Hygienic behavior in Melipona quadrifasciata anthidioides (Apidae, Meliponini) Jossimara Neiva de Jesus, Emerson Dechechi Chambó, Geni da Silva Sodré, Newton Tavares Escocard de Oliveira, Carlos Alfredo Lopes de Carvalho To cite this version: Jossimara Neiva de Jesus, Emerson Dechechi Chambó, Geni da Silva Sodré, Newton Tavares Escocard de Oliveira, Carlos Alfredo Lopes de Carvalho. Hygienic behavior in Melipona quadrifasciata anthid- ioides (Apidae, Meliponini). Apidologie, Springer Verlag, 2017, 48 (4), pp.504-512. 10.1007/s13592- 017-0495-5. hal-01710374 HAL Id: hal-01710374 https://hal.archives-ouvertes.fr/hal-01710374 Submitted on 15 Feb 2018 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. 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Apidologie (2017) 48:504–512 Original article * INRA, DIB and Springer-Verlag France, 2017 DOI: 10.1007/s13592-017-0495-5 Hygienic behavior in Melipona quadrifasciata anthidioides (Apidae, Meliponini) 1 1 1 Jossimara Neiva de JESUS , Emerson Dechechi CHAMBÓ , Geni da SILVA SODRÉ , 2 1 Newton Tavares Escocard de OLIVEIRA , Carlos Alfredo Lopes de CARVALHO 1Universidade Federal do Recôncavo da Bahia, Rua Rui Barbosa 710, CEP 44380-000, Cruz das Almas, Bahia, Brazil 2Universidade Estadual do Oeste do Paraná, Rua Pernambuco 1777, CEP 45980-000, Marechal Cândido Rondon, Paraná, Brazil Received 3 October 2016 – Revised 30 December 2016 – Accepted 16 January 2017 Abstract – Hygienic behavior in stingless bees is a trait of workers that confers colony-level resistance against some brood diseases. Workers of hygienic colonies detect, uncap and remove dead or diseased brood from the nest cells. We examined the hygienic behavior in stingless bees (Melipona quadrifasciata anthidioides ) from freeze- killed brood assay using liquid nitrogen. Responses were measured at 14 times (3, 6, 12, 24, 48, 72, 96, 120, 144, 168, 192, 216, 240 and 264 h after freeze-killing of the brood). Workers were estimated to remove on average 65% of larvae and 34% of dead pupae within 48 h of freezing. Workers removed dead brood rapidly after uncapping the cells. Strong colonies showed a greater removal of dead pupae, while the size of the population did not influence the removal of dead larvae. These findings report for the first time the hygienic behavior in M. q. anthidioides and confirm that workers have more difficulty removing pupae compared with larvae from the combs. generalized linear models / stingless bees / hygienic colonies / dead brood removal / freeze-killed brood 1. INTRODUCTION 2010) and populations of stingless bees have de- creased worryingly (Steffan-Dewenter et al. 2005; Melipona quadrifasciata Lepeletier Slaa et al. 2006; Freitas et al. 2009). There are (Bmandaçaia^) is a stingless bee species that oc- numerous reasons for these reductions, such as the curs in most of Brazil, from the states of Paraíba to destruction or changes of habitats, pesticide overuse Rio Grande do Sul (Moure and Kerr 1950). The in agricultural crops, and diseases that affect bees higher prevalence of subspecies (vanEngelsdorp and Meixner 2010;Reyes- M. quadrifasciata anthidioides is in hot and dry González et al. 2016; Sanchez-Bayo and Goka regions, especially in the semi-arid region of the 2016 and references therein). In the case of diseases, state of Bahia (Nunes et al. 2008). Honey pro- a practical and desirable strategy is that the bee duced by M. q. anthidioides is highly appreciated species express high levels of natural resistance, and its colonies present good production when such as the hygienic behavior (HB) (Evans and managed rationally, being of great socioeconomic Spivak 2010; Bigio et al. 2013). and environmental importance. In A. mellifera , HB has been described as a In recent decades, bee colonies of Apis mellifera process in which bees detect and uncap dead, (vanEngelsdorp et al. 2007; Neumann and Carreck parasitized or infected brood cells (5th instar, lar- vae and pupae) and remove them from the nest (Rothenbuhler 1964; Rosenkranz et al. 2010). Corresponding author: E. Chambó, When quickly performed, this process may pre- [email protected] vent the spread of the disease causal agents to Manuscript editor: James Nieh healthy brood (Rothenbuhler 1964). HB is a Hygienic behavior in bees 505 significant general resistance mechanism to a boxes, INPA model, consisting of first deep and second number of pathogens, including Ascosphaera deep (13 × 13 × 6 cm), honey super (13 × 13 × 3.5 cm), apis (causal agent of chalk brood) (Gilliam et al. outer cover and bottom board (18 × 18 × 2 cm) and 1988), Paenibacillus larvae (causal agent of internal space to access the compartments (6 × 6 cm). American foulbrood) (Spivak and Reuter 2001) HB was