1. INTRODUCTION:

For centuries, infectious diseases like tuberculosis, the plague, and others caused immense misery and deaths, with many people dying for lack of access to adequate treatment. Not until the 1920s, to be exact, did the discovery of penicillin represent a turning point in the history of medicine¹. Alexander Fleming discovered penicillin, a naturally occurring antibiotic that was the first to successfully cure bacterial infections and saved countless lives. Due to this discovery, more antibiotics were developed quickly, and by the middle of the 20th century, they had completely changed the way diseases were treated and drastically decreased death rates.

Antibiotics have been used in agriculture and livestock production in addition to saving human lives. Antibiotics were widely employed in animal feed by the 1940s to boost feed efficiency, encourage growth, and stop disease outbreaks in pigs, cattle, and poultry. In large-scale industrial farming, where animals were frequently housed in cramped quarters and at high densities, rendering them vulnerable to illnesses, this technique was very helpful. Farmers could boost output and lower the need for more costly veterinary care by using antibiotics.

This widespread use of antibiotics in animal production also contributed to the global rise in antibiotic resistance, but the environmental impact of antibiotics, particularly their entry into the ecosystem, has not been adequately studied or understood until more recently. As most antibiotics are not fully absorbed by animals, a significant portion is excreted through urine and feces, which are often spread on fields as manure or directly into water systems. As a result, these antibiotics find their way into soil and water bodies, posing environmental and public health risks.

The issue of antibiotics in animal excrement contaminating the environment is becoming more widely acknowledged. Antibiotic resistance in environmental bacteria may result from the introduction of these drugs into soil and water systems. The very medications that were once used to suppress germs can develop resistance to them over time in both soil and aquatic environments. This may lead to the emergence of resistant strains that are challenging to treat with traditional antibiotics, which could result in public health emergencies akin to those that occurred before to the invention of antibiotics.

Antibiotics² in water systems have the potential to upset the equilibrium of microbial populations in rivers, lakes, and seas, thereby affecting aquatic ecosystems. Wider ecological ramifications of this pollution of the environment may include changes to fish populations, nutrient cycles, and even drinking water quality. Antibiotics, for instance, are frequently used in aquaculture to manage infections in fish and other aquatic creatures as well as to encourage growth. These operations' discharge has the potential to contaminate surrounding rivers with high amounts of antibiotics, hence causing ongoing environmental damage.

The issue of environmental antibiotic contamination poses a significant challenge to both public health and environmental protection. The problem lies not only in the overuse of antibiotics in animal agriculture but also in the lack of regulation and the difficulty in monitoring the full extent of their presence in the environment. Because antibiotics often exist in trace amounts in water and soil, quantifying their effects on human health and the environment is complex. Testing and tracking antibiotic levels in different ecosystems require extensive and sophisticated methods that are often not applied systematically.

The intricacy of antibiotic resistance contributes to the challenge of measuring these risks. Resistance may develop as a result of antibiotic usage, abuse, or even low environmental concentrations of the drug. Antibiotics have the potential to encourage the development of resistant bacteria, which can then spread by direct contact with animals or habitats, polluted water sources, or the food chain. This makes it difficult to monitor the transmission of antibiotic-resistant genes and the precise environmental factors that fuel their emergence.

2. SOURCES OF ANTIBIOTIC ACCUMULATION IN ENVIRONMENT:

2.1. Industries:

Research³ conducted on the wastewater of a treatment plant near Hyderabad, India, that supplies bulk pharmaceuticals producers, revealed ciprofloxacin concentrations higher than those considered therapeutic for human plasma. The discharge from pharmaceutical enterprises during the synthesis of antibiotics is a major source of pollution in the environment that affects ecosystems, animals, and people. The staff's level of knowledge and the industry's production capacity determine the volume and concentration of antibiotic waste. Because industrial antibiotic pollution frequently exceeds that from other causes, there are grave implications for the environment and public health.

