AIMS Microbiology, 4(2): 319–333. DOI: 10.3934/microbiol.2018.2.319 Received: 24 January 2018 Accepted: 02 April 2018 Published: 20 April 2018 http://www.aimspress.com/journal/microbiology Research article A study on the potential of ants to act as vectors of foodborne pathogens Leckranee Simothy, Fawzi Mahomoodally and Hudaa Neetoo* Department of Agricultural and Food Sciences, Faculty of Agriculture, University of Mauritius, Réduit, Moka, 80837, Mauritius * Correspondence: Email: [email protected]; Tel: +2304037885; Fax: +2304655743. Abstract: Ants (Technomyrmex difficilis and Solenopsis geminata) are insects often found in domestic kitchens of Mauritius. Unfortunately, they harbour disease-causing organisms and can potentially transfer these pathogens to food. This study was carried out to (i) investigate the knowledge, perception and behaviors of consumers in relation to the problem of ant infestation of domestic kitchens; (ii) identify the pathogenic microorganisms carried by ants; and (iii) determine the potential for ants to transfer these pathogenic microorganisms to food. A survey based on a stratified sampling design was carried out with 100 consumers using a questionnaire. To identify the pathogenic microorganism(s) harbored by ants, bait traps were set up using sterile sugar as a non-toxic attractant. Captured ants were then subjected to microbiological analyses. Most respondents (72%) agreed that ants constitute a hygienic issue but they did not perceive ants as a serious threat to human health. However microbiological analyses of ants (n = 50) confirmed the presence of various pathogenic microorganisms as well as fecal contaminants. Ants were found to harbor yeasts and molds systematically (100%), coliforms frequently (52%), Bacillus spp. and Escherichia coli occasionally (26% and 18% respectively) and Salmonella and Listeria monocytogenes sporadically (8 and 6 % respectively). Ants were also found to transfer E. coli to food surfaces at a moderately high frequency of 70%. This study demonstrated that the majority of consumers acknowledged the problem of ant infestation as a sanitation-related problem rather than a food safety issue. Since ants have the ability to harbor and subsequently transfer pathogenic or toxigenic microorganisms, ants may act as disease vectors and contaminate food, water and food- contact surfaces of kitchens resulting in foodborne illnesses. 320 Keywords: ants; food; pathogens; consumers; vectors 1. Introduction Ants (Formicidae, Hymenoptera) are ubiquitous social insects that live in colonies. Female worker ants typically leave the nest and venture out in search for food [1]. Ants are generally omnivorous and the animal proteins and fats in their diet are derived mostly from insects and other arthropods that fall prey to the foraging worker ants [1]. Ants also feed on sugars, starches or foods containing those carbohydrates. As a result, kitchens, bakeries, restaurants, and food factories are typical sites for foraging ants [2]. Although ants are significant vectors of infectious diseases, there is considerably less information on ants than on other insect pests such as flies and cockroaches. Unfortunately, ants harbor various species of internal [3,4] as well as external [1,5,6] bacteria, which adhere to the external surfaces, mainly the legs and mandibles [1,5,6]. These appendages come into contact with substrates, such as soil and pit latrines outdoors and, most commonly, floors indoors, from which the ants may pick up pathogens [1]. As ants forage over clean food-contact surfaces, such as dishes and cutting boards, pathogens may be deposited and eventually become mixed in with a ready-to-eat food intended for human or animal consumption. Literature has indicated that multiple pathogens have been isolated from pest ants including Bacillus cereus [7,8], Clostridium perfringens [7], Escherichia coli [7], filamentous fungi [8], Klebsiella pneumoniae [9], Micrococcus sp. [8], Proteus mirabilis [9], Pseudomonas spp. [9], Salmonella sp. [7], Staphylococcus aureus [7], other Staphylococcus spp. [8] and Streptococcus pyogenes [7]. Unfortunately, ants have the potential to carry certain microorganisms to food establishments and transfer pathogenic microorganisms to food [6]. According to WHO [10], food safety is an essential aspect in public health and foodborne disease outbreaks and epidemics have been classified as a major global public health issue in the 21st century. As ants are major mechanical vectors of pathogens [11], including foodborne pathogens [8], it is important to exterminate them in kitchens and other food preparation facilities so as to prevent contamination of food. The problem of ant infestation can be addressed by preventing their ingress into homes [12]. This