Disease In Chickens Research Articles

Phytochemicals: a promising approach to control infectious bursal disease.

Infectious bursal disease (IBD) is one of the dangerous diseases of poultry that affects the bursa of Fabricius, which is an important organ of the bird's immune system. IBD virus is resistant to many drugs, making its control difficult. Vaccination of IBD is in practice for a long time worldwide to control IBD, but secondary issues like vaccine failure and lower efficacy lead to their reduced use in the field. Multiple medicines are currently used, but the phytochemicals have emerged as promising agents for controlling IBD. The drugs to be developed should possess direct antiviral properties by targeting viral entry mechanisms, enhancing the host immune response, and inhibiting viral protein synthesis. Phytochemicals have potential to contribute to food security by minimizing the possibility of disease outbreaks and ensuring that consumers worldwide obtain healthy poultry products. It has been now claimed that direct and indirect activities of phytochemicals can be effective in the control of IBDV. Although available evidence suggest that the phytochemicals can contribute in controlling occurrence IBDV, there is a definite need of focused studies to gain more insight and develop rational strategies for their practical use. This review highlights the disease caused by IBDV, inhibition of viral replication, boosting the immune system, disruption of viral membrane, and important phytochemicals showing antiviral activities against IBDV.

Development of fatty liver disease model using high cholesterol and low choline diet in white leghorn chickens

Nonalcoholic fatty liver disease (NAFLD), which shows similar symptoms as fatty liver hemorrhage syndrome (FLHS) in chickens, is the most common cause of chronic liver disease and cancer in humans. NAFLD patients and FLHS in chickens have demonstrated severe liver disorders when infected by emerging strains of human hepatitis E virus (HEV) and avian HEV, respectively. We sought to develop a fatty liver disease chicken model by altering the diet of 3-week-old white leghorn chickens. The high cholesterol, and low choline (HCLC) diet included 7.6% fat with additional 2% cholesterol and 800 mg/kg choline in comparison to 5.3% fat, and 1,300 mg/kg choline in the regular diet. Our diet induced fatty liver avian model successfully recapitulates the clinical features seen during NAFLD in humans and FLHS in chickens, including hyperlipidemia and hepatic steatosis, as indicated by significantly higher serum triglycerides, serum cholesterol, liver triglycerides, cholesterol, and fatty acids. By developing this chicken model, we expect to provide a platform to explore the role of lipids in the liver pathology linked with viral infections and contribute to the development of prophylactic interventions.

Occurrence of Salmonellae in Chickens from some Selected Poultry Farms in Kano, Nigeria

Study’s Excerpt/Novelty This study provides a novel investigation into the prevalence of Salmonella species in poultry from selected farms in Kano, Nigeria, using comprehensive random sampling techniques across multiple local government areas. The findings reveal a notably low prevalence of asymptomatic Salmonella infections (1.54%) among chickens, potentially attributable to effective hygiene practices, biosecurity measures, and biological control of vectors. This research contributes valuable baseline data for the epidemiological understanding and management of Salmonella in poultry farms within the region. Full Abstract Infections with bacteria of the genus Salmonella are responsible for a variety of acute and chronic diseases in poultry. This study was carried out to determine the prevalence of Salmonella species in chickens from some selected poultry farms in Kano, Nigeria. Random sampling method was used in the study. One (1) farm was chosen from each of the eight (8) selected Local Government areas of the State. Thirteen (13) samples each from cloacae, egg, dust, drag and boots which is (65) from each farm) were collected from apparently healthy chickens (layers), transported immediately to the laboratory for analyses. The colonies were subsequently subjected to Gram-staining and microscopy. Eight Salmonella isolates were determined, three (3) were isolated from boot swabs, two (2) from cloacal swabs and one (1) each from egg, dust and drag swabs. All the isolates were confirmed by serological test; using Salmonella “O” antigen; group A-S (Oxoid). Asymptomatic Salmonella infection among chickens in the sampled farms is low with (1.54%). The low occurrence could be attributed to proper hygienic practices, biosecurity measure as well as application of biological control of rodents and other vehicles of Salmonella.

Current Status of Poultry Recombinant Virus Vector Vaccine Development.

