{"paper_id":"165c0214-6781-49f1-afa6-742c881be7b9","body_text":"1\n1 Influences on limited antimicrobial use in small-scale \n2 freshwater aquaculture farms in central Thailand\n3\n4 Nour Alhusein1*, Boonrat Chantong2, Sarin Suwanpakdee2, Anuwat Wiratsudakul2, Virginia C. \n5 Gould3,#a, Kantima Wichuwaranan 4, Kornrawan Poonsawad 4, Varapon Montrivade 4, Nutcha \n6 Charoenboon1, Luechai Sringernyuang 4, Matthew B. Avison 5, Helen Lambert 1, Walasinee \n7 Sakcamduang2, Henry Buller6†¶, Kristen K. Reyherg¶*\n8\n9 1 Population Health Sciences, Bristol Medical School, University of Bristol, Bristol, UK\n10 2 Faculty of Veterinary Science, Mahidol University, Nakhon Pathom, Thailand\n11 3 Bristol Veterinary School, University of Bristol, Bristol, UK\n12 4 Faculty of Social Science and Humanities, Mahidol University, Salaya, Nakhon Pathom, \n13 Thailand \n14 5 School of Cellular and Molecular Medicine, University of Bristol, Bristol, United Kingdom \n15 6 College of Life and Environmental Sciences, University of Exeter, Exeter, UK \n16 #aCurrent Address: School of Psychological Science, University of Bristol, Bristol, United \n17 Kingdom\n18 *Corresponding authors:\n19 nour.alhusein@bristol.ac.uk (NA)\n20 Kristen.Reyher@bristol.ac.uk (KR)\n21\n22 ¶HB and KR are Joint Senior Authors\n23 †Deceased\n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \nNOTE: This preprint reports new research that has not been certified by peer review and should not be used to guide clinical practice.\n\n2\n24 Abstract\n25 Recent years have shown substantial growth both in the scale and the spread of freshwater \n26 aquaculture in Thailand, raising concerns about potential widespread antimicrobial use. This \n27 mixed-methods study used surveys and qualitative interviews to examine conditions of \n28 freshwater aquaculture farming in central Thailand in relation to animal health, disease \n29 management and patterns of antimicrobial use. Freshwater aquaculture in this area of \n30 Thailand was largely a domestic venture operated as a source of additional household income \n31 to increase financial security. Aquaculture was often integrated with other types of farming; \n32 initial outlay was reduced by repurposing unused crops, food, or animal manure (e.g. chicken \n33 droppings and pig dung) to fertilise aquaculture ponds. Among farmers representing twenty \n34 farms who were surveyed during 2019, only six farmers said they used antimicrobials. These \n35 included oxytetracycline, enrofloxacin, norfloxacin, ciprofloxacin and sulphonamides. \n36 Farmers doubted the benefits of using antimicrobials to treat aquatic animals; some believed \n37 antimicrobials stunted growth. The high cost of medicines and prohibitive regulations also \n38 discouraged antimicrobial use. Farmers linked disease occurrence to changes in the weather, \n39 the emergence of new diseases and variable water quality. They relied on farm management \n40 practices to maintain the health of their aquatic animals, using lime and salt to maintain and \n41 improve water quality and pH and to disinfect aquaculture pools. Farmers also reported \n42 obtaining juvenile fish and shrimp selectively from farms known to produce healthy stock. \n43 Specialised veterinary services for aquatic farming were rare, so farmers relied on their own \n44 experimentation with medicines, peer advice and recommendations of shopkeepers who sold \n45 both aquatic feed and medicines. This study unexpectedly reveals limited use of \n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n3\n46 antimicrobials linked to socio-economic and ecological features of small-scale family \n47 aquaculture farms.\n48\n49 Introduction\n50 Fast-developing aquaculture sectors in several Southeast Asian countries have been seen as \n51 contributors to the rapidly expanding and diversifying antimicrobial resistance (AMR) \n52 documented in animal production systems over the last two decades [1-3]. Analysis of point \n53 prevalence surveys reporting AMR from aquatic food animals in Asia has found concerning \n54 levels of resistance to medically important antimicrobials in foodborne pathogens [4]. \n55 Increased antimicrobial use (AMU) is associated with the emergence of AMR in bacteria, with \n56 significant implications for both animal and human health [5]. Schar et al. estimated that by \n57 2030, AMU in aquaculture will constitute 5.7% of global AMU and will carry the highest use \n58 intensity per kilogram of biomass [6].\n59\n60 Thailand has been part of the Asian aquaculture surge, being ranked 13th among aquaculture \n61 country producers with a production of 889,891 tonnes of live weight in 2017 [2]. Its \n62 industries predominantly centre on coastal production of shrimp, molluscs and prawn in salt-\n63 water ponds. More recent years, however, have seen the significant growth of inland, fresh-\n64 water aquaculture in Thailand [7]. Here the early emphasis has been rather more on the \n65 production of fish species, notably catfish and tilapia, though shrimp production has grown \n66 rapidly and displayed a similar dynamism to that of the coastal regions [8]. In 2015, freshwater \n67 aquaculture in Thailand accounted for 45% of total aquaculture production value, with the \n68 remainder from marine/coastal aquaculture [7]. \n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n4\n69 The growth of inland Thai aquaculture (both shrimp and fish) has given rise to many  debates, \n70 despite its very clear profitability for those involved [9,10]. There are concerns about the \n71 sustainability of the older model of domestic rice production in many areas of central Thailand \n72 - the widescale and highly profitable conversion of paddy fields to aquaculture ponds has \n73 accompanied this transition, changing the nature and regulation of the rural communities \n74 that rice traditionally sustained [8,11,12]. A second concern has been the use, over-use and \n75 management of fresh-water resources, both river and canals, as critical and replenishable \n76 growing media for aquatic species at a time when other uses of water are being employed, \n77 particularly as a means of moving effluent and wastewater out of sites of production. Little \n78 empirical evidence currently exists as to whether inland aquaculture alone makes a major \n79 contribution to river and canal pollution or whether, alternatively, aquaculture can place itself \n80 principally as the victim of pollution from urban and industrial sources [13].\n81\n82 A third concern about Thai aquaculture, alongside most agriculture systems globally, has been \n83 expressed: that emergence of AMR in environmental or animal-borne pathogens is driven by \n84 AMU on aquaculture farms, resulting in the increased potential for AMR infections in humans \n85 [14-17]. Indeed, the rapid growth and spread of the Thai marine aquaculture industry in the \n86 early 2000s was characterised by high profit margins sustained by a wide range of chemical \n87 inputs (including the use of antimicrobials) to prevent and treat diseases as well as improve \n88 water quality [18]. These inputs took place in an environment where many production-related \n89 diseases were present and unchecked, where increasingly concentrated fish and shrimp \n90 species were vulnerable to disease outbreaks and where, initially, a relatively unregulated \n91 legal framework governed AMU in animal farming [19-21]. The result was that much of the \n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n5\n92 Thai aquaculture sector developed with relatively high levels of prophylactic AMU being the \n93 norm, particularly in the coastal zone [22].\n94\n95 Recent years have seen establishment of several significant regulatory and policy initiatives \n96 in Thailand to survey, monitor and reduce AMU in aquaculture [23,24]. Under these \n97 regulations, only the antibacterials oxytetracycline, tetracycline hydrochloride, \n98 sulfadimethoxine, trimethoprim, sulfadimethoxine/ormetoprim, amoxicillin and enrofloxacin \n99 are authorised in aquaculture [23]. Several market initiatives have followed suit with major \n100 retailers and food processing groups seeking to limit AMU within their Thai supply chains (e.g. \n101 the Raised Without Antibiotics initiative) [25]. The first five-year National Strategic Plan on \n102 AMR in Thailand was published in 2016, covering the period 2017 to 2021 [26]. Since then, \n103 legislation has tightened the availability and accessibility of antimicrobials for aquaculture and \n104 reinforced the role of the veterinarian in antimicrobial prescribing.  