can be achieved by filling or sealing crevices and cracks, cleaning around common entry points with a detergent to remove the chemical trail of pheromone along their routes to and from a food source, or using a non-repellent residual insecticide [13]. Previous studies have demonstrated that proper knowledge, attitude and practices among consumers are the key elements to ensure food safety [12]. However, there is currently a dearth of information on consumers’ knowledge, perception and behavior in relation to pest ants in domestic kitchens as little research has been carried out on this topic. The purpose of this study was therefore to (i) provide insight on consumers’ knowledge, perception and behaviors regarding infestation of domestic kitchen by ants; (ii) identify pathogenic microorganisms that are harbored by ants which regularly infest kitchens; and (iii) determine the potential of insects to transfer microorganisms to food. AIMS Microbiology Volume 4, Issue 2, 319–333. 321 2. Materials and methods 2.1. Consumer survey A survey questionnaire was designed to shed light on consumers’ knowledge, perception and behavior regarding pest ants in domestic kitchens. In this investigation, 100 participants were selected randomly to represent Mauritian consumers. The target population for this survey was members of the general public of both genders, from diverse educational and ethnic backgrounds. The participants were contacted personally at their place or workplace and requested to fill in the questionnaire appropriately with answers of their choice. They were also informed that all the information collected during the course of this survey would be kept confidential and strictly used for statistical purpose. The questionnaire contained items pertaining to demographic information such as age, gender and marital status as well as a series of close-ended and open-ended questions assessing the knowledge, perception, behavior and control practices of pest ants in domestic kitchens. 2.2. Determination of microbial carriage of ants Microbiological tests were carried out in order to identify any pathogenic microorganisms found on ants commonly present in domestic kitchens. The method of capturing and microbiologically analyzing ants was adapted from Maximo et al. [14] and Ogba et al. [15] respectively. Briefly, 50-ml centrifuge tubes, each containing 10 g of sugar, were heat-sterilized to act as baits. Ants were collected during the period of September–October 2016 from 5 different domestic kitchens located in the Northern district of Pamplemousses. This region was chosen since the climate is warmer and relatively more humid, rendering it more conducive for ant breeding and house infestation. The tubes were left on kitchen countertops near (i) windowsills, (ii) sinks, (iii) bread-storage areas, (iv) pantries or (v) microwaves overnight for ant collection. These areas were selected because of the higher incidence of ants as reported by survey-participants and the relative ease of capturing them. Two independent trials were carried out totaling 50 samples (5 kitchens × 5 sampling sites × 2 replicates). Once ants were captured, tubes were capped and refrigerated at 4 °C for 1 h to immobilize the insects. These ants were identified taxonomically as Technomyrmex difficilis and Solenopsis geminata by consulting the resource AntWeb v. 7.7.4 (antweb.org) as well as comparison of their key features with a collection of ant specimens from the Zoology Laboratory of the University of Mauritius. Their identity was further confirmed by consulting a local entomologist. For the microbiological analysis of ants, 40 ml of 1.0% sterilized buffered peptone water (Hi-Media) was added to each tube and incubated for 24 h at 35 °C for enrichment. A loopful of the enriched culture was streaked on Violet Red Bile Agar, Eosin Methylene Blue Agar, Slanetz and Bartley Medium, Bacillus Agar, Iron sulphite Agar, McBride Listeria Agar, Xylose Lysine Desoxycholate Agar and Potato Dextrose Agar (PDA) to detect the presence of coliforms, Escherichia coli, Enterococcus faecalis, Bacillus species, Clostridium perfringens, Listeria monocytogenes, Salmonella spp. and yeasts and molds respectively. Plates were incubated at 35 °C for 24 h after which they were observed for presence or absence of growth. AIMS Microbiology Volume 4, Issue 2, 319–333. 322 2.3. Determination of transference rate of tracer bacteria E. coli to food by ants This experiment was carried out to determine whether ants have the potential to transfer pathogenic bacteria from a contaminated source to food. Twenty 50-ml centrifuge tubes were autoclaved, to which sterile sugar (10 g) was subsequently added to serve as an attractant. Sugar from ten of the tubes was inoculated with 200 μL of