Inactivated and live attenuated vaccines are the mainstays of preventing viral poultry diseases. However, the development of recombinant DNA technology in recent years has enabled the generation of recombinant virus vector vaccines, which have the advantages of preventing multiple diseases simultaneously and simplifying the vaccination schedule. More importantly, some can induce a protective immune response in the presence of maternal antibodies and offer long-term immune protection. These advantages compensate for the shortcomings of traditional vaccines. This review describes the construction and characterization of primarily poultry vaccine vectors, including fowl poxvirus (FPV), fowl adenovirus (FAdV), Newcastle disease virus (NDV), Marek's disease virus (MDV), and herpesvirus of turkey (HVT). In addition, the pathogens targeted and the immunoprotective effect of different poultry recombinant virus vector vaccines are also presented. Finally, this review discusses the challenges in developing vector vaccines and proposes strategies for improving immune efficacy.

The Status of Indigenous Chicken Genetic Resources: An Analysis of Farmers’ Perspectives and Implications for Breed Conservation Priorities in Zambia

(1) Background: Indigenous chickens (Gallus domesticus) (ICs) are an essential component of agriculture and rural livelihood among 80% of small-scale farmers (SSFs) in Sub-Saharan Africa (SSA). However, in the past decade, significant losses of Indigenous Chicken Animal Genetic Resources (IC-AnGR) resulting from poultry diseases, imported exotic breeds, poor market access, and uncontrolled crossbreeding have been reported. This study aimed to investigate the status of IC-AnGR and assess the major challenges affecting the production of ICs. (2) Methods: We surveyed 358 households in eastern, central, and southern livelihood zones, comprising 81.6% males and 17.6% females. (3) Results: Our study shows that respondents owned 16,112 ICs, 3026 goats, and 5183 herds of cattle. Overall, 77.4% of chicken breeds were ICs and 22.6% were exotic. Across the three zones, 18–44% reported the introduction or adoption of exotic breeds in the past decade, with most households sourcing breeding stock from local communities and family and friends at 45% and 28.6%, respectively. Farmers gave various reasons for adopting new chicken breeds, including fast growth (21.7%), larger mature sizes (21.66%), and resistance to diseases at 15.2%. Overall, 92.5% of farmers agreed or strongly agreed that some IC breeds disappeared in the past decade and nearly 90% were concerned. Some attributed the loss to poultry diseases. (4) Conclusion: Deliberate policies to promote the sustainable use and conservation of ICs are critical in Zambia.

Long-Term Protection against Virulent Newcastle Disease Virus (NDV) in Chickens Immunized with a Single Dose of Recombinant Turkey Herpesvirus Expressing NDV F Protein.

Newcastle disease (ND) is a significant infectious disease in poultry, causing substantial economic losses in developing countries. To control ND, chickens must be vaccinated multiple times a year. In order to develop an improved vaccine that provides long-term protection, the F gene from genotype VII NDV was inserted into the herpesvirus of turkey (HVT) vaccine virus using CRISPR/Cas9-mediated NHEJ repair and Cre/LoxP technology. The immunogenicity and protective efficacy of the resulting recombinant vaccines were evaluated through antibody assays and virus challenge experiments. Two recombinant vaccines, rHVT-005/006-F and rHVT-US2-F, were generated, both exhibiting growth rates comparable with those of HVT in vitro and consistently expressing the F protein. One-day-old specific pathogen-free (SPF) chickens immunized with 2000 PFU/bird of either rHVT-005/006-F or rHVT-US2-F developed robust humoral immunity and were completely protected against challenge with the NDV F48E8 strain at 4 weeks post-vaccination (wpv). Furthermore, a single dose of these vaccines provided sustained protection for at least 52 wpv. Our study identifies rHVT-005/006-F and rHVT-US2-F as promising ND vaccine candidates, offering long-term protection with a single administration. Moreover, HVT-005/006 demonstrates promise for accommodating additional foreign genes, facilitating the construction of multiplex vaccines.

An enhanced image based mobile deep learning model for identification of Newcastle poultry disease

An enhanced mobile deep learning model based on images is presented in this paper to identify Newcastle poultry disease. A dataset of manually annotated and labeled images of the disease was utilized to pre-train an image-based Convolutional Neural Network (CNN). An Android smartphone app was developed to communicate with the model. A local server was integrated with the generated model to do image classification. A mobile application was developed and made available, enabling users to upload a fecal photograph to a website housed on the streamlet server and obtain the model's processed findings. The user regains control over their health status. The model achieved an accuracy of 95% on the test set and was able to correctly identify specific instances of Newcastle poultry disease. The paper discusses the advantages of a mobile-based approach in comparison to traditional methods of identification and proposes the model as an effective low-cost solution for farmers and researchers.

