, which were marked and released outdoors. Species of all recaptured mosquitoes were identified and assessed for consistency in their resting behaviour. ResultsA total of 4,460 outdoor-resting mosquitoes comprising An. gambiae sensu lato (s.l.) (59%) and An. funestus complex (41%) were marked and released. Overall, 31 (0.7%) mosquitoes (25 An. gambiae s.l. and 6 An. funestus complex) were recaptured mostly from outdoor location. Only 3 of the recaptured mosquitoes were found resting indoors where they were released. The majority of the outdoor-recaptured mosquitoes were An. arabiensis (11, 39%), followed by An. coluzzii (7, 25%); whereas the indoor-recaptured mosquitoes were mainly (3) An. coluzzii. For the control experiment, 324 indoor-resting mosquitoes constituting 313 An. gambiae s.l. and 11 An. funestus complex were marked and released. However, none of these was recaptured neither indoors nor outdoors.ConclusionsThe mosquitoes demonstrated the tendency to retain their outdoor-resting behaviour. Further investigations are required to ascertain if emerging preference for outdoor resting behaviour in malaria vector populations is consistent or a random occurrence.

Hanging by a thread: unusual nocturnal resting behaviour in a jumping spider

Background For diurnal animals that heavily rely on vision, a nocturnal resting strategy that offers protection when vision is compromised, is crucial. We found a population of a common European jumping spider (Evarcha arcuata) that rests at night by suspending themselves from a single silk thread attached overhead to the vegetation, a strategy categorically unlike typical retreat-based resting in this group. Results In a comprehensive study, we collected the first quantitative field and qualitative observation data of this surprising behaviour and provide a detailed description. We tested aspects of site fidelity and disturbance response in the field to assess potential functions of suspended resting. Spiders of both sexes and all developmental stages engage in this nocturnal resting strategy. Interestingly, individual spiders are equally able to build typical silk retreats and thus actively choose between different strategies inviting questions about what factors underlie this behavioural choice. Conclusions Our preliminary data hint at a potential sensory switch from visual sensing during the day to silk-borne vibration sensing at night when vision is compromised. The described behaviour potentially is an effective anti-predator strategy either by acting as an early alarm system via vibration sensing or by bringing the animal out of reach for nocturnal predators. We propose tractable hypotheses to test an adaptive function of suspended resting. Further studies will shed light on the sensory challenges that animals face during resting phases and should target the mechanisms and strategies by which such challenges are overcome.