Comparison of four different multiclass, multiresidue sample preparation methods in the analysis of veterinary drugs in fish and other food matrices.

In 2018, AOAC International issued Standard Method Performance Requirements (SPMR) 2018.010 - Screening and Identification Method for Regulated Veterinary Drug Residues in Food. In response, we compared 4 different multiresidue methods of sample preparation using the same analytical method entailing ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Tilapia was chosen for testing, and the analytes and monitoring levels were from SPMR 2018.010. The methods consist of efficient procedures with published validation results from the US Department of Agriculture (USDA), Food and Drug Administration (FDA), and Canadian Food Inspection Agency (CFIA), and an enhanced-matrix removal (EMR)-Lipid protocol from China. Each method was used to prepare 102 final extracts of tilapia spiked or not at different levels with the 78 targeted analytes plus metabolites. The same FDA/USDA rules of mass spectral identification were employed in all analyses to assess rates of false positives and negatives. Quantitative accuracy of the methods was also compared in terms of recoveries and reproducibility of spiked tilapia, incurred catfish, and spiked and certified reference material of bovine muscle. Each method yielded generally acceptable results for the targeted veterinary drugs, but the USDA "extract & inject" method was the fastest, simplest, and cheapest to achieve equally or more acceptable results for the widest scope of analytes for the tested food matrices.

Validated spectrophotometric determination of maduramicin ammonium using three charge transfer complexation reactions.

Direct spectrophotometric determination of Maduramicin ammonium (MAD) represents an analytical challenge since it is a weak UV-absorbing and lacking a strong chromophore. This work represents the first spectrophotometric determination of MAD as no direct spectrophotometric or colorimetric determination methods for MAD are available in the literature. The present study illustrates the development of three simple, rapid and inexpensive colorimetric methods for the routine quality control analysis of MAD based on the formation of colored charge transfer complexes with three electron acceptors namely p-chloranilic acid (p-CA), 2,3-dichloro-5,6-dicyano-p-benzoquinone (DDQ) and picric acid (PA). The color products of MAD with p-CA, DDQ and PA were measured at 519, 588 and 405nm respectively. The proposed methods were validated in terms of linearity, ranges, precision, accuracy, robustness and limits of detection and quantification. MAD was effectively determined over concentration ranges of 100-1000, 25-250 and 30-150µg/mL using p-CA, DDQ and PA, respectively with good linearity as shown by the values of correlation coefficients not less than 0.9991. The developed methods were successfully implemented in the assay of MAD powder pharmaceutical formulation for veterinary use.

Nonmedical Uses of Antibiotics: Time to Restrict Their Use?

The global crisis of antibiotic resistance has reached a point where, if action is not taken, human medicine will enter a postantibiotic world and simple injuries could once again be life threatening. New antibiotics are needed urgently, but better use of existing agents is just as important. More appropriate use of antibiotics in medicine is vital, but the extensive use of antibiotics outside medical settings is often overlooked. Antibiotics are commonly used in animal husbandry, bee-keeping, fish farming and other forms of aquaculture, ethanol production, horticulture, antifouling paints, food preservation, and domestically. This provides multiple opportunities for the selection and spread of antibiotic-resistant bacteria. Given the current crisis, it is vital that the nonmedical use of antibiotics is critically examined and that any nonessential use halted.

Quality control programmes for veterinary antimicrobial medicines.

The health benefits of the antimicrobial's use is inherently associated to the risk of antimicrobial resistance (AMR), an ever-increasing multifactorial problem, closely related with injudicious use of antimicrobials, and the lack of new antimicrobial medicines on the market, particularly for veterinary use. Currently, an increasing number of regulatory "One Health" action plans on AMR are running worldwide, already based on monitoring and surveillance systems for resistance and antimicrobials consumption. Such plans are still not mandatory in the European Union member States (EU-MS), but post marketing annual programmes for quality controls of medicines are, to verify and ensure full compliance with the marketing authorizations. The European "risk level" sampling is not based on the conventional risk-ranking process of severity factors vs the probability of occurrence, but instead, on the conviction that in the European Union (EU) all medicines are produced under good manufacturer practices (GMP) and rigorously controlled for quality by the marketing authorization holders (MAH). The present paper links poor-quality antimicrobials and AMR, highlighting examples of regulatory initiatives on this subject outside the EU, particularly those resulting from the World Health Organization (WHO) recommendations. It also intends to trigger a discussion on the role of such quality control programmes, particularly for antimicrobials, beyond the control at any stage of the quality parameters of a marketed medicine, to reflect whether or not it might be relevant to other regulatory coordinated actions against AMR.