Advancing livestock and poultry disease diagnosis with high-resolution melt curve analysis

High-Resolution Melting (HRM) is a sensitive polymerase chain reaction-based molecular assay used to detect, single nuclear polymorphisms, mutations, and variations in genotypes of pathogens. HRM analysis is more convenient than other detection and discrimination approaches since it is a closed-tube method, that is the polymerase chain reaction amplification and subsequent analysis are carried out sequentially in the well. It finds application in various veterinary diagnoses and discrimination of genotypes of the microorganisms. Numerous public health organizations have recommended increased monitoring activities to support current surveillance programs in response to the threat of bioterrorism and emerging infectious diseases. HRM assay will be an efficient, cost-effective disease diagnostic and genotype of circulating pathogen identification method thereby aiding in disease surveillance which is important for monitoring the spread of infectious diseases. This review emphasizes the application of HRM in livestock, poultry, and companion animals in recent times carried out all over the world for determining the genotype, strain, and breed differentiation including pathogens classification.

The full genome characterization of avian encephalomyelitis virus, Iran: a vertical transmission case.

Avian encephalomyelitis (AE) is an important infectious poultry disease worldwide that is caused by avian encephalomyelitis virus (AEV). The causative virus can be transmitted both horizontally and vertically. In the present study, an AEV suspected outbreak with typical neurological signs occurred in broilers. Histopathological examination, RT-PCR assay and full genome sequencing were applied to confirm the presence of AEV. Phylogenetic analysis of the full genome sequence showed that the detected AEV strain at 7055 nucleotide length is classified in cluster I and is closely related to vaccinal USA and China originated isolates. Although, the outbreaks of AE in progeny of vaccinated breeders have been reported previously, the source of infection was unknown. Based on the results obtained in this study, the outbreaks are vaccine-originated. This study provides the first whole genome analysis of AEV from Iran and reveals that the AEV possesses a hepatitis C virus-like internal ribosome entry site.

Stable isotope profiling and its potential applications to poultry sciences

Metabolomics is a bottom-up approach to studying metabolism in living cells, tissues, organoids, or even whole organisms. In conjunction with genomics and proteomics, it is a crucial tool for better understanding complex biological systems. In comparison to genomics or proteomics, metabolomics more directly reflects the phenotype since it encompasses events that occur downstream of gene expression. In the field of poultry or avian diseases, this method is now well established, although it could have more innovative applications in the future. For example, 13C-guided metabolomics and flux analysis could be used to identify biochemical pathways associated with heat or immune stress. It could also help to explore the effects of alternatives to antimicrobials on the intestinal health of poultry and their action on avian pathogens, including the mechanisms of resistance and persistence. In this review, we discuss some basic principles of metabolomics and its current and potential applications in poultry.

Delineation of chicken immune markers in the era of omics and multicolor flow cytometry

Multiparameter flow cytometry is a routine method in immunological studies incorporated in biomedical, veterinary, agricultural, and wildlife research and routinely used in veterinary clinical laboratories. Its use in the diagnostics of poultry diseases is still limited, but due to the continuous expansion of reagents and cost reductions, this may change in the near future. Although the structure and function of the avian immune system show commonalities with mammals, at the molecular level, there is often low homology across species. The cross-reactivity of mammalian immunological reagents is therefore low, but nevertheless, the list of reagents to study chicken immune cells is increasing. Recent improvement in multicolor antibody panels for chicken cells has resulted in more detailed analysis by flow cytometry and has allowed the discovery of novel leukocyte cell subpopulations. In this article, we present an overview of the reagents and guidance needed to perform multicolor flow cytometry using chicken samples and common pitfalls to avoid.