Moreover, AMU for \n105 growth promotion has been prohibited, and certain antimicrobials considered critically \n106 important for human health have been made unavailable for use in aquaculture. These \n107 changes were in parallel with those in many other major international food-trading nations.\n108\n109 Investigating current aquaculture practice\n110 Fifty kilometres outside Bangkok, a wide river flows south through the agricultural district of \n111 ‘Plakat’ [a pseudonymised district name has been used to maintain anonymity], part of a \n112 larger administrative province. This is a flat fertile landscape with an elaborate network of \n113 irrigation systems and canals, bearing witness to agricultural and industrial activities following \n114 a period of land reformation. Rice fields and other field crops, vegetable farms, fish ponds, \n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n6\n115 shrimp ponds, chicken, duck and pig farms are numerous in the district. There are also \n116 industrial factories and solar energy power stations. In agricultural terms, over the last two \n117 decades there has been a notable shift (as there has across much of central Thailand) from \n118 the more longstanding production of rice to more profitable, low salinity freshwater fish \n119 (notably tilapia) and shrimp production. Local fish markets serving Bangkok and nearby \n120 provinces provide critical commercial infrastructure, along with a range of aquaculture sector \n121 industries which include food-processing factories as well as feed and animal suppliers. With \n122 such animal production - and its associated commercial and market exigencies - come new \n123 and different pressures notably around health (both animal and public health), disease \n124 management and resource control, which interlink with the growing national agenda of \n125 antimicrobial surveillance and reduction mentioned above.\n126\n127 In an effort to identify the principal drivers of AMU in aquaculture and understand how inland, \n128 small-scale aquaculture farmers respond to shifts in animal health practice and regulation, \n129 our team spent a year in Plakat district, conducting research with freshwater prawn and fish \n130 farmers as part of a wider project to build a holistic picture of AMR drivers in Thailand from \n131 the One Health (human-animal-environment) perspective [27]. We investigated aquaculture \n132 farm management practices and attitudes towards AMU as well as the information and \n133 advisory networks they had for information about antimicrobials and aquatic animal health.\n134\n135\n136 Materials and Methods\n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n7\n137 A mixed methods approach was employed comprising a cross-sectional survey along with \n138 semi-structured interviews. \n139\n140 Materials and participant recruitment\n141 Primary research was undertaken through a questionnaire-based farmer survey \n142 supplemented by additional information obtained by a parallel series of semi-structured local \n143 community (household) interviews, undertaken as part of this research programme but \n144 oriented more specifically to human health and AMU (Fig 1).\n145\n146 Fig 1. A diagram showing the methods and the data sources involved in this study. \n147\n148 Farmer survey\n149 The purpose of the questionnaire survey was to explore the aquaculture farms and animal \n150 health management practices as well as AMU. A questionnaire was developed by the Thai and \n151 UK research teams that combined closed, open-ended and multiple choice questions covering \n152 three main areas: first, farm structure and farmer demographics; second, animal production \n153 and health; and third, medicine use including knowledge, advisory systems and networks \n154 used by farmers, with a particular emphasis on antimicrobials (Farmer survey in S1 Appendix). \n155 Data collection was conducted between October 1, 2019, and ended on December 20, 2019. \n156 A purposive sampling was followed  based on geographical area and farm type. The farms \n157 were geographically centred around the principal settlement of ‘Plakat’ but stretched across \n158 the river basin and extensive local canal system. A total of 20 aquaculture farms were \n159 recruited: nine fish farms and 11 shrimp farms. The survey was conducted face-to-face (social \n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n8\n160 distancing in place due to COVID-19 pandemic) with the farm owner by different members of \n161 the research team (BC, SS and/or AW). On a few occasions, co-workers were present and \n162 contributed to the discussion. Informed consent was obtained from all participants. All \n163 participants needed to sign a consent form before participating in the study. Only adults \n164 participated in the study. Open-ended questions were all in Thai, were audio recorded and \n165 subsequently translated to English and analysed. Responses to the closed and multiple-choice \n166 questions from the questionnaire were analysed and will be reported in future manuscripts. \n167 (BC, SS, AW) are researchers with a speciality and training in veterinary sciences. \n168\n169 Additional interviews with households\n170 Additional qualitative interviews were undertaken with local residents, including aquaculture \n171 farmers, in the district as part of the wider project’s aim to understand local attitudes towards \n172 human health, AMU and AMR. These also provided additional information on aquafarming \n173 and animal health practices. Nineteen households were interviewed by researchers (KW, KP \n174 and NC) and later analysed for the aquafarming theme. Data collection took place between \n175 February 2019 and August 2021. The sample was purposively selected based on geographical \n176 area and socio-economic diversity. Initial contact was made by the village health volunteers \n177 (community health workers) who introduced the study on behalf of the research team [28]. \n178 Households who were happy to participate were introduced to the researchers. All visits were \n179 made on an invitation/permission basis from the key informant of the household. Social \n180 distancing rules were applied due to COVID-19 pandemic.  On average, at least 5 visits were \n181 made to each household. Monthly visits were made to some households with which the \n182 research team had developed deeper relationships. Time per visit varied from the early (30-\n183 60 min) to the later (1-3 hours) period of fieldwork. An informed consent was obtained from \n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n9\n184 all the households. Only adults participated in the study. Other adult family members \n185 contributed to interviews if they were present. Interviewees were in effect self-selected from \n186 among family members according to availability, but efforts were made to ensure a \n187 representative spread of occupations and ages across the overall sample. The three main \n188 themes for investigation included living conditions (livelihoods, food and water sources), \n189 health and treatment-seeking (including medicine use) (Interview topic guide in S2 Appendix). \n190 Researchers used both audio recorders and field notes to record these visits. Out of 19 \n191 households, almost half (nine) were identified as running small aquafarming businesses. (KW, \n192 KP, NC) are trained qualitative researchers with background in anthropology. \n193\n194 Data management and analysis\n195 Interview transcripts and responses to open-ended questions were recorded, transcribed and \n196 translated into English. These transcripts were then coded using open coding and analysed \n197 thematically [29] using NVivo qualitative data analysis software (Version 12, QSR \n198 International, Melbourne, Australia). A thematic analysis approach enables exploring the data \n199 in depth and identifying patterns of meaning across the data.  