National post-market surveillance assessment of veterinary medicines in Korea during the past decade.

BACKGROUND: Veterinary medicines have been widely used for the prevention and treatment of diseases, growth promotion, and to promote feeding efficacy in livestock. As the veterinary medicine industry has steadily grown, it is crucial to set up a baseline for the quality of medicine as well as the insufficiency or excessiveness of the active ingredients in drug products to ensure the compliance, safety and efficacy of these medicines. Thus, the 10 years data of post-marketing quality control study was summarized to determine the rate and extent of non-compliance of these medicines and to establish baseline data for future quality control measures of veterinary medicine. RESULTS: In this study, 1650 drugs for veterinary use were collected per year from each city and province in Korea and analysed for the quantity of active ingredients according to the "national post-market surveillance (NPMS) system" over the past decade. The NPMS assessment was performed using liquid and gas chromatography, titration, UV/Vis spectrophotometry, and bioassays. A total of 358 cases were deemed noncompliant, with the average noncompliance rate for all medicine types being 2.0%. The average noncompliance rates for antibiotics, biologics and other chemical drugs except antibiotics (OCD) were 1.1%, 1.2%, and 3.0%, respectively. The first leading cause for noncompliant products was insufficient quantity of major ingredients (283 cases), and the second leading cause was the existence of excess amount of active ingredients (60 cases). Tylosin, spiramycin, ampicillin, tetracyclines and penicillins were most frequently found to be noncompliant among antibiotics. Among the OCD, the noncompliance was found commonly in vitamin A. CONCLUSION: The overall trend presented gradually decreasing violation rates, suggesting that the quality of veterinary medicines has improved. Consistent application of the NPMS assessment and the establishment of the Korea Veterinary Good Manufacturing Practice (KVGMP) will help to maintain the good quality of medicine.

How to transport veterinary drugs in insulated boxes to avoid thermal damage by heating or freezing.

BACKGROUND: The transport of veterinary drugs must comply with the general standards for drug storage. Although many vehicles are equipped with active heating and/or cooling devices assuring recommended storage conditions, simple insulated transport boxes are also often used. In this study, measurements for typical transport boxes were performed under laboratory conditions by the use of a climate chamber for a temperature of -20 °C and 45 °C to investigate the impact of box size, insulation material, liquid vs. dry filling products, filling degree and other parameters on the thermal performance of insulated boxes. Model calculations and instructions are presented to predict the retention time of recommended drug storage temperatures. RESULTS: The measurements and the model calculations showed that the loading of the transport boxes with additional water bottles to increase the heat capacity is appropriate to prolong the retention time of the recommended temperature range of the drugs. Insulated transport boxes are not suitable to store drugs over a period of more than approximately 12 h. For practical use a recipe is presented to measure the thermal properties of a transport box and the related retention time for which the recommended storage temperatures can be assured. CONCLUSIONS: The following principles for drug transportation in vehicles are recommended: (1) Before transfer into boxes, drugs should always be thermally preconditioned (2) Increase the filling degree of the boxes with thermally preconditioned water bottles or re-usable thermal packs will increase the heat capacity. Do not deep-freeze the bottles or packs below 0 °C to avoid drug freezing due to contact. (3) Open the lid of the boxes only to uncase drugs that are immediately needed. (4) The bigger the box and the higher the filling degree, the longer the retention time of the transport box. (5) Wherever possible, place the drug box at a cool site inside the vehicle. (6) The monitoring of the inside temperature of the transport boxes is recommended. By the proper use of such transport boxes the recommended temperatures can be maintained over one working day.

Recommendations for Clinical Pathology Data Generation, Interpretation, and Reporting in Target Animal Safety Studies for Veterinary Drug Development.

Clinical pathology testing is routinely performed in target animal safety studies in order to identify potential toxicity associated with administration of an investigational veterinary pharmaceutical product. Regulatory and other testing guidelines that address such studies provide recommendations for clinical pathology testing but occasionally contain outdated analytes and do not take into account interspecies physiologic differences that affect the practical selection of appropriate clinical pathology tests. Additionally, strong emphasis is often placed on statistical analysis and use of reference intervals for interpretation of test article-related clinical pathology changes, with limited attention given to the critical scientific review of clinically, toxicologically, or biologically relevant changes. The purpose of this communication from the Regulatory Affairs Committee of the American Society for Veterinary Clinical Pathology is to provide current recommendations for clinical pathology testing and data interpretation in target animal safety studies and thereby enhance the value of clinical pathology testing in these studies.

