Vaccine Production Facilities Research Articles

Determination of Optimal Antigen Yield and Virus Inactivation Conditions for the Production of the Candidate Foot-and-Mouth Disease Recombinant Vaccine Strain Asia1 Shamir-R in a Bioreactor.

Since the foot-and-mouth disease (FMD) outbreak in South Korea in 2010-2011, vaccination policies utilizing inactivated FMD vaccines composed of types O and A have been implemented nationwide. However, because type Asia1 occurred in North Korea in 2007 and intermittently in neighboring countries, the risk of type Asia1 introduction cannot be ruled out. This study evaluated the antigen yield and viral inactivation kinetics of the recombinant Asia1 Shamir vaccine strain (Asia1 Shamir-R). When Asia1 Shamir-R was proliferated in shaking flasks (1 L), a 2 L bioreactor (1 L), and a wave bioreactor (25 L), the antigen yields were 7.5 μg/mL, 5.2 μg/mL, and 3.8 μg/mL, respectively. The optimal FMDV inactivation conditions were 2 mM BEI at 26 °C and 1.0 mM BEI at 37 °C. There was no antigen loss due to BEI treatment, and only a decrease in antigen levels was observed during storage. The sera from pigs immunized with antigen derived from a bioreactor exhibited a neutralizing antibody titer of approximately 1/1000 against Asia1 Shamir and Asia1/MOG/05 viruses; therefore, Asia1 Shamir-R is expected to provide sufficient protection against both viruses. If an FMD vaccine production facility is established, this Asia1 Shamir-R can be employed for domestic antigen banks in South Korea.

Africa (COVID-19) Vaccine Technology Transfer: Where Are We?

The rampant spread of the COVID-19 infection poses a grave and formidable challenge to global healthcare, with particular concern to the inhabitants of the African continent. In response to these pressing concerns, different strategies have been employed to combat the emergence of this insidious disease, encompassing crucial measures such as physical distancing, the utilization of face masks, meticulous hand hygiene, and widespread vaccination campaigns. Nevertheless, the economic realities faced by numerous African nations, characterized by their classification as "low-income countries (LICs)", present a formidable barrier to accessing and distributing approved vaccines to their populations. Moreover, it is essential to discuss the hesitancy of the European Union (EU) in releasing intellectual property rights associated with the transfer of vaccine technology to Africa. While the EU has been a key player in global efforts to combat the pandemic, there has been reluctance in sharing valuable knowledge and resources with African countries. This hesitancy raises concerns about equitable vaccine access and the potential for a prolonged health crisis in Africa. This review underscores the urgent imperative and need of establishing localized vaccine development and production facilities within Africa, necessitating the active involvement of governments and collaborative partnerships to achieve this crucial objective. Furthermore, this review advocates for the exploration of viable avenues for the transfer of vaccine technology as a means to facilitate equitable vaccine access across the African continent and also the cruciality and the need for the EU to reconsider its stance and actively engage in transferring vaccine technology to Africa through sharing intellectual property. The EU can contribute to the establishment of localized vaccine production facilities on the continent, which will not only increase vaccine availability but also promote self-sufficiency and resilience in the face of future health emergencies.

The effect of in vitro consecutive passages and culture medium on the genetic variations in BCG Pasteur 1173P2 vaccine.

Since the introduction of the Bacillus Calmette-Guérin (BCG) vaccine, the genomes of vaccine strains have undergone variations due to repeated passages in different laboratories and vaccine production facilities. Genetic variations have been considered as one of the effective factors in the BCG variable protective efficacy. Consecutive subcultures have been shown to play an essential role in causing genetic variations in several microorganisms, including Mycobacterium bovis BCG. Therefore, the world health organization (WHO) recommendation to limit the passages of master seed lot in the BCG vaccine production should be considered. Besides, the role of other external variables such as quality of the raw ingredients of the culture media, the type of the culture medium and the cultivation methods in the vaccine production has been poorly studied. Here, the effect of passages and culture medium on genetic variations in a BCG seed lot was investigated during a year. The findings of this study revealed a total of 19 variants compared to seed lot while the passages were more than the number recommended by WHO. The first culture of seed lot in the Sauton broth and Middlebrook 7H9 media, and the last subculture in Sauton broth had the least and the most variants, respectively. The observation of the higher number of variants in the last cultures on Sauton broth and Middlebrook 7H9 in comparison to the first and the middle cultures may indicate the effect of passages on the genetic variations in BCG. Additionally, more variants in BCG grown in the Sauton broth do not necessarily represent the greater ability of this medium to cause genetic mutations. For a better conclusion, it is required to examine the medium components as independent variables.