The first step involved NA \n200 reviewing the transcripts through the stages of familiarization and assigning initial inductive \n201 codes to a sub-sample of transcripts. An initial coding framework was developed and \n202 discussed with the rest of the research team. The remaining transcripts were then indexed by \n203 NA using the coding framework and refinements were made as necessary. NA then re-read, \n204 reviewed and clustered the identified codes to form inductive themes and sub-themes and \n205 assigned names to the themes. The process was iterative and since the data were translated \n206 from Thai, cross-checking of accuracy of interpretation went on throughout the analysis \n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n10\n207 through continuous discussion with the Thai researchers and the rest of the research team. \n208 Illustrative quotes from the farmer survey are presented with descriptors including the farm \n209 type (fish or shrimp) and the farm ID number (e.g. fish farmer F3 or shrimp farmer S4). \n210 Illustrative quotes from household data are presented with descriptors including the farm \n211 type (fish or shrimp) and household ID number (e.g. fish farmer HH38). \n212\n213 Ethics\n214 Formal ethical approval was received from Mahidol University Social Science Institutional \n215 Review Board [(Certificate of Approval No. 2019.026.0702), (MUSSIRB No. 2019/024 (B2)], \n216 and Faculty of Veterinary Science, Mahidol University – Institute Animal Care and Use \n217 Committee (FVS-MU-IACUC), (license no: UI-01292-2558) and registered with University of \n218 Bristol, UK.\n219 The study used mixed methods including a survey of farmers and semi-structured interviews \n220 with adult members of households in the local community. For the farmer survey, written \n221 consent was obtained from all participants. For semi-structured interviews, written consent \n222 was requested before the commencing of data collection from all households. Some \n223 households initially preferred to give a verbal consent. In this case, the verbal consent was \n224 obtained and witnessed by another member of the research team, other household members \n225 and village health volunteers (Community health workers) who helped in the recruitment. \n226 Subsequent confirmation of consent was ensured on an ongoing basis (e.g. repeated requests \n227 for permission for household visits), in line with the Association of Social Anthropologists of \n228 the UK (ASA) Ethical Guidelines for good research practice (ASA 2021). Later, written consent \n229 was obtained from all households for other data collection activities (e.g. household survey, \n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n11\n230 stool samples, reported elsewhere). For each activity, the research team explained to \n231 participants the study objectives and procedures, the voluntary nature of participation, \n232 principles of confidentiality and anonymisation of data. The consenting procedure (written or \n233 verbal) was approved by Mahidol University Social Science Institutional Review Board.\n234\n235\n236 Results\n237 Farm characteristics\n238 Most farmers had members of their families living on the farm and working with animals. All \n239 farms were owned by the farmer; none were contracted from a company. Fifteen farmers \n240 had established the farms by themselves, three were inherited and two rented. Fish farmers \n241 mainly raised tilapia ( Oreochromis niloticus). Other species included giant gourami \n242 (Osphronemus goramy), Jullien’s golden carp (Probarbus jullieni) and other carp species \n243 (Cyprinidae). Shrimp farmers raised Pacific white shrimp ( Litopenaeus vannamei) and giant \n244 freshwater prawn (Macrobrachium rosenbergii) (Farm characteristics in S3 Table). \n245\n246 Household characteristics\n247 It was very common for household members to have multiple occupations (Household \n248 characteristics in S4 Table). Of the nine households, seven had private ponds used to raise \n249 fish or shrimp. Others had a cage (or floating baskets) used for raising fish in shared ponds or \n250 in the river (Fig 2). \n251\n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n12\n252 Fig 2.  Left: Fishpond. Middle: Drained fishpond with protective nets. Right: Shared fishery \n253 in a pond or river.\n254\n255\n256\n257\n258\n259 Three main themes and nine subthemes were generated from the analysis (Fig 3). \n260\n261 Fig 3. Thematic map of the themes and subthemes identified in the data.\n262\n263 Health management practices  \n264 Pond preparation and disinfection\n265 Farmers grew fish and shrimp in ponds dug out of the ground. To prepare the ponds for new \n266 stock, farmers would leave emptied ponds to dry out, clean the floor of the ponds and then \n267 add new water. Drying periods varied; one farmer spoke of allowing the floor to dry for 40-\n268 60 days while others mentioned a few days to two weeks. Once the soil was dry, most farmers \n269 scattered lime (calcium hydroxide or calcium carbonate) on the pond floor to disinfect the \n270 ground. Farmers also reported using other chemicals including herbicides or quicklime \n271 (calcium oxide) with two interviewees speaking of disinfecting their ponds with ‘germ-killing’ \n272 medicines (a local term generally used to refer to antibiotics or disinfectants). Treated ponds \n273 were left between one to 10 days, before water was added.\n274\n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n13\n275 “We put in lime first to kill the germs... \n276 Researcher: For how many days, approximately? \n277 “Three days.” Shrimp farmer S4\n278\n279 “To prepare the ponds, we fill them with water. Fill it up to the height that we think \n280 the fish can go in. Then disinfect the ponds using ‘germ-killing’ medicines.” \n281 Researcher: Do you also use lime before [putting the water in] at all? \n282 “No, no need.” Fish farmer F8\n283\n284 Almost all farms obtained water for the ponds from the canal system connected to the river. \n285 Water levels were monitored and usually adjusted to a level of around one metre depth. \n286 Additives were often added to the ponds. These included salts, micro-organisms, ‘germ-\n287 killing’ medicines and fertilizers. The pond water then was usually left for around a week \n288 before the fish or shrimp were introduced. One farmer mentioned waiting for the water to \n289 ‘turn green’ before adding juvenile shrimp.  Some farmers also checked the pH of the water. \n290\n291 “We scatter microbes [micro-organisms] and germ-killing medicines into the \n292 ponds, then we fill in water. After three days or a week, the shrimplets [baby \n293 shrimp] can go in.” Shrimp farmer S5\n294\n295 “Sometimes we also add salt into some ponds to treat the soil and the water, to \n296 not let the water be too acidic or basic.” Fish farmer F5\n297\n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n14\n298  “[…] add water. followed by tablet fertilizer to prime the floor. That’s it.” Shrimp \n299 farmer S10\n300\n301 “Interviewee: When the water goes in, we add microbes [micro-organisms] to \n302 treat it first. \n303 Researcher: That’s before you put the shrimps in. \n304 Interviewee: Yes. We use lime and microbes.  \n305 Researcher: Do you buy the microbes or do you make that yourself? \n306 Interviewee: We buy. Sometimes we use [product name]. We use [product name] \n307 to ferment first, then we use [another product] to adjust the water condition… \n308 Researcher: From then, do you leave it for many more days? \n309 Interviewee: We leave it for another week before we put the shrimp in.” Shrimp \n310 farmer S4\n311\n312 Stock management \n313 Buying the aquatic animals from trusted sources was vital to farmers as such a practice meant \n314 fewer diseases later. New fish were usually kept together in a single, protective ‘nursery pond’ \n315 for an initial two and a half to three months. When the fish had grown in size and weight, they \n316 were divided between different ponds. Just over half the farmers reported co-housing \n317 different species. It was common for shrimp farmers to raise giant freshwater prawns and \n318 white shrimp together in the same pond. One farmer raised fish and shrimp together. Other \n319 animals like poultry (in small-scale production) roamed freely around the ponds or walked on \n320 the nets covering the ponds. \n321\n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n15\n322 “But if you want to raise the fish together with the shrimp, you have to put the \n323 shrimp in first. We take that water to test in a lab; there’s a lab in [town]. If the \n324 water passes [the standards], then we can release the shrimps. Fifteen days after \n325 the shrimps go in, the fish can go in – tilapias and seven-striped carp and Chinese \n326 fish. Then we raise them.” Fish farmer F7\n327 Feeding and nutrition\n328 Farmers explained that the aquaculture farming rearing cycle was generally operated over a \n329 one-year period. This period