Requirements for Foreign and Domestic Establishment Registration and Listing for Human Drugs, Including Drugs That Are Regulated Under a Biologics License Application, and Animal Drugs. Final rule.

The Food and Drug Administration (FDA) is amending its regulations governing drug establishment registration and drug listing. These amendments reorganize, modify, and clarify current regulations concerning who must register establishments and list human drugs, human drugs that are also biological products, and animal drugs. The final rule requires electronic submission, unless waived in certain circumstances, of registration and listing information. This rulemaking pertains to finished drug products and to active pharmaceutical ingredients (APIs) alone or together with one or more other ingredients. The final rule describes how and when owners or operators of establishments at which drugs are manufactured or processed must register their establishments with FDA and list the drugs they manufacture or process. In addition, the rule makes certain changes to the National Drug Code (NDC) system. We are taking this action to improve management of drug establishment registration and drug listing requirements and make these processes more efficient and effective for industry and for us. This action also supports implementation of the electronic prescribing provisions of the Medicare Prescription Drug, Improvement, and Modernization Act of 2003 (MMA) and the availability of current drug labeling information through DailyMed, a computerized repository of drug information maintained by the National Library of Medicine.

Antimicrobial resistance and extended-spectrum ß-lactamases of Salmonella enterica serotypes isolated from livestock and processed food in Portugal: an update.

As Salmonella is a common foodborne pathogen, the present study aimed to determine the distribution of Salmonella enterica serotypes isolated during 2011-2012 from poultry, swine, cattle, and processed food in Portugal, and to characterize the antimicrobial susceptibility and the extended-spectrum ß-lactamases (ESBLs). Results were also compared with data obtained before the implementation of the National Control Program in Poultry and the ban of antimicrobial agents in animal feed in the European Union (EU). A total of 14 serotypes were identified, from 258 isolates recovered, with Salmonella Typhimurium (32.6%, n=84) and Salmonella Enteritidis (10.1%, n=26) being the most common. Salmonella Enteritidis in poultry was less frequent than in previous studies, which might be associated with the implementation of the National Control Program for Salmonella in poultry. Nevertheless, other serotypes seem to occupy this biological niche, and may be more common in human salmonellosis in the future. The majority of isolates (70.2%, n=181) were resistant to at least one class of antimicrobial agent and exhibited higher frequency of resistance to tetracycline (47.7%, n=123) and ampicillin (36.0%, n=93), with Salmonella Typhimurium being the more resistant serotype. Resistance to fluoroquinolones was shown in 8% (n=21) of isolates, a lower value compared to data obtained before 2004. ESBLs producers Salmonella Typhimurium bla(CTX-M-1) and Salmonella Enteritidis bla(SHV-12) were isolated from swine and poultry, respectively. The bla(CTX-M-1) and bla(SHV-12) genes were carried on conjugative plasmids of IncHI2replicon types and IncI1, respectively. This was the first report of a bla(CTX-M-1) in Salmonella Typhimurium in Portugal. Overall, the results revealed changes in animal origin Salmonella serotypes, mainly emerging serotypes, in frequency of resistance, and in occurrence of ESBLs-producing Salmonella. The control measures taken by the EU seem to have some impact on the resistance rate of some antibiotics such as quinolones. The emergence of ESBLs and its potential spread among animal reservoirs and the food chain highlight the continuous antimicrobial surveillance at the animal level.

Animal trypanosomosis: making quality control of trypanocidal drugs possible.

African animal trypanosomosis is arguably the most important animal disease impairing livestock agricultural development in sub-Saharan Africa. In addition to vector control, the use oftrypanocidal drugs is important in controlling the impact of the disease on animal health and production in most sub-Saharan countries. However, there are no internationally agreed standards (pharmacopoeia-type monographs or documented product specifications) for the quality control of these compounds. This means that it is impossible to establish independent quality control and quality assurance standards for these agents. An international alliance between the Food and Agriculture Organization of the United Nations, the International Federation for Animal Health, the Global Alliance for Livestock Veterinary Medicines, the University of Strathclyde and the International Atomic Energy Agency (with critical support from the World Organisation for Animal Health) was established to develop quality control and quality assurance standards for trypanocidal drugs, with the aim of transferring these methodologies to two control laboratories in sub-Saharan Africa that will serve as reference institutions for their respective regions. The work of the international alliance will allow development of control measures against sub-standard or counterfeit trypanocidal drugs for treatment of trypanosome infection. Monographs on diminazene aceturate (synonym: diminazene diaceturate), isometamidium chloride hydrochloride, homidium chloride and bromide salts and their relevant veterinary formulations for these agents are given in the annex to this paper. However, the authors do not recommend use of homidium bromide and chloride, because of their proven mutagenic properties in some animal test models and their suspected carcinogenic properties.