Supplementation with organic yeast-derived selenium provides immune protection against experimental necrotic enteritis in broiler chickens

Necrotic enteritis (NE) is a potentially fatal poultry disease that causes enormous economic losses in the poultry industry worldwide. The study aimed to evaluate the effects of dietary organic yeast-derived selenium (Se) on immune protection against experimental necrotic enteritis (NE) in commercial broilers. Chickens were fed basal diets supplemented with different Se levels (0.25, 0.50, and 1.00 Se mg/kg). To induce NE, Clostridium perfringens (C. perfringens) was orally administered at 14 days of age post hatch. The results showed that birds fed 0.25 Se mg/kg exhibited significantly increased body weight gain compared with the non-supplemented/infected birds. There were no significant differences in gut lesions between the Se-supplemented groups and the non-supplemented group. The antibody levels against α-toxin and NetB toxin increased with the increase between 0.25 Se mg/kg and 0.50 Se mg/kg. In the jejunal scrapings and spleen, the Se-supplementation groups up-regulated the transcripts for pro-inflammatory cytokines IL-1β, IL-6, IL-8, iNOS, and LITAF and avian β-defensin 6, 8, and 13 (AvBD6, 8 and 13). In conclusion, supplementation with organic yeast-derived Se alleviates the negative consequences and provides beneficial protection against experimental NE.

OASL suppresses infectious bursal disease virus replication by targeting VP2 for degrading through the autophagy pathway.

OASL (2´,5´-oligadenylate synthetase-like protein) exhibits broad-spectrum antiviral effects against single-stranded RNA viruses in mammals, potentially serving as a promising target for novel antiviral strategies. However, its role in inhibiting the replication of double-stranded RNA viruses (dsRNA viruses), such as infectious bursal disease virus (IBDV), in avian species remains unclear. Our findings indicated a unique and potent defense mechanism of OASL against dsRNA viruses. It has been previously shown in mammals that OASL inhibits virus replication through increasing interferon production. The groundbreaking aspect of our study is the finding that OASL has the ability to interact with IBDV viral protein VP2 and target it for degradation and thus exerts its antiviral effect. Our results reveal the interaction between avian natural antiviral immune response and IBDV infection. Our study not only enhances our understanding of bird defenses against viral infections but can also inform strategies for poultry disease management.

Simultaneous construction strategy using two types of fluorescent markers for HVT vector vaccine against infectious bursal disease and H9N2 avian influenza virus by NHEJ-CRISPR/Cas9.

Recently, herpesvirus of turkeys (HVT), which was initially employed as a vaccine against Marek's disease (MD), has been shown to be a highly effective viral vector for producing recombinant vaccines that can simultaneously express the protective antigens of multiple poultry diseases. Prior to the development of commercial HVT-vectored dual-insert vaccines, the majority of HVT-vectored vaccines in use only contained a single foreign gene and were often generated using time-consuming and inefficient traditional recombination methods. The development of multivalent HVT-vectored vaccines that induce simultaneous protection against several avian diseases is of great value. In particular, efficacy interference between individual recombinant HVT vaccines can be avoided. Herein, we demonstrated the use of CRISPR/Cas9 gene editing technology for the insertion of an IBDV (G2d) VP2 expression cassette into the UL45/46 region of the recombinant rHVT-HA viral genome to generate the dual insert rHVT-VP2-HA recombinant vaccine. The efficacy of this recombinant virus was also evaluated in specific pathogen-free (SPF) chickens. PCR and sequencing results showed that the recombinant virus rHVT-VP2-HA was successfully constructed. Vaccination with rHVT-VP2-HA produced high levels of specific antibodies against IBDV (G2d) and H9N2/Y280. rHVT-VP2-HA can provide 100% protection against challenges with IBDV (G2d) and H9N2/Y280. These results demonstrate that rHVT-VP2-HA is a safe and highly efficacious vaccine for the simultaneous control of IBDV (G2d) and H9N2/Y280.

Molecular Detection and Characterization of Newcastle Disease Virus from Chickens in Mid-Rift Valley and Central Part of Ethiopia.