2.2. Sewage:

Parent compounds and metabolites of antibiotics are excreted into the aquatic environment, with feces and urine accounting for 50–80% of the parent chemical excretion. Surface waterways are exposed to these antibiotics because they are frequently left behind after sewage treatment. Antibiotic pollution in water systems can be further exacerbated by untreated wastewater combining with rainfall or by unintentional leaks from industrial or sewage pipes.

2.3. Land application of municipal waste:

Antibiotic contamination in soils, groundwater, subsurface drainage, and surface runoff is facilitated by biosolids. When medications are disposed of improperly—for example, by flushing them down the toilet or placing them in the trash—they build up in landfills and wastewater treatment facilities (WWTPs), where they become a major cause of local contamination.

2.4. Poultry and livestock:

The consumption of antibiotics by livestock and poultry has significantly increased compared to human antibiotic consumption for medicinal purposes. Indirect antibiotic consumption via food products like meat, and milk, have increased rather than direct consumption.

2.5. Human consumption:

Antibiotic waste can come from both domestic and human sources, such as personal usage by individuals or caregivers, as well as veterinary treatments used as growth boosters. Antibiotics are controlled in developed countries, but they are typically readily available, even over-the-counter, in underdeveloped ones. Unintentional ingestion, spills, and accidental poisoning can result from the improper disposal or storage of leftover antibiotics in homes, especially in those with small children or elderly residents. The aforementioned behavior is indicative of uncontrolled medicine access, noncompliance with prescriptions by patients, adverse drug reactions, and the emergence of antibiotic-resistant microbes that affect aquatic and human environments. Antibiotics are a common unneeded drug (18%) that is disposed of in the trash by 63% of homes in the United States.

3. ANTIBIOTIC ACCUMULATION AREAS IN ENVIRONMENT:

3.1. Occurrence in wastewater treatment plant:

Antibiotics are processed⁴ in sewage treatment facilities (WWTPs) as the last resort before they are released into the environment, but because of their architecture, antibiotics can accumulate there. Studies conducted in the USA, Canada, China, Spain, Sweden, and other nations have revealed a notable accumulation of antibiotics in sewage sludge. For example, substantial quantities of quinolones, sulfonamides, and macrolides were discovered in a Beijing investigation. Year-long research conducted in South China revealed that antibiotic clearance rates in WWTPs ranged from 21% to 100%.

3.2. Occurrence in water:

Antibiotic contamination is increasingly affecting natural water sources, including drinking water, rivers, streams, lakes, seawater, and groundwater.

3.2.1. Drinking Water:

Even tap water, once considered safe, is contaminated with antibiotics such as macrolides, erythromycin, and clarithromycin, as seen in Madrid, Spain.

3.2.2. Rivers, Streams, and Lakes:

In India and Sri Lanka, E. coli strains in rivers like the Brahmaputra and Kelani River show significant antibiotic resistance, especially to Levofloxacin⁵, Ciprofloxacin, and Streptomycin, with municipal canals displaying higher resistance due to lower flow rates.

3.2.3. Seawater:

A study of seawater and farmed prawns in southern India found detectable levels of sulphonamides in farmed prawns, exceeding the maximum residue limits set by European standards, while no antibiotics were detected in wild prawns.

3.3.4. Groundwater:

Groundwater, a primary source of drinking water, shows widespread antibiotic resistance, with four-fifths of global samples containing antimicrobial-resistant bacteria (ARBs). However, little is known about the sources and pathways of their proliferation.

3.3. Occurrence in soil and sediments:

The two primary dumpsites in Chennai are Kodungaiyur, which is 10 km north of the city center, and Perungudi, which is 10 km south. Every day, 5400 tonnes of solid garbage are delivered to these locations, 10,000kg of which are biological waste from 730 hospitals. Pollutants such as plasticizers⁶, bisphenol A, polycyclic aromatic hydrocarbons, polychlorinated biphenyls, dioxins, and furans have been linked to open burning at these dumpsites. Since pharmaceutically active chemicals like fluoroquinolones (FQs) tend to attach strongly to dumpsite soil and sludge, it is imperative to explore their existence due to a dearth of data.