Implementing High-Reliability Organization Principles at Biological Diagnostic Laboratories: Case Study at National Institute of Health, Islamabad.

Healthcare organizations are complex systems where healthcare professionals, patients, biological materials, and equipment constantly interact and provide feedback with highly consequential outcomes. These are the characteristics of a complex adaptive system. Healthcare delivery requires coordination but it necessarily relies on delegation of essential functions. It is thus essential to have an engaged workforce to ensure optimal outcomes for patients. Thus human performance factors play a key role in ensuring both the presence of excellent healthcare provision and the absence of outcomes that must be avoided-"never events." The commitment of management was a precondition for the implementation of the high-reliability organization (HRO) principles. A team from middle management was engaged and provided with appropriate management tools for identifying, prioritizing, assessing, and applying solutions for the safety concern in their operating systems. This article documents efforts at the National Institute of Health (NIH) to adapt the principles of HROs to diagnostic laboratories and vaccine production facilities at its campus in Islamabad, Pakistan, and seeks to draw some lessons for how this approach can be usefully replicated in such facilities elsewhere. Public health institutes such as NIH deliver vital products and services that are inherently risky to produce, where the consequence of failure can be catastrophic. Adopting the HRO principles is an approach to improving not just safety, but also the overall organizational performance in any setting, including low-resource settings, and can serve as an implementable process for other institutions.

A Contamination Control Strategy Gone Wrong

Warning letters and regulatory inspection observation reports (e.g., FDA 483) often provide useful information for assessing risks in your facility and preparing for upcoming inspections. Starting with the updates to the European Union’s Annex 1 for the Manufacture of Sterile Drugs there has been an increased focus on contamination control strategies in facilities. A contamination control strategy is an integral part of pharmaceutical manufacturing, whether sterile or non-sterile. For this article we are going to look at a series of observations for a vaccine production facility and how we might learn from these observations. Highlighted are some of the contamination control issues. The FDA 483 Report was published in redacted form.

Effect of Extended Transit and Temperature Variations on EID50 Values of Thermostable NDVI-2 Vaccines

Temperature alterations due to poor cold keeping condition have consistently posed a challenge to vaccines dose EID50 titre value. The sensitivity of Newcastle disease vaccine to temperature variations due to poor cold storage condition in rural settings, led to the development of the NDVI-2 thermostable vaccine, which is known to exhibit a thermostability of 3 hours at 56 °C. In this study, three Newcastle disease vaccines (NDVI-2) with EID50 titre per dose values of log10 EID50 7.7, 7.6 and 6.75 respectively from vaccine production facility at the National Veterinary Research Institute (NVRI), Vom Nigeria, were sent for certification at African Union laboratory at Debre Zeit, Ethiopia under cold condition; using ice packs for cold chain maintenance. Tracking of the vaccine in the course of freight to the certifying laboratory revealed changes in the original transit route by the airline and delay by custom at the receiving country port. Thus, a 12 hours transit freight was delayed for over 7 days before arrival at the certifying laboratory. Though, the EID50 titre per dose values of these vaccines were originally well above recommended EID50 5.5 per dose value by log 2.2, log 2.1 and log 1.25 for NDVI-2 respectively. Comparing these vaccines EID50 values obtained from the certifying laboratory in relation to values initially obtained by the Quality Control Division of the producing Institute (NVRI), revealed loss of EID50 titre per dose value by log101.2, and log10 0.8 among two of the certified vaccine batches; with one of the certified vaccines retaining its original EID50 titre per dose value. This result attest that NDVI-2 thermostable vaccine produced at NVRI, Vom Nigeria are robust and of high quality. Thus, the findings of this study have further demonstrated that temperature variations, prolonged vaccine transportation under poor cold chain can affect the EID50 per dose value of a wholesome, field fit vaccine irrespective of the vaccine thermostability.