Newcastle disease (ND) is a highly infectious poultry disease that causes major economic losses worldwide. The disease is caused by Newcastle Disease Virus (NDV) and early detection and identification of the viral strain is essential. Having knowledge of the NDV strain genotype that circulates in some regions would help in designing an effective vaccine to control the disease. In this regard, there is little information on NDV strain in chickens in mid Rift Valley and the central part of Ethiopia. Therefore, the purpose of this study was to detect and characterize NDV strain genotype from chickens in mid-Rift Valley and the central part of Ethiopia and test whether this NDV strain genotype matches the vaccine strain currently used in the study area. A total of 98 samples: 78 (tracheal and cloacal) swabs from chicken pools of five and 20 tissue samples were collected. To detect NDV strain, conserved region of the virus Matrix (M) gene was amplified by qRT-PCR. To characterize NDV strain genotypes, M-gene positive samples were specifically re-amplified by conventional PCR targeting the Fusion (F) gene region and sequenced by Sanger method. 13.26% of tested samples were positive for NDV strain in the study area with statistically significant difference (P<0.05) among the study sites. Further characterization of the F genes from NDV strain isolates by phylogenetic analysis indicated that one field isolate clustered with genotype VII whereas three of the isolates clustered to genotype I, II, and III. The isolate of the current NDV strain vaccine in use in the study area clustered with genotype II. The current study indicates the existence of different NDV strain genotype from that of the vaccine strain currently used. Even though large-scale characterization of several isolates is required at national level, the current study laid baseline information for the existence of variations between field NDV strain genotype and vaccine strain currently used against ND in the country.

Ethnoveterinary practice of poultry birds in Ilara-mokin, Ondo-state Nigeria

Ethnoveterinary practices are important because they are easily available, inexpensive and effective, especially in rural areas where veterinary services are absent or irregular and expensive. The need to obtain information on ethnoveterinary practice in poultry in Ilara-mokin in Ondo State Nigeria formed the basis of the study. The study investigated the ethnoveterinary practice in poultry in Ilara-mokin, Ondo State Nigeria. Data used were collected from rural poultry farmers in the community using an open ended interview with the use of questionnaire and were analysed by means of simple average and percentages. The study revealed that female dominated 83.3% rural poultry production within age of 36-50 (50%) and no formal education (50%) and were mostly Christians (100%) and traders (55.56%). The ethnoveterinary practice in the study area include medicinal plants, palm oil, fire and so on. Seven medicinal plants belonging to seven families were discovered. The leaves, roots and fruit from the medicinal plants were used for treating poultry diseases. Ten diseases were discovered in poultry birds. Indigenous knowledge of the diseases in poultry birds were recorded. The economy of the state can be improved by increasing the health status of poultry birds.

Artemisia annua L. polysaccharide improves the growth performance and intestinal barrier function of broilers challenged with Escherichia coli.

With the high intensification of poultry breeding, a series of diseases caused by pathogenic bacteria threaten the health of poultry and human. Among them, poultry diseases induced by Escherichia coli cause significant economic loss every year. The aim of this study was to investigate the effects of dietary supplementation with Artemisia annua L. polysaccharide (AAP) on the growth performance and intestinal barrier function of broilers with Escherichia coli (E. coli) challenge. A total of 256 one-day-old chicks were randomly assigned to four treatment groups: control group (fed basal diet), AAP group (fed basal diet supplemented with AAP), E. coli group (fed basal diet and orally administered E. coli), AAP + E. coli group (fed basal diet supplemented with AAP and orally administered E. coli). Dietary AAP supplementation elevated the BW, ADG and ADFI in non-challenged broilers. AAP also increased the apparent metabolic rate of EE and Ca in E. coli-challenged broilers. Moreover, AAP not only enhanced the serum IgA content but also decreased the serum and jejunum content of IL-6, as well as the jejunum level of IL-1β in non-challenged broilers. AAP also down-regulates the mRNA level of inflammatory factors (IL-1β, IL-6, and TNF-α) by inhibiting the mRNA expression of TLR4 and MyD88 in intestinal NF-κB signaling pathway of E. coli-challenged broilers. Meanwhile, AAP up-regulates the activity and mRNA level CAT by down-regulating the mRNA level of Keap1 in intestinal Nrf2 signaling pathway of E. coli-challenged broilers, and decreased serum MDA concentration. AAP significantly elevated the mRNA level of CAT, SOD and Nrf2 in jejunal of non-challenged broilers. Interestingly, AAP can improve intestinal physical barrier by down-regulating serum ET content, increasing the jejunal villus height/crypt depth (VH/CD) and ZO-1 mRNA level in broilers challenged by E. coli. AAP also elevated the VH/CD and the mRNA level of Occludin, ZO-1, Mucin-2 in non-challenged broilers. Importantly, AAP reshaped the balance of jejunum microbiota in E. coli-challenged broilers by altering α diversity and community composition. In summary, AAP ameliorated the loss of growth performance in broilers challenged with E. coli, probably by regulating the intestinal permeability and mucosa morphology, immune function, antioxidant ability, and microbiota.