3.4. Occurrence in plant and aquatic animals:

In Kerala, India, Madangchanok et al. found 128 antibiotic-resistant bacteria in mangrove sediments, of which 66% exhibited multidrug resistance (MDR). The production of biofilms was closely associated with the resistance. The bacteria were mostly from 10 taxa, chiefly Bacillus, according to molecular analysis. Antibiotics such as ampicillin, gentamicin, ciprofloxacin, and methicillin could not kill these multidrug-resistant bacteria; certain strains are used as probiotics, plant growth regulators, or food and human pathogens. This resistance highlights the necessity for international regulatory actions to combat antibiotic resistance by posing a major threat to mangrove cattle and public health.

4. ISSUES RELATED TO PRESENCE OF ANTIBIOTIC IN ENVIRONMENT:

4.1. Development of Antibiotic Resistance:

Resistance can develop through two main processes⁷: primary resistance (vertical transmission within the same species) and secondary resistance (from exposure to antibiotics or microorganisms).

4.2. Increased Infections and Medication Failures:

Improper antibiotic disposal causes environmental contamination, leading to the spread of resistant bacteria. This can result in increased infections and difficulties in managing diseases, as antibiotics become ineffective against resistant strains.

4.3. Prolonged Illness and Hospitalization:

Antibiotic-resistant infections require stronger, more expensive antibiotics and longer hospital stays, which burden healthcare systems and increase risks of complications, disability, or death. A Lancet study predicts that AMR could cause 10 million deaths annually by 2050 if unaddressed.

4.4. Diminished Efficiency of Medical Techniques:

Surgical procedures, organ transplants, and cancer treatments rely on effective antibiotics. Rising resistance could jeopardize these life-saving treatments, exposing patients to greater risk.

4.5. Influence on Susceptible Populations:

Vulnerable groups such as the elderly, infants⁸, pregnant women, and immunocompromised individuals are more at risk from antibiotic-resistant diseases, which can lead to severe morbidity and mortality.

4.6. Universal Health Risk:

Antibiotic resistance is a global concern, as resistant bacteria can spread through water, food, and travel. The World Bank warns that AMR could push 28.3 million people into extreme poverty by 2050.

4.7. Restricted Medication Alternatives:

The slow development of new antibiotics limits treatment options. As resistance increases, doctors have fewer effective medications, emphasizing the need for joint efforts from governments, healthcare, and industry to manage antibiotic waste and promote responsible use.

4.8. Increased Healthcare Expenses:

Treating antibiotic-resistant diseases⁹ is more costly than treating infections with sensitive bacteria, due to the need for stronger drugs and longer hospital stays. The CDDEP estimates AMR could cost the global economy up to 100 trillion USD by 2050 if not properly managed.

5. DETECTION METHODS:

5.1. Detection Methods Based on Liquid Chromatography:

Because it can handle materials with low volatility and thermal stability, liquid chromatography is frequently used to find antibiotics in a variety of media, including water, sediment, and food. With¹⁰ its ability to provide both qualitative and quantitative data, high-performance liquid chromatography (HPLC) is widely employed. The ability to meet varied requirements for detection precision and speed of analysis make combined chromatographic techniques with different detectors—like UV spectroscopy, fluorescence detection (FD), and tandem mass spectrometry (MS/MS)—preferred.

5.2. Liquid Chromatography-Ultraviolet:

Water pollution is typically¹¹ detected using UV-visible detectors in conjunction with high-performance liquid chromatography (HPLC). With MoS2-graphene oxide magnetic nanoparticles as adsorbents, Xiao et al. were able to achieve detection limits of 0.25–0.50ng·mL−1 when evaluating antibiotics such as gatifloxacin, pazufloxacin, and levofloxacin. Using this approach, they were able to achieve recoveries ranging from 85.6% to 106.1% with a relative standard deviation of under 9.5%. Similar to this, Turiel et al. determined actual ambient amounts of these antibiotics in water by analyzing quinolone and fluoroquinolone residues in surface water using HPLC with tandem UV detection. They achieved detection limits of 8–20ng/L.