Maintaining excellence in a high volume academic cell therapy production facility through optimization of space utilization, operations, scheduling and staffing

Background & Aim The Clinical Cell and Vaccine Production Facility (CVPF) at UPenn is a FACT accredited resource with a 25 year record of novel first-in-human cell and gene therapy clinical trials. CVPF is integrated with research, process development, and correlative studies laboratories, QC/QA, clinical operations and monitoring, apheresis facilities and infusion sites. CVPF has two clean room facilities in which cell immunotherapy products are currently manufactured in support of >15 investigational new drug (IND) protocols. Methods, Results & Conclusion CVPF recently developed a clean room space utilization model to increase production capacity. Previously, a product occupied a single processing suite for the length of manufacture. Improvements to the processing room layout and implementation of a “bioreactor nursery” allowed for expansion of multiple products in a single space. Other benefits included fewer equipment pieces required and built-in redundancy. Additionally, we developed a standardized production slot model to reduce the rate of unfilled manufacturing slots and maximize facility efficiency. A standardized schedule along with the application of scheduling software has improved resource management, effort tracking, production task scheduling and capacity projections. Standardized scheduling is more efficient in scheduling manufacturing staff, sample transport, environmental monitoring, facility cleaning, equipment maintenance, QC testing, batch record preparation and review and product release. We developed an accelerated training program that reduced time-to-proficiency (TTP) for new hires through resource assignment, dedicated training space and re-designed curricula. A mock processing suite was established outside of the clean room space to allow for non-GMP materials to be used during training, greatly reducing cost. Still, staff recruitment, retention and training continue to be a challenge for academic manufacturing facilities as staff renewal limits production capacity and results in fewer patients enrolled in clinical trials. CVPF joined forces with University leadership to develop a novel metrics based career advancement track for cell therapy technologists. These techniques for increased productivity coupled with improved training methods and staff retention initiatives can be adopted by academic cell therapy facilities to optimize staffing, reduce costs and increase efficiencies, thereby increasing production capacity and patient clinical trial enrollment.

Early administration of tocilizumab (Toci) for the prevention of grade 4 cytokine release syndrome (CRS) after CD19-directed CAR T-cell therapy (CTL019)

Background & Aim The most common side effect of CD19-directed CAR-T cell therapy is CRS. IL-6 blockade by toci typically results in rapid resolution of severe CRS in CTL019 treated pts; however, the ideal timing of toci remains unclear. Pts with high tumor burden (TB; defined as ≥40% marrow blasts) are at high risk for severe CRS. We hypothesized that preemptive toci administration early in the course of CRS in pts with high TB might decrease acute CRS severity without compromising efficacy. Methods, Results & Conclusion MTD We conducted a two-cohort, open-label study of the timing of toci on CTL019-associated CRS. Pts were 1-24 yo with CD19-expressing relapsed or refractory B-cell acute lymphoblastic leukemia (r/r B-ALL). CTL019 was manufactured at the UPenn Cell and Vaccine Production Facility. Enrolled pts were assigned to high (≥ 40%) or low ( 38.5 C measured twice in 24 hours (“early toci”) and subsequently treated per standard CRS algorithm. Primary endpoint: frequency of gr4 CRS (Penn scale) in ≤5/15 pts. Secondary endpoints: D28 tumor response, remission duration, additional toxicities. RES 78 pts were screened, all were enrolled. 65 pts were evaluable for analysis at data cutoff. 15 pts were assigned to HTBC, 50 pts to LTBC and all were infused with CTL019 (median dose 5.0 × 106 [range 5.6 × 105-3.9 × 107] CAR+ T cells/kg). The overall rate of gr4 CRS was 9% (27% in the HTBC, 4% in the LTBC, Table 1). The overall rate of gr1-4 CRS was 72% (100% in the HTBC, 64% in the LTBC). One pt in the HTBC died from CNS hemorrhage after gr4 CRS. All patients in the HTBC received the early toci intervention. The overall rate of gr3 and 4 CRS was lower than in two prior studies of CTL019 therapy in pediatric r/r B-ALL (18% vs 42% in CHP959 and 48% in ELIANA). The ORR at day 28 follow-up was 97% (87% HTBC, 100% LTBC, with 71% CR and