Isolation of avian influenza virus from backyard poultry population of tehsil Abbottabad, Khyber Pakhtunkhwa, Pakistan

The avian influenza virus is a highly contagious viral disease that predominantly impacts avian species, especially domestic birds like chickens, ducks, and turkeys. The study was conducted to isolate the avian influenza virus from domestic poultry breeds (Desi, Aseel, Fayoumi, and Golden) from backyard poultry population of the Tehsil Abbottabad from January to June 2023. A cross-sectional study was conducted in three rural and five urban areas of Tehsil Abbottabad Using sterile cotton swabs, 128 (tracheal and cloacal) swab samples were collected. The samples were labelled and stored in a brain-heart infusion medium, then transported to the National Reference Laboratory for Poultry Diseases in Islamabad using a thermostat shipment box. The samples were processed using a real-time reverse transcription polymerase chain reaction for avian influenza virus detection. This study concludes with a significant 9.38% prevalence percentage of avian influenza virus in Tehsil Abbottabad backyard poultry. The prevalence percentage of H9 was 83.33%, and that of H5 was 16.66%. The findings reveal the importance of implementing preventative measures to curb avian influenza outbreaks within the poultry population of the district. This study offers vital data for AIV research in 2023, guiding enhanced containment and prevention in backyard poultry to control avian influenza.

Outbreak Of Fowlpox In Commercial Brown Pullets Previously Vaccinated With Fowlpox Vaccine In Zaria, Kaduna State, Nigeria

Fowlpox is a contagious viral disease of poultry, ranked as the third most important cause of mortality in Nigerian indigenous chickens. The present study confirms an outbreak of fowlpox in a flock of 500 Isa brown, 11-weeks-old pullets in Zaria, Kaduna State, Nigeria. The pullets were managed under deep litter system and previously vaccinated with live attenuated vaccine (ABIC biological laboratories Ltd. Israel) at 8-weeks old. They had gross lesions suggestive of cutaneous fowlpox on skin of the head region with 58% morbidity but persistent low mortality rate of 13.8%. Fowlpox virus was isolated from the cutaneous lesions using chorioallantoic membrane (CAM) inoculation in 9 to 11-day-old embryonated chicken eggs based on the presence of opaque-white pock lesions, then confirmed by Agar Gel Immuno Diffusion test using homologous known fowlpox virus (antigen) and antisera. Specific antibodies to fowlpox virus were quantified from the sera of the chickens using an indirect ELISA (Abbkine, Inc, China). The case was managed using; Intramuscular injection of oxytetracycline (TLA), topical applications of oxytetracycline spray as well as oral administrations of Doxygen® and vitamins. Apparently healthy chickens were revaccinated against fowlpox and the pen was thoroughly cleaned, washed, and disinfected. This report had shown an occurrence of vaccine failure, alongside a promising management option and highlighted the significance of vaccination with strict biosecurity measures in the control of fowlpox in Nigeria.

Determination of sulfonamide antibiotics by magnetic porous carbon solid-phase extraction coupled with capillary electrophoresis

Sulfonamide antibiotics (SAs) have been widely used as antibacterial drugs for the prevention and treatment of livestock and poultry diseases, but they seriously threaten human health because they can accumulate in humans. Therefore, it is highly important to develop methods for monitoring sulfonamide residues in aquaculture and food. In this research, based on the generation of porous carbon (PC) by the pyrolysis of sodium citrate, magnetic porous carbon (PC@Fe3O4) was synthesized by a solvothermal method and used as an adsorbent for the magnetic solid-phase extraction of SAs. The effects of the proportion of PC in PC@Fe3O4, adsorbent dosage, adsorption time, eluent type, extraction pH, salt concentration and eluent dosage on the extraction efficiency were systematically studied. The adsorption performance and behavior of PC@Fe3O4 on SAs were evaluated using adsorption kinetics and adsorption isotherms, and the adsorption mechanism was preliminarily discussed. Under optimal conditions, combined with capillary electrophoresis diode array detection, a sensitive detection method for SAs was developed. The proposed method can be used for the determination of six SAs in fishpond water and milk samples, with a linear range of 0.5–200 ng mL−1, detection limits of 0.24–0.34 ng mL−1, and spiked recoveries of 85.9–109.0 %.