5.3. Liquid Chromatography-Fluorescence:

Despite their widespread use, ultraviolet detectors sensitivity to particular analytes is restricted. Liquid chromatography with fluorescence detection (LC-FLD) is used to achieve greater sensitivity, particularly for analytes with low concentrations. Golet et al. devised an LC-FLD technique¹² to identify fluoroquinolones (FQs) in urban wastewater, effectively measuring nine FQs as well as quinolone pipemidic acid. Yang and colleagues developed an ultraperformance liquid chromatography-fluorescence technique in 2018 that achieves highly precise detection limits of 3.1–11.2ng/L for sulfonamides in water. Yiruhan et al. found extensive contamination in tap water from Guangzhou and Macao by using HPLC to detect fluoroquinolones; detection limits ranged from 1.0–679.7ng/L in Guangzhou to 2.0–37.0ng/L in Macao.

5.4. Liquid Chromatography-Mass Spectrometry:

Because of its great sensitivity, precision, and selectivity, liquid chromatography-tandem mass spectrometry (LC-MS/MS) is frequently employed for the identification of antibiotics. Liang et al. devised a technique that combines liquid microextraction with solid-phase extraction to detect 10 antibiotics in ambient water, with good recovery rates (64.16%–99.80%) and detection limits (LOD) below 1.67ng/mL. Numerous water samples have been successfully treated using this technology. Goessens et al. (2020) achieved good linearity (R² ≥ 0.993) and detection limits between 10–50ng/L using UPLC-MS for the identification of tetracyclines and sulfonamides. Using magnetic solid-phase extraction with LC-MS/MS, Perez et al. were able to detect antibiotics with detection limits ranging from 11.5 to 26ng/L; however, erythromycin recovery was just 0.84%.

6. METHODS FOR ANTIBIOTIC DISPOSAL:

6.1. For Wastewater:

6.1.1. Anaerobic digestion:

In example, anaerobic digestion (AD) is a practical¹³ and economical way to remove antibiotics from wastewater, especially when treating wastewater from pigs. Antibiotics, however, can lessen the amount of methane produced by AD processes. Such wastewater frequently contains macrolides, tetracyclines, and sulphonamides as antibiotics. To address these pollutants, biosorption and biodegradation are both used. It has been demonstrated that magnetic nanoparticles can eliminate 67% of ciprofloxacin and other contaminants, whereas carbon-based compounds promote bacterial contact and breakdown. Straw is one of the co-substrates that enhances antibiotic elimination through adsorption and co-metabolism. In systems like anaerobic sequencing batch reactors, high removal efficiency for antibiotics like as tetracycline, sulfamethoxazole, azithromycin, and tylosin has been found. These studies demonstrate how AD technology can be used to effectively remove antibiotics from wastewater.

6.1.2. Constructed wetlands:

Constructed wetlands (CWs) effectively remove multi-pollutant antibiotics from swine wastewater, achieving 85-99% removal of oxytetracycline and ciprofloxacin, with vertical flow configurations providing the best results. Antibiotics are immobilized and absorbed by plants during degradation, making CWs more efficient than conventional methods like activated sludge processes. While traditional treatments require more land and energy, CWs offer a more sustainable option. In urban wastewater, CWs removed 61% of doxycycline, 96% of trimethoprim, and 60% of sulfamethoxazole. Despite needing large areas and longer retention times, CWs combined with advanced treatment methods present a promising solution for wastewater remediation.

6.1.3. Algae-based technologies:

The removal of antibiotics from wastewater¹⁴ through processes like adsorption, biodegradation, photodegradation, and hydrolysis may be possible with algae-based technology, according to recent research. A study on the technique of algae-activated sludge revealed that it may eliminate up to 75% of cefradine in a day. When paired with other methods like phytoremediation and anaerobic digestion, this microalgae-based strategy has a lot of promise for getting rid of refractory antibiotics. This might result in a cost-effective and efficient antibiotic removal process.