could be reduced to six to eight months depending on the \n330 farmer’s feeding style, whether they used organic feed or commercial supplements. \n331 Commercial supplements accelerated the growth of fish and shrimp, but not everyone could \n332 afford them. Some farmers used animal manure, sourced from their other livestock such as \n333 ducks, chicken, or pigs, to feed fish or shrimp in the ponds. This practice was seen as a way to \n334 reduce cost as well as being wise use of resources. One farmer mentioned having direct \n335 pipelines from their pig sheds through which dung was pumped into the ponds. Another \n336 farmer sold the extra dung they had to other farmers. Animal manure usage, however, was \n337 disapproved of by some other farmers due to its unpleasant smell. Farmers also reported \n338 using other types of organic or locally available sources of fertilizers such as vegetables from \n339 their garden, lotus, hay or rice bran either alone or mixed with powdered fishmeal usually \n340 given to small fish. One farmer mentioned using the wastewater from a fertiliser factory and \n341 another fed fermented pineapple peel to the shrimp. One participant described using unused \n342 noodles from a nearby factory. Vitamins were also used, though one farmer described them \n343 as a waste of money.\n344\n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n16\n345 “I took pig dung to feed the fish. When I wash the pig dung from this coop, the \n346 water flows down two pipes; it flows into the fishpond.” Fish farmer HH38\n347\n348 “Feed this fish only grain and rice bran. Mix rice bran with pellet food. Chicken \n349 manure, […] even the low cost, but the son doesn't like it, he doesn't want it. \n350 Chicken manure is smelly too. Big pond over there [ different farm]. The owner of \n351 the big pond raised it with chicken manure. Feed both pellet food and chicken \n352 manure. That fishpond was delivered by a truck of chicken manure. Feeding the \n353 fish like that makes the fish grow fast.” Fish farmer HH03\n354\n355 “The noodles were brought to the fish to eat, but the noodles had to be marinated. \n356 Other farms do this. It has to add molasses water to mix in the tank as well. They \n357 buy from sugar factories.” Fish farmer HH03\n358\n359 “I feed the fish with leftover food, vegetables that I bought to cook and then I throw the \n360 rest to the fish in the pond.” Fish farmer HH25\n361\n362 Disease control and medicine use\n363 Disease drivers\n364 Many farmers reported that diseases amongst their stock had increased over the last decade. \n365 While some farmers offered no explanation for this, others felt different factors were \n366 responsible including weather changes, the emergence of new diseases and decreasing water \n367 quality due to pollution. \n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n17\n368\n369 “In the past they never got sick; if we found a few sick or dead ones we will just \n370 remove them.” Shrimp farmer S11\n371\n372 “Lately, we don’t know what’s wrong with the fish, but they keep dying.”  Fish \n373 farmer F6\n374\n375 “If the water is good and the weather is good, we don’t need to use any medicine.” \n376 Fish farmer F4\n377\n378 There was no consensus whether it was the heat or the cold that caused disease in the fish or \n379 shrimp, but it seemed that the extreme weather conditions led to disease spreading. \n380\n381 “The disease outbreak is usually at the end of winter/beginning [of] summer. \n382 Maybe, because of the heat, not sure, March-April. I give them medication and \n383 add a lot of water. I don’t know, maybe because it is getting warm around that \n384 time of the year.” Fish farmer F6\n385\n386 “Researcher: You mentioned that the animals died a lot during the past year ... \n387 how did it happen? What were the symptoms? \n388 Interviewee (1st male): It is the heat.  \n389 Interviewee (2nd male): The weather is too hot, and they [the fish] got sick.”  Fish \n390 farmer and co-worker F7\n391\n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n18\n392 “If the weather keeps getting colder, we will probably have to buy them \n393 medication.” Fish farmer F9\n394\n395 “They are currently healthy, because right now the temperature is dropping. Once \n396 it got cooler, they are able to survive. But they would also start showing symptoms \n397 if it is getting too cold.” Shrimp farmer S5\n398\n399 Water condition was considered vital in raising healthy aquatic animals and was monitored \n400 continuously, largely through attention to its colour, appearance and pH.   Poor-looking water \n401 was often treated using additives such as salt, micro-organisms and ‘germ-killing’ medicines. \n402 Two farmers spoke of the impact of rain on the water conditions. Others spoke of the \n403 importance of oxygenating the water, sometimes artificially. Critically, these water quality \n404 and environmental factors played a part in determining subsequent antimicrobial and other \n405 medicine use. \n406\n407 “After one month, we add more water and salt. We have to take note of the water \n408 colour. If the colour starts to look bad – if it’s not green or clear – then we have to \n409 add salt or minerals.” Shrimp farmer S9\n410\n411 “It depends on the weather and the water. For example, when we left them out during \n412 the rain, then they would start dying. They are unable to survive under that condition, \n413 because of the dropping pH, and alkaline, the shrimps are unable to adapt.” Shrimp \n414 farmer S5\n415\n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n19\n416 “I think the rain is toxic. I think. My opinion, not sure how true it is.” Fish farmer F8\n417\n418 “It depends on the water. If the water quality is good, it would be acceptable for us not \n419 to add any medicine.” Shrimp farmer S3\n420\n421 Half the farmers reported having significant animal disease outbreaks or mass dying events \n422 recently, with six reporting estimated mortality rates of over 20% - in one case, rising to 80% \n423 of stock. One farmer described disease symptoms in fish as “red belly, disease in the eye, \n424 swollen navel” (Fish farmer F9). Another looked for specific symptoms such as bruises as signs \n425 of disease. For most, shrimp or fish feeding rates and mortality levels served as the basic \n426 indicators of animal population health. One farmer mentioned using a lifting net to examine \n427 the fish closely or shining a light at them at night. \n428\n429 “The amount of death is uncertain because we can see only when the dead fish \n430 are floating.”  Shrimp farmer S2\n431\n432 “When they got sick, when they started to die. Once they died, they will float, and \n433 we just need to determine what kills them. Like, if they have bruises, we would \n434 have a look and decide on bacterial or viral, and then we would treat them \n435 accordingly.” Fish Farmer F6\n436\n437 “Researcher: From your experience, you lift Yo (a tool for trapping fishes) to see \n438 how they are? \n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n20\n439 “Yes. We also can see when the fishes come up to the side of the bank. We shine \n440 the light to see them during the night time… we check whether they (shrimp) have \n441 tight texture or black gum…have foods in their intestines.” Shrimp farmer S7\n442\n443 \"It doesn't work\": AMU on aquaculture farms \n444 Farmers were generally divided as to whether AMU on aquaculture farms had increased or \n445 decreased in the last decade. Those that mentioned AMU decreasing argued that medicine \n446 cost was high and that aquaculture farming simply did not generate enough revenue to cover \n447 this cost. Some doubted the benefits of using antimicrobials in treating aquatic stock, \n448 believing antimicrobials actively stunted growth. For a few respondents, government \n449 regulations along with food chain and market imperatives mitigated against the use of \n450 antimicrobials.