quantified at the level of the colony using and the parasitic mite Varroa destructor (Rinderer the freezing method with liquid nitrogen (N2) et al. 2010;Rasolofoarivaoetal.2015). (described in Spivak and Reuter 1998), adapted to the Hygienic colonies have large economic inter- nest dimensions of the species under study. For that ests in apiculture, because they have been reported purpose, an area with seven cells containing black- to produce more honey and pollen than non- eyed pupae and seven larvae in the last instar (both hygienic colonies (Nicodemo et al. 2013). Al- capped cells) was delimited and marked. A control area though HB is a very important feature, it rarely was also delimited and marked to discount the natural manifests with intensity in populations (Pérez- removal rate of diseased, dead or parasitized pupae and Sato et al. 2009). Thus, the selection of queen larvae. A PVC cylinder (2 cm diameter × 3 cm high) bees related to this characteristic has been intensi- was positioned over the areas to be tested and 12 mL of fied in breeding programs (Espinosa-Montaño liquid nitrogen were poured to kill the brood. Each et al. 2008; Costa-Maia et al. 2011). colony was monitored until the worker bees performed Most studies on HB are carried out with the uncapping and totally removed the brood. A. mellifera (Gramacho and Gonçalves 2009; The experiment was installed in a complete random- Pérez-Sato et al. 2009; Wilson-Rich et al. 2009; ized block design with 14 treatments repeated 40 times Morais et al. 2010; Stevanovic et al. 2011; Pinto (colonies) in each block, totaling 1680 records. The et al. 2012;Bigioetal.2013; Nicodemo et al. 2013; blocks were made of three different observation periods Pereira et al. 2013; Rasolofoarivao et al. 2015). Few for each colony, with intervals of 30 days between each species of stingless bees have been investigated in observation. The treatments or measurement times oc- terms of HB, namely Melipona beecheii , curred up to 264 h, as follows, 3, 6, 12, 24, 48, 72, 96, Scaptotrigona pectoralis (Medina et al. 2009)and 120, 144, 168, 192, 216, 240 and 264 h after liquid Plebeia remota (Nunes-Silva et al. 2009). nitrogen application. The variables analyzed were: ac- Therefore, HB investigations in other species cumulated average proportion of dead larvae removal of stingless bees is necessary, mainly because of (LR) and dead pupae removal (PR), and accumulated the diversity and specific characteristics of each average proportion of uncapped cells with not yet re- species, extinction risks for the Meliponini and the moved dead larvae (UL) and pupae (UP). All the colo- growth of meliponiculture which can increase the nies assessed for HB had the population size estimated risks of pathogen transmission (Venturieri et al. before each repetition according to Ihering (1932)and 2012), especially to species of greater economic after measuring the number of brood cells (Aidar 2010). interest. In addition, no study on HB in M. q. Generalized linear models (GLM) with binomial anthidioides has been conducted. error structure and logit link function expressed by g Our hypothesis is that HB in M. q. anthidioides (μ) = ln (μ/1-μ) were used to test the effect of time and occurs in a similar manner to that of bees of the population of the colonies on HB. The model parame- genus Apis and that there is difference in the ters were estimated using the method of maximum removal time of dead larvae and pupae by worker likelihood by maximizing the log-likelihood function bees. The objective was to use M. q. anthidioides through generalized estimating equations (GEE). The as a model to investigate HB in short time intervals GEE considered the dependence between the observa- until the complete removal of all the brood (larvae tions measured within the times (hours), which charac- and pupae) killed by freezing in the colonies. terized repeated measures in the experimental units (colonies). The responses obtained between experimen- 2. MATERIAL AND METHODS tal units from different periods (blocks) were assumed to be statistically independent. The deviance analysis The study was conducted from May to August, 2015 (ANODE) was used for the GLM adjustment, from a on 40 colonies of M. q. anthidioides housed in standard maximal model, represented by systematic portion 506 J.N. de Jesus et al. η =g(μ)=μ +Ti +Bj +P+εij, where μ is the general 3. RESULTS average effect, Ti is the time levels effect in hours (i = 1, 2, …,14),Bj is the blocks effect (j = 1, 2 and 3), P is the We used recommendations of Spivak and effect of the ‘colonies population’ covariate ,and εij is Downey (1998)inA. mellifera for the parameters the random error. to compare HB of the colonies of M.