6.1.4. Application of Biochar:

Wastewater can effectively be treated with biochar, which is made from plant-based materials including rice husk, bamboo, pinewood, and even chicken bones. In one study, hydrogen peroxide (H₂O₂) and biochar made from cotton straw collaborated to remove sulphonamides from a synthetic urine matrix by 60%. The investigation also discovered that while sulphonamides were stable in the absence of biochar, they degraded in the presence of H₂O₂. These results imply that using biochar to remove antibiotics from different environmental matrices can be a sustainable option.

6.1.5. Bioelectrochemical systems:

The removal of antibiotics¹⁵from wastewater through oxidation, reduction, and co-metabolic processes has showed promise for bioelectrochemical systems (BESs), such as microbial fuel cells (MFCs) and microbial electrolysis cells (MECs). An algal-bacterial photo-bio electrochemical system (ABPBS) combined with a solar photovoltaic capacitor, for instance, greatly increased the rates at which florfenicol degraded; increases of 44%, 89%, and 582% were seen at varying capacitor levels. According to a different study, sulfamethoxazole removal rates in closed-circuit upflow anaerobic bio-electrochemical systems (UBES) were 73.7%, which is greater than in open-circuit systems. Current density was a major factor in the enhanced removal of sulfamethoxazole by MFCs operating in plug flow mode. The biodegradation of tetracycline in wastewater was also accelerated by 1.44 times by using bioelectrochemical systems. High quantities of antibiotics can adversely impact biofilm durability and stability despite these encouraging findings.

6.2. Antibiotic isolation from soil:

Due to adsorption and interaction with solid particles, levels of antibiotics in topsoil and bottom deposits are frequently higher than in water. Antibiotic concentrations in soil have been reported to range from 0.5 to 900mg/kg. Since soil micro- and nanopores impede microbial and enzyme diffusion, the bioavailability and bioaccumulation of antibiotics in soil are inversely correlated with their persistence in the soil. There aren't many studies on removing antibiotics from soil in India; instead, most studies concentrate on separating and characterizing antibiotics in soil. For instance, a study conducted in Chennai discovered that wet sewage sludge had higher concentrations of fluoroquinolones, such as norfloxacin, ciprofloxacin, and ofloxacin, than dry sludge. This finding suggests that moisture content raises the levels of antibiotics. An additional study conducted in Assam examined the bacterial populations and soil characteristics, highlighting the potential.

6.3. Other methods for antibiotic degradation:

6.3.1. Bismuth-based photocatalysts for antibiotic degradation:

Bismuth-based photocatalysts play a crucial role in the photocatalytic degradation of antibiotics through a multi-step process. The mechanism begins when the photocatalyst absorbs photons, exciting electrons from the valence band to the conduction band. This separation¹⁷ of photoinduced electrons and holes allows for the degradation of antibiotics. The holes directly attack the antibiotics or generate hydroxyl radicals (•OH) through oxidation of H₂O/OH⁻, which further degrade the antibiotics. Hydroxyl radicals, with a high oxidation potential, are key to breaking down contaminants, especially through adsorption and reaction on the catalyst surface.

The main degradation pathway involves the hydroxyl radicals reacting with the adsorbed antibiotics, leading to their breakdown into smaller molecules like CO₂ and H₂O. Other reductive pathways, such as electron reduction of O₂ into superoxide radicals (O₂⁻), can also occur, but these are less efficient for antibiotic degradation.

The degradation of antibiotics like sulfamethoxazole (SMZ) involves breaking specific chemical bonds, such as the S–N bond, through radical attacks. Other common reactions include the destruction of oxazole rings and hydroxylation of benzene rings. Overall, the photocatalytic process results in the degradation of antibiotics into smaller compounds, aiding in the purification of wastewater.

6.3.2. Semiconductor Assisted Photocatalysis:

Photocatalysis is a process that speeds up chemical reactions using light and a catalyst, without the catalyst being consumed in the process. The term "photocatalysis" is derived from the Greek words for "light" (photo) and "acceleration" (catalysis). It was first demonstrated by Fujishima and Honda in 1972, when they used a TiO₂ electrode under UV light to split water. In the context of wastewater treatment, photocatalysis involves generating free radicals that interact with organic pollutants, causing their breakdown.