\n451\n452 “[AMU] decreased, to decrease the running cost of the farm... Because, antibiotics \n453 are expensive and sometimes ineffective.” Shrimp farmer S9\n454\n455 “Mostly, they [aquaculture farmers in general] don’t use [antimicrobials] \n456 anymore… It doesn’t work.” Fish farmer F4\n457\n458 “However, even medicines did not really help… when we give them medications it \n459 looks like they won’t grow, it looks like the medicines stunted their growth.” Fish \n460 farmer F7\n461\n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n21\n462 “I heard that people say it’s not good if we use too much. If we sell the shrimps and \n463 the merchant/buyer checks and detects the drug use, we won’t be able to sell our \n464 shrimps anymore. That is the reason why I try to limit the usage of medicine with \n465 my shrimps.” Shrimp farmer S3\n466\n467 “For medicine, we may not use it at all because the Department of Fisheries already \n468 told that it will be problem if medicine is detected. We used to face that problem.” \n469 Shrimp farmer S6\n470\n471 Such farmers preferred to use other methods to control disease. These included selecting \n472 reputed aquatic animal providers from which to purchase stock, using herbal medicines and \n473 adding vitamins to enhance growth. As stated above, water quality was seen as an important \n474 factor in keeping the stock healthy, often with additions of salts and lime. In the case of major \n475 disease outbreaks, farmers regularly removed any floating dead fish and shrimp and would \n476 deploy emergency harvesting if needed. Emergency harvests were either sold for sauce \n477 production or became landfill. Infected ponds were then disinfected, and water was pumped \n478 out to prepare for a new batch. \n479\n480 “Mostly, I don’t use [antimicrobials]. Because we focus on the species (of aquatic \n481 animals) that we got from different farms…they are not similar.” Shrimp farmer S6\n482\n483 “Now people mostly use organic methods… we just won’t use it [antimicrobials]. \n484 We will concentrate on natural/organic ways.” Shrimp farmer S4\n485\n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n22\n486 “Yes. I use only herbs I bought from the food shop. I remember that it made from \n487 turmeric, which human also can eat … If vitamins, we use sometimes … When I find \n488 that some ponds grow slowly, I make a decision by myself to use vitamin \n489 supplements.” Shrimp farmer S6\n490\n491 “To avoid using antimicrobials, we prepare the pool and water by adding \n492 microorganisms; oxygenate the water, to help strengthen the animals, so that they \n493 won’t be bothered by any diseases.” Fish farmer F3 \n494\n495 “Researcher: What did you do with the dead ones, did you bury them? Or sell them? \n496 Interviewee: We sold them. We sold them as “Phla-ra” [salt-fermented fish (not \n497 fish sauce) commonly used in many Thai dishes]. Thirteen tons of Phla-ra.” Fish \n498 farmer F8\n499\n500 “During [disease outbreak] we catch them; if we see they die, we must hurry to \n501 catch them. If we let it be, they will all die.” Shrimp farmer S8\n502\n503 Other farms surveyed reported that their use of antimicrobials had increased in recent years \n504 in response to growing disease prevalence. Prophylactic use of antimicrobials was often \n505 described, particularly when an outbreak on a neighbouring farm was discussed. As a \n506 treatment, antimicrobials were largely considered to be a last resort and were far from being \n507 seen as being effective in treating sick animals or populations. Most farmers were unable to \n508 name specific antimicrobial medicines, recognising them rather by their appearance or \n509 characteristics. \n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n23\n510\n511 “I think the use [of antimicrobials] is increased. Nowadays, diseases of aquatic \n512 animals are more and more every day. We need to use antibiotics for curing their \n513 diseases.” Fish farmer F2\n514\n515 “However, I occasionally give them medicine for disease prevention if there is a \n516 disease outbreak and others say I should give them as a prevention measure.” \n517 Shrimp farmer S4\n518\n519 “[…giving them antimicrobials] To stimulate them. […] To prevent them from \n520 getting sick… we don’t give them that often, sometimes we would refrain from \n521 giving them the medication. If they look responsive, we won’t give them any.”  \n522 Shrimp farmer S3\n523\n524 “We don’t know what else we could do. We already try changing the water, and \n525 aerating; we already did everything, but it didn’t work. So, we have to turn to using \n526 medications.” Fish farmer F6\n527\n528 “[…] Like when they ate [the medicine] for seven days in a row, they stop [dying]. \n529 However, if we stop giving them the meds, they will start dying again. They are not \n530 completely cured. We don’t know how to solve this.” Fish farmer F8\n531\n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n24\n532 “If we see giant freshwater prawns die, then we use yellow medicine. That’s it. \n533 Nothing much here because we don’t want to invest too much. The price of the \n534 shrimp [to sell] is low.” Shrimp farmer S3\n535\n536 Out of the 20 farms, only six farms reported using antimicrobials including oxytetracycline, \n537 fluoroquinolones (enrofloxacin, norfloxacin and ciprofloxacin) and sulphonamides. Farmers \n538 reported purchasing antimicrobials from feed companies or stores and mixing these with feed \n539 on the farms. Farmers did not mention obtaining antimicrobials from veterinarians. Other \n540 additives were also used including multivitamins, amino acids and probiotics. The dosage and \n541 frequency were mostly determined by the farmer and varied significantly between farms, \n542 suggesting the absence or lack of knowledge of common guidelines within this small-scale \n543 commercial aquaculture sector. Treatment plans were pond- and population-based. When \n544 some fish or shrimp showed signs of sickness, the entire pond would be treated with the \n545 medicines mixed with the feed or water and spread manually into the pond. \n546\n547 “There is a powder medicine that we can mix in water and put in the pond.” \n548 Researcher: That would be for the entire pond? \n549 “Yes. Some people would mix it in the food. It’s different for each farm.” Shrimp \n550 farmer S11\n551\n552 “We cure them all in a pond. If this pond was cured, I can leave them and take care \n553 of the next pond.” Shrimp farmer S8\n554\n555 Farmers’ perspectives on AMU and AMR  \n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n25\n556 Farmers were asked what they thought about the long-term impact of AMU on humans, \n557 animals or the environment. Misinformation or lack of awareness was evident as some \n558 farmers felt there was no negative impact from AMU.\n559\n560 “I haven’t seen any effects from the use of antimicrobial so far, neither has anyone \n561 come to clarify/educate us about it.” Fish farmer F3\n562\n563 “It’s not harmful to humans…  My opinion would be that our use of medicines isn’t \n564 harmful and doesn’t affect the consumer; even the biologist that gave us the \n565 medication said that there is no effect.” \n566 Researcher: What about these medicines – germ killer? Do you think they have any \n567 effects on the consumer? \n568 “No.” Shrimp farmer S4\n569\n570 “Researcher: Do you know about the effects of using antibacterial drugs on human, \n571 animals and environment? Is there any effect when we use them for a long time?\n572 “Interviewee: No, I don’t know. I don’t really study about this, so I don’t know \n573 much.” Shrimp farmer S7\n574\n575 Other farmers believed that AMU could affect consumers due to chemicals accumulating in \n576 the body and subsequently causing illnesses. However, these farmers explained that using \n577 antimicrobials was necessary, especially in large farms with many animals.\n578\n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n26\n579 “I know that if we use a significant amount of medicines with the animal, it will \n580 affect the consumer as well. The antibiotics we used would accumulate in the \n581 human body, causing it to resist drug action afterwards. So, yes. I know that it \n582 would cause drug resistance in the human body when we use it to treat illness later \n583 on.” Fish farmer F6\n584\n585 “If we use too much medicine for a long time, it would definitely be harmful to humans \n586 because it would accumulate in our body. But it is necessary to use medication because \n587 we need to medicate the animal when they get sick, especially for those big farms that \n588 have a lot of animals.” Shrimp farmer S11\n589\n590 Advice and support networks for disease management and AMU provision\n591 The farmers interviewed stated that they had autonomy over the health management and \n592 treatment decisions taken on their farms. Farmers tended to learn what treatment to use \n593 through their own experimentation with different products and their impact upon illness \n594 symptoms and animal health. \n595\n596 “I decide from the symptom whether to use Oxy [oxytetracycline], Sulfa \n597 [sulfonamides], Enro [enrofloxacin], or others. There are 2 to 3 primary medicines \n598 that I use nowadays.” Fish farmer F8\n599\n600 “If I use any product, and it’s good, then I continue using that [laugh]… I have no \n601 one to do research for me. So, I must do it myself. If I give the fishes any medicines \n602 and see that during the three days they get better, then I continue using that one. \n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n27\n603 But if they don’t get better within three days, then I have to change the medicine.” \n604 Fish farmer F8\n605\n606 Some sought treatment recommendation and advice from more experienced peers, animal \n607 husbandry personnel and from storekeepers who sold aquatic food and medicines. Farmers \n608 occasionally referred to these sellers and personnel as ‘vets’; farmers would describe the \n609 symptoms to the sellers and the sellers dispensed the medicines. Some stores also offered \n610 water testing services.