[Official testing of immunological veterinary medicinal products]. / Unabhängige Prüfung von immunologischen Tierarzneimitteln.

The testing of immunological veterinary medicinal products (IVMPs) by official medicines control laboratories (OMCLs) is an important contribution to the control of quality, safety and efficacy of these products. Based on the legislation of the European Union (EU) and with the support of the European Directorate for the Quality of Medicines & HealthCare (EDQM) a network of OMCLs of the EU member states and Switzerland has been built. This network has established its own rules allowing the mutual recognition of test results and rapid communication of details regarding batch release or rejection. Annual reports, OMCL meetings and collaborative studies help to foster confidence between the OMCLs. The procedure for official testing is described and an overview of deficits found at testing is presented in the paper. The testing of selected batches of centrally authorized products is also performed by OMCLs and is briefly described. Communication both among OMCLs and with pharmaceutical industry is an important part of the OMCLs' work to compare test results and to optimize existing or develop new test methods. Several OMCLs are also pursuing the development of new test methods, primarily for the reduction, refinement and replacement of animal experiments in routine testing.

Exotic Animal Practice in Africa.

This article explores the evolution, unique aspects, and challenges facing exotic animal practice in South Africa. This article delves into the slow emergence of dedicated exotic practices and the challenges faced by veterinarians in a vast and diverse landscape. The unique nature of the veterinary landscape is highlighted, emphasizing the impact of varied climates on species inhabiting different regions. The challenges are multifaceted, ranging from limited education infrastructure to dietary issues, unregulated feeds, and the complexities of herbal medicine use. The narrative explores client education challenges due to the scarcity of dedicated practices, underlining the importance of communication channels.

An in vitro method for evaluating endotoxic activity using prostaglandin E(2) induction in bovine peripheral blood.

Severe side effects of veterinary vaccines, in particular Histophilus somni-containing vaccines for cows, have frequently been reported in Japan. These side effects are probably caused by endotoxins. Contamination levels of endotoxins could be monitored using the Limulus amebocyte lysate (LAL) test; however, the LAL test is not completely adequate for evaluation of in vivo endotoxic activities. In this study, we established a method for evaluating endotoxic activities using prostaglandin E2 (PGE2) induction in bovine peripheral blood. Blood and standard endotoxin, derived from Escherichia coli, were mixed and incubated. The concentration of induced PGE2 in the culture supernatant reached a maximum after 24-h incubation. A linear dose-response curve was observed for PGE2 concentration and the logarithmic transformed standard endotoxin concentration (5-5000 ng/ml). The endotoxic activity of H. somni in cows was the highest among those of several tested endotoxins. However, the LAL activities of H. somni were not as high as those of the other tested endotoxins. These results may provide a reason for the many report of side effects of H. somni-containing vaccines. The PGE2 detection assay described here could be a valuable method for evaluating the endotoxic activities of vaccines in cows.

History of chronic toxicity and animal carcinogenicity studies for pharmaceuticals.

During the 20th century, as drug products were being developed to treat both known and emerging human diseases and conditions, determining the safety of these new chemicals became of increasing importance and necessity. For a time, the safety of use in human populations was of question, let alone whether the drug product was truly effective. As such, US and international regulatory agencies have played a major role in establishing standardized testing to evaluate the safety and efficacy of drug products. Pharmacologic and toxicologic evaluation of a new drug in animals is an important part of the pharmaceutical development process prior to its first-time use in humans, as well as its potential chronic use in affected populations. Just as both science and technology have evolved over the past century and further, so have the guidelines that have been put forth to adequately and efficiently evaluate the toxicity of new drugs and their subsequent safety in humans. This review summarizes the historical highlights of the conduct of drug safety evaluations in animals, particularly with regard to chronic toxicity and carcinogenicity assessments, and how we have progressed to our current standards and protocols to ensure safe use of drug products in human populations.

A refined approach to estimate exposure for use in calculating the Maximum Residue Limit of veterinary drugs.