The process operates by exposing a semiconductor to light with energy equal to or exceeding its bandgap, which excites electrons from the valence band to the conduction band, leaving behind holes in the valence band. These holes react with water to form hydroxyl radicals, powerful oxidizers that help degrade organic substances. Simultaneously, the excited electrons in the conduction band react with oxygen to create superoxide anions. Together, these oxidation and reduction reactions contribute to the degradation of contaminants in wastewater.

6.3.3. Antibiotic Degradation during Manure Composting:

Reducing the concentration of antibiotics in manure prior to land application through composting is a useful and economical technique that can help mitigate environmental hazards. Manure's odor is mostly controlled and pathogens are managed by this controlled aerobic process, in which microorganisms break down organic compounds at temperatures frequently higher than 40°C. Moreover, contamination from chemicals such as pesticides, hormones, explosives, hydrocarbons, and personal care items can be cleaned up by composting. Studies indicate a considerable decrease in antibiotic levels even though there is little information on how antibiotics degrade during composting. For instance, 58–82% of the sulfachlorpyrazine was reduced in 8 days by composting poultry manure, and this reduction continued for 3 months subsequent to storage. In a similar vein, over 35 days, composting beef manure removed almost 99% of the oxytetracycline, while room temperature only greatly reduced it.

6.3.4. Degradation of antibiotics by Advanced oxidation processes (AOPs):

Antibiotics have emerged as contaminants due to their widespread production and use, posing threats to both ecosystems and human health. Various methods have been explored to remove antibiotics from water and wastewater, including biological, physical, and chemical approaches. Among these, advanced oxidation processes (AOPs) are gaining attention for their fast reaction rates and strong oxidation capabilities, making them effective in degrading antibiotics in aquatic environments. Key AOP techniques include:

a. Fenton and Fenton-like processes: This method involves the combination of ferrous salt and hydrogen peroxide to produce hydroxyl radicals (OH·), which can oxidize organic pollutants in wastewater.

b. Ozonation or catalytic ozonation: Ozone, a powerful oxidizing agent, can directly degrade organic pollutants. It can also react with water in the presence of a catalyst to form hydroxyl radicals, enhancing its oxidation power.

c. Photocatalytic oxidation: Semiconductor materials like TiO₂, ZnS, and SnO₂ act as photocatalysts. When exposed to light, they generate electrons and holes, which lead to the formation of reactive oxygen species like superoxide radicals (O₂·) and hydroxyl radicals (OH·), helping to break down pollutants.

d. Electrochemical oxidation: This method uses an electric current to oxidize organic substances, transforming them into non-toxic products. It includes both direct and indirect oxidation processes.

e. Ionizing radiation: Gamma rays and electron beams can degrade organic pollutants by generating active species that break down contaminants through direct and indirect interactions.

These techniques offer promising solutions for effectively removing antibiotics from water, though the efficiency and suitability of each method can vary based on the specific pollutants and conditions.

7. CONCLUSION:

As antibiotic resistance continues to grow as a global health threat, the environmental impact of widespread antibiotic use in agriculture and aquaculture has gained increasing attention. The improper disposal of antibiotics and their excessive use in animal husbandry are driving environmental contamination, leading to antibiotic-resistant bacteria in natural environments. These resistant bacteria, once established, can spread into human populations through various routes, including contaminated food, drinking water, or direct contact.

There is an urgent need for better regulation and management of antibiotic use in agriculture, as well as more robust research to quantify the full environmental and public health risks. Addressing these challenges requires a multi-faceted approach that involves improving agricultural practices, implementing stricter regulatory measures, and investing in research to develop safer alternatives to antibiotics in farming and aquaculture. This will help mitigate the negative effects on both the environment and public health while ensuring that antibiotics remain effective in treating infections for generations to come.

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Received on 15.10.2024 Revised on 09.12.2024

Accepted on 23.01.2025 Published on 27.02.2025

Available online from March 05, 2025

Asian J. Pharm. Tech. 2025; 15(1):83-89.

DOI: 10.52711/2231-5713.2025.00014

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.