\n611\n612 “I asked the store owner too, and other people who used the same products or had \n613 the same experiences. So, they can give me advice on what to do or what medicines \n614 to use.” Fish farmer F3\n615\n616 “I would consult with people at the drug store. They usually have a veterinarian \n617 there. Sometimes I would ask the veterinarians who can prescribe medicine that I \n618 know. I would tell them the symptoms - such as having red scales, red gill and \n619 droopy eyes - then ask which medicine I should use. They would give suggestions \n620 based on that. We don’t meet face-to-face for the veterinarian to check the water \n621 condition and tell us exactly which medicine we need to use specifically for such a \n622 disease. I would be more sure like 100% if that is the case.” Fish farmer F8\n623\n624 Dedicated aquatic veterinary support was considered very rare in the area and farmers took \n625 every opportunity to contact a veterinarian whenever one came into the neighbourhood, \n626 though none of the farmers interviewed reported a veterinarian visiting their farms for routine \n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n28\n627 checks or in case of an illness or outbreak. One farmer said he had attended a training course \n628 on aquatic farming, and another said they had done their own reading. \n629\n630 “At the beginning, I usually ask the agriculturist/academic. But after a while, I \n631 started to study things myself. I tried different things while seeing what other \n632 people use. That’s how I ended up with this product. Actually, others suggest me \n633 to use Amoxy [amoxycillin] as well.” Fish farmer F6\n634\n635 “A fisheries scientist used to come to the district office, so I memorized the names \n636 of the medications. I would call and asked him, whether this meds would work or \n637 not, and he said that it can. So, I made the decision.” Fish farmer F6\n638\n639 “I attended training about fish diseases before. The instructor instructed us to use \n640 which medicine with what symptom. I had the instructor’s phone number, so I used \n641 to call him and ask him for the suggestion of medication relating to the fishes’ \n642 symptom, but I lost the number.” Fish farmer F6\n643\n644\n645 Socioecological influences in the development of aquaculture \n646 Shifting land use from traditional rice farming to other agro-ventures including \n647 aquafarming\n648 Many participants described how land was, in the past, mainly used for rice farming and that \n649 rice fields spread as far as the eye could see. Participants mentioned that, during the 1990s, \n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n29\n650 the use of land started to shift due to a fall in rice prices and subsequent economic loss. \n651 Additionally, participants explained that the rice farming workforce has reduced over the \n652 years due to aging with younger generations not wanting to undertake low-income \n653 occupations. Subsequently, new business ventures started to appear. Factories were built and \n654 other agricultural systems were promoted including gardening, plantations, aquaculture \n655 farming and livestock farming to generate more income.\n656\n657 “Many people are doing the shrimp pond business because the income is good. \n658 Those who cannot do the rice field will have shrimp ponds. The shrimp pond \n659 business has been around for about 10 years because people had left the area and \n660 when they came back, they started a shrimp pond business.” Fish farmer HH25\n661\n662 “Our family switched to raising fish in the old fields, digging old fields into fish \n663 ponds. At that time, farming was not good. Oh, now farming requires a lot of \n664 investment, a lot of things. My parents are old and I can't farm myself. So our \n665 family has switched to raising fish instead.” Fish farmer HH25\n666\n667\n668 “My family used to farm rice. I don't plant rice anymore, now I raise shrimp. In the \n669 past, most of the rice fields were cultivated in this area, 80-90% of the area was \n670 rice fields. Today it is transformed into a pond and mixed farming. It has changed \n671 in the past 10 years from rice fields to shrimp ponds. However, when shrimp prices \n672 are not good, there are farmers who turn back to rice farming. But my family has \n673 always been raising shrimp since we have been raising shrimp and never changed. \n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n30\n674 I've raised white shrimp before. I planted Manila tamarind on the edge of the \n675 shrimp pond, cultivated as an additional income. One year it can sell for over a \n676 hundred thousand baht.” Shrimp farmer HH19\n677\n678 Economic fragility \n679 All the households involved in aquaculture farming also had other sources of income. \n680 Household members worked in various agriculture projects such as rice farming, growing \n681 vegetables, fruit gardens, lotus ponds and aquaculture farming as well as livestock farming \n682 including poultry and pigs. Other occupations such as working in factories and cookery were \n683 also common. Participants explained that having different occupations provided more \n684 financial stability to support families. For example, one household had to close their chicken \n685 farm in 2006 due to avian influenza. However, they had other sources of income from raising \n686 shrimp and growing rice as well as working as a truck driver. Another household, which had a \n687 chicken farm contracted to sell eggs to a larger farm, closed the chicken egg business due to \n688 lower egg prices being imposed by the larger farm. They had to sell all the chickens back to \n689 the larger farm (from which they were originally purchased) at a financial loss. This household, \n690 however, also had a tilapia pond they could rely on. One participant explained that the \n691 concept of holding several farming projects was encouraged by the government-promoted \n692 “sufficient economy” ideology which advocated the wise use of one’s own resources and the \n693 redeployment rather than wasting of these resources.  \n694\n695 “The people choose to do many kinds of agriculture because they might face a lack \n696 of income if one of their businesses has some complications.” Shrimp farmer HH19\n697\n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n31\n698 “My job is to raise fish and plant lotuses. My husband [HH227(02)] is the one who \n699 picked up the lotus from the pond. I also help my husband. My other job is raising \n700 my grandchildren… my husband will go out to feed the fish at 6 am. The pond has \n701 tilapia and yisok [Julien’s golden carp]. Our ponds often sell fish to fishmongers \n702 who sell fish in the fish market. They will come and catch fish at the pond \n703 themselves.” Fish farmer HH27 \n704\n705 “I repaired the equipment myself and also repaired the machines for other people \n706 in the community.” Fish farmer HH27\n707\n708 “My house is sectioned according to the sufficiency economy’s prototype that is \n709 the two rai areas [about 3800-6400 square meters] of the land is for the house, \n710 various kinds of vegetable gardens, a fishpond and a pig shed. The rest of the land, \n711 about 36 rai, is paddy field which is shared with other types of agriculture.” Fish \n712 farmer HH38\n713\n714 Notably, most households were multi-generation residencies where grandparents, siblings \n715 and grandchildren lived together in one house or adjacent houses on the same piece of land. \n716 This concentration of family members provided labour for these multiple occupations. \n717 Participants described helping each other in running different projects with stay-at-home \n718 members (elderly, unemployed) generally helping with commercial cooking (restaurants, \n719 catering) and aquaculture farming or livestock feeding. \n720\n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n32\n721 “This fishpond belongs to my son. I couldn't do it, I let the kids feed the fish. When \n722 catching fish for sale, there will be many relatives who are selling fish in the fish \n723 market... We don't have to do anything. My son works at the university. He \n724 returned on Saturday and Sunday. My daughter is a fish feeder… This pond is \n725 raising tilapia, carp and yisok. It has been raised for 7-8 months.” Fish farmer HH03\n726\n727\n728\n729 Discussion\n730 This study aimed to investigate the socio-economic and ecological conditions of freshwater \n731 aquaculture farming and the drivers for AMU in a district in central Thailand. Our findings \n732 demonstrated that there was limited use of antimicrobials across the farmers in this study.  \n733 Those farmers expressed doubt about the benefits of antimicrobials, both in relation to \n734 disease treatment and aquatic animal growth. High cost and prohibitive market regulations \n735 also discouraged farmers from using antimicrobials. Disease drivers were often linked to \n736 weather changes, the emergence of new diseases and decreasing water quality due to \n737 pollution. Farmers depended on improved farm management practices to maintain the health \n738 of the aquatic animals; when that failed, many farmers applied emergency harvesting \n739 techniques (i.e. no treatment but catching animals that were still alive). Aquaculture \n740 veterinary support was limited in the area and many farmers relied on their social networks \n741 and on experimenting with medicines in relation to decision making about disease treatment. \n742 Aquaculture farms in this area took the shape of domestic ventures operated by family \n743 members. These families were often found to invest in different juxtaposed small businesses \n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n33\n744 in an attempt to maintain or improve the financial stability of the household. Aquaculture \n745 farming was considered a profitable business that did not require high input and labour. Initial \n746 outlay was often reduced by repurposing unused crops, food or animal manure to fertilise the \n747 ponds and stimulate aquatic animal growth instead of, or in addition to, using commercial \n748 feed.  \n749 It is interesting that many farmers in this study stated that they did not use antimicrobials in \n750 their farms. Farmers referred to antimicrobial treatments as ineffective and expensive. \n751 Traditional antimicrobial administration methods are often problematic and complex in \n752 aquaculture; dispensing antimicrobials directly into water or adding antimicrobials to aquatic \n753 feed can be ineffective, possibly harmful and could lead to AMR [30-33]. This is due to \n754 difficulties in adjusting a therapeutic dose uniformly across the pond, the effects of the \n755 natural environment on the antimicrobial as well as associated toxicity (e.g. to denitrifying \n756 bacteria leading to a build-up of toxic ammonia) [31]. Additionally, diseased fish often do not \n757 eat, which reduces the success rate of medicating the feed [31]. Accordingly, specialist advice \n758 is important to facilitate effective antimicrobial administration in aquaculture systems. It is \n759 possible, therefore, that, in our study area, due to the lack of veterinary support, our \n760 participants’ own attempts at antimicrobial treatment failed. This may have resulted in their \n761 opinions about the lack of effectiveness of these drugs, which led them to stop using \n762 antimicrobials and look for alternative disease management options. However, a few \n763 participants still reported using antimicrobials despite limited observed benefit. Other studies \n764 have noted that antimicrobial use may fluctuate from year to year even in the same area, \n765 subject to changing climate and disease incidence; one review article noted that farmer \n766 surveys often described inconsistent reports of antimicrobial use [34]. Another study in \n767 Vietnam that monitored antimicrobial use found that although 45% of farmers believed \n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n34\n768 antimicrobials had no effect on curing diseases, 86% of the farmers who held that opinion still \n769 used antibacterials for either treatment or prophylaxis [35]. These reports suggest that our \n770 results indicating low use of antimicrobials should be interpreted with caution; although the \n771 findings indicate little use, it would be valuable in our study area to see whether abstinence \n772 from AMU continues into the future. Another relevant issue to AMR is the reported use of \n773 fluoroquinolones in this study. One attraction of using fluoroquinolones in aquaculture is that \n774 these molecules are stable in aquatic environments, rendering them easy to manage for \n775 aquaculture disease treatment [36]. Indeed, enrofloxacin is widely used globally [24].  \n776 However, the use of fluoroquinolones in aquaculture has been banned in many countries due \n777 to their importance of some fluoroquinolones to human medicine (especially ciprofloxacin \n778 and norfloxacin). This is because when resistance to one fluoroquinolone (whether used only \n779 in farming, or used to treat animals and humans) emerges, it usually confers resistance to all \n780 fluoroquinolones [15,24]. \n781\n782 Limited AMU in aquaculture, as was observed in this study, would broadly be considered \n783 positive in relation to AMR; to ensure its sustainability, however, improving farm \n784 management practice is also vital to compensate for the health needs of the aquatic animals \n785 as well as to ensure farm biosecurity. Farmers in this study adopted several management \n786 practices they considered important to protect their stock from diseases: pond preparation \n787 and disinfection, healthy stock acquisition and good nutrition regimes, to name a few. These \n788 farms resembled the description of the small-scale aquaculture farms described by the work \n789 of Little et al. [10,37] which describes aquaculture as an activity practiced predominantly by \n790 farmers “for whom aquaculture constitutes one element of a larger total livelihood portfolio”. \n791 In these systems, farmers often adopt lower-input and lower-risk practices similar to \n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n35\n792 extensive farming systems. This is, for instance, by crop diversification, improving pond \n793 preparation, choosing better-quality stock and using improved quality feed regimes, \n794 subsequently reducing disease incidence and the need for treatment [38,39]. Similar multi-\n795 culture agricultural systems such as home garden systems in Vietnam, Indonesia and Sri Lanka \n796 have been encouraged for their socio-economic and ecological benefits, and have been \n797 highlighted for their potential role in alleviating poverty [40-43]. In Thailand, the effects of \n798 the extensification and the diversification of crops, as seen in our study area, has been shown \n799 to lead to resilience in facing diseases (e.g. rotating rice and shrimp production showed \n800 reduced disease susceptibility when compared to back-to-back shrimp farming) [13]. \n801 Governmental support has also provided knowledge and production management plans \n802 which have, in turn, encouraged farmers to establish mixed cropping agriculture [18]. Land \n803 has often been divided into an area for the main farming activity, the secondary activity and \n804 a supplementary activity, and has included plans for production of different types of plants, \n805 livestock and fisheries [18]. Repurposing local resources such as we saw in in our study area \n806 (e.g. using left-over crops and food to feed aquatic animals) has also been encouraged. \n807 Another characteristic of Thai aquaculture farming was that farmers were seen to be well \n808 connected to vertical and horizontal knowledge networks [44]. Their vertical network went \n809 along the supply chain and also included government representatives, pharmaceutical and \n810 feed companies. Farmers also belonged to social networks of interlinked farmer groups and \n811 small clubs known locally as “Chum Rom”. These networks created a rural seminar culture \n812 where farmers shared their knowledge and practices, and industry and government \n813 representatives attended and added to discussions.  \n814\n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n36\n815 Despite improved management practices, findings of our study indicated that farmers were \n816 still facing high disease and mortality rates among their aquatic animals. This could risk the \n817 successful adaptation of extensive farming styles with low AMU, resulting in farmers \n818 switching to more intensified farming practices with higher AMU [36,45]. Reasons for high \n819 disease incidence were unclear but farmers cited water pollution as one of the drivers of \n820 disease occurrence. The effects of water pollution on aquaculture farming have previously \n821 been investigated in Thailand costal aquaculture regions [46,47]. These studies found that \n822 aquaculture water quality was indeed affected by urban pollution in the canals and included \n823 faecal coliforms, human Escherichia coli, tetracycline resistance genes and nitrogen [46]. The \n824 aquaculture water was a source of salinity and herbicides. Additionally, high AMR prevalence \n825 was found to be associated with a high prevalence of faecal indicator bacteria which was \n826 highest in peri-urban canal water feeding the aquaculture systems [47]. Another study in \n827 Taiwan investigated the potential cross-contamination problems between aquaculture \n828 systems and surrounding waters [48]. This work demonstrated that aquaculture activities (i.e. \n829 usage of antimicrobials) impacted the surrounding aquatic environments and, at the same \n830 time, the surrounding anthropogenic activities impacted aquaculture waters. These studies \n831 show that aquaculture (even with low AMU) would still be in danger of transmission of AMR \n832 from the environment. Another factor potentially contributing to environmental AMR \n833 transmission and selection in aquaculture systems is the use of animal manure in fertilising \n834 the ponds [34,36], a practice applied by some of the farmers in our study.  A few of these \n835 even had integrated systems where manure was pumped from other animal sheds (e.g. pig \n836 or chicken) to the ponds. This practice was seen by farmers as economically sustainable. \n837 However, it is well established that manure is a reservoir of resistant bacteria and \n838 antimicrobial compounds [49]. For example, in a study investigating the impact of integrated \n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n37\n839 fish farming (a practice combining livestock and fish farming, where animal manure is shed \n840 directly into fish ponds) on AMR, the level of resistance in Acinetobacter spp. was found to \n841 increase from 1-5% of bacterial isolates resistant prior to integration to 100% resistance to \n842 oxytetracycline and sulfamethoxazole and to more than 80% resistance to ciprofloxacin after \n843 two months [50]. Another study has shown the correlation between diffusion of \n844 fluoroquinolone resistance genes and biofertilizers utilisation in Chinese shrimp aquaculture \n845 [51]. There is certainly a need to raising awareness among aquaculture farmers of the risks of \n846 using contaminated manure as well as the wider role of veterinary and husbandry support. \n847\n848 Another relevant finding linked to drivers of AMU was that participants linked disease \n849 incidence to changes in climate, and particularly the effect of heat on fish health. Prophylactic \n850 AMU was seen more commonly when farmers anticipated weather changes. Interestingly, \n851 increased local temperature has been associated with increasing AMR in human infections \n852 and this association was consistent across most classes of antibacterials and pathogen [52]. \n853 Another study has found that increased temperature was associated with increased odds that \n854 faecal samples from the environment were positive for resistant E. coli [53]. If AMR were also \n855 to increase on fish farms as a function of temperature, it would be expected that \n856 antimicrobials would then have reduced ability to cure bacterial infections in treated animals. \n857 Increased temperature also affects the chemical activity and uptake of medicines in the pond \n858 environment [30]. All these factors mean a possible increase in disease incidence on \n859 aquaculture farms as the temperature rises, along with forward pressure on AMU, further \n860 exacerbating AMR. There is a specific need, therefore, to support farmers in developing \n861 disease management plans as the climate warms. \n862\n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n38\n863 This study was a qualitative investigation, and the findings are specific to the research \n864 geographical area and may not be generalisable to other geographical areas. Future studies \n865 could include several areas with freshwater aquaculture. Farmers accounts of their use of \n866 antimicrobials, disease incidence and mortality rates are subject to recall bias. \n867\n868 Conclusions\n869 Farmers in this study reported limited AMU in small-scale family aquaculture farming due to \n870 intentional and unintentional socio-economic and ecological factors. Governmental support \n871 to encourage reduced disease risk and crop diversification as well as market regulation \n872 concerning the residues of antimicrobials allowed in food may have encouraged farmers to \n873 reduce their use of antimicrobials.  Past experience of ineffective treatment of farmed aquatic \n874 animals with antimicrobials and the high cost of these medicines have also played a part in \n875 the low AMU reported. Increased disease rates were attributed to weather changes, the \n876 emergence of new diseases and decreasing water quality due to pollution. The lack of \n877 specialist aquatic veterinary support might leave farmers subject to pressure from \n878 commercial drug sellers when they seek disease management advice and misuse may also \n879 make treatment failure more likely, particularly as temperatures rise. There is a need to \n880 investigate the effects of climate change on aquaculture farming across Southeast Asia as well \n881 as its associated disease and treatments patterns. Further attention is needed to understand \n882 and raise awareness about the risks of using contaminated animal manure in aquaculture \n883 farming. Future policies should attempt to fill the gap in specialist veterinary provision to \n884 freshwater aquaculture, address the need to provide evidence-based information and advice \n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n39\n885 on farm management practices that reduce the need for prophylactic and therapeutic AMU, \n886 and particularly address disease prevention in the face of a changing climate. \n887\n888 Acknowledgements\n889 We thank all the community members and the farmers who participated in this study. \n890\n . 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MacFadden DR, McGough SF, Fisman D, Santillana M, Brownstein JS. Antibiotic resistance \n1096 increases with local temperature. Nat Clim Chang. 2018;8:510-514. doi: 10.1038/s41558-\n1097 018-0161-6.\n1098\n1099 53. Schubert H, Morley K, Puddy EF, Arbon R, Findlay J, Mounsey O, Gould VC, Vass L, Evans M, \n1100 Rees GM, Barrett DC, Turner KM, Cogan TA, Avison MB, Reyher KK. Reduced Antibacterial \n1101 Drug Resistance and blaCTX-M β-Lactamase  Gene Carriage in Cattle-Associated Escherichia \n1102 coli at Low Temperatures, at Sites Dominated by Older Animals, and on Pastureland: \n1103 Implications for Surveillance. Appl Environ Microbiol. 2021;87:e01468-20. doi: \n1104 10.1128/AEM.01468-20.\n1105\n1106\n1107\n1108 S1 Appendix. Farmer survey\n1109 S2 Appendix. Interview topic guide\n1110 S3 Table. Farm characteristics\n1111 S4 Table. Household characteristics\n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint \n\n . CC-BY 4.0 International licenseIt is made available under a \nperpetuity. \n is the author/funder, who has granted medRxiv a license to display the preprint in(which was not certified by peer review)preprint \nThe copyright holder for thisthis version posted February 13, 2024. ; https://doi.org/10.1101/2024.02.11.24302655doi: medRxiv preprint","source_license":"CC-BY-4.0","license_restricted":false}