Maximum Residue Limits (MRLs) are standards that represent the maximum residue concentration expected to be found if a veterinary drug is administered according to good practice in the use of veterinary drugs (GVP). MRLs are established only where the exposure to residues in food resulting from particular use patterns of the veterinary drug pass a public health risk assessment. The current model diet as used by major regulators overstates mean consumption of food for populations when compared to results from food surveys of actual consumption. Exposure to residues is overestimated when calculating long-term (chronic) exposure using the model diet leading to the risk to consumers being overstated. Additionally the model diet underestimates the size of large portions eaten by the group of consumers that eat large quantities of a particular food in a single meal potentially leading to understating of risks associated with exposure to residues of drugs that produce an adverse effect after a single exposure. A revision of dietary consumption figures is proposed that will better match the consumption figures used in point-estimates of dietary exposure to the timeframe for consumption that is relevant to the reference dose.

Recent progress and future directions for reduction, refinement, and replacement of animal use in veterinary vaccine potency and safety testing: a report from the 2010 NICEATM-ICCVAM International Vaccine Workshop.

Veterinary vaccines contribute to improved animal and human health and welfare by preventing infectious diseases. However, testing necessary to ensure vaccine effectiveness and safety can involve large numbers of animals and significant pain and distress. NICEATM and ICCVAM recently convened an international workshop to review the state of the science of human and veterinary vaccine potency and safety testing, and to identify priority activities to advance new and improved methods that can further reduce, refine and replace animal use. Rabies, Clostridium sp., and Leptospira sp. vaccines were identified as the highest priorities, while tests requiring live viruses and bacteria hazardous to laboratory workers, livestock, pets, and wildlife were also considered high priorities. Priority research, development and validation activities to address critical knowledge and data gaps were identified, including opportunities to apply new science and technology. Enhanced international harmonization and cooperation and closer collaborations between human and veterinary researchers were recommended to expedite progress. Implementation of the workshop recommendations is expected to advance new methods for vaccine testing that will benefit animal welfare and ensure continued and improved protection of human and animal health.

Potency testing of veterinary rabies vaccines: replacement of challenge by in vitro testing: considerations for development of alternative assays.

Vaccination of domestic animals against rabies creates a critical barrier between wildlife reservoirs and the human population. Ensuring these vaccines are potent and effective is paramount in preventing human exposure to this deadly and costly disease. The National Institutes of Health (NIH) test is, at present, the most widely used and internationally recommended potency assay for batch testing inactivated rabies vaccines. This test has numerous inherent limitations and disadvantages, including a lack of precision. The NIH test requires a large number of animals and involves unrelieved pain and suffering. A relevant in vitro assay should provide a more accurate, reproducible, rapid, safe, and humane rabies vaccine potency test.

The validation of potency tests: hurdles identified by EMA/CVMP/IWP.

The biological nature of IVMPs leads to some unavoidable batch to batch variation in production. The potency test is part of the quality control of the finished product intended to confirm consistency of production and that each batch is formulated equivalent to batches that have been demonstrated to be efficacious. Adequate validation of potency tests is essential to ensure that the results of the assays accurately reflect the amount, titre, or potency of the active substance measured and to indicate the limitations on the accuracy of the measurements to be expected from the test used. The CVMP/IWP published their conclusions concerning validation of potency tests in a Reflection Paper in March 2010. The test validation must demonstrate a dose response and the precision of the result should enable reliable detection of a sub-standard batch. However, the inherent variability in experimental animals often leads to unacceptably wide confidence intervals for in vivo tests which limits their ability to detect slight changes of the antigen amount. The development of in vitro methods as alternatives to in vivo potency tests is encouraged.

In vitro vaccine potency testing: a proposal for reducing animal use for requalification testing.

This paper proposes a program under which the use of animals for requalification of in vitro potency tests could be eliminated. Standard References (USDA/CVB nomenclature) would be developed, characterized, stored and monitored by selected reference laboratories worldwide. These laboratories would employ scientists skilled in protein and glycoprotein chemistry and equipped with state-of-the-art instruments for required analyses. After Standard References are established, the reference laboratories would provide them to the animal health industry as "gold standards". Companies would then establish and validate a correlation between the Standard Reference and the company Master Reference (USDA/CVB nomenclature) using an internal in vitro assay. After this correlation is established, the company could use the Standard References for qualifying, monitoring and requalifying company Master References without the use of animals. Such a program would eliminate the need for animals for requalification of Master References and the need for each company to develop and validate a battery of Master Reference Monitoring assays. It would also provide advantages in terms of reduced costs and reduced time for requalification testing. As such it would provide a strong incentive for companies to develop and use in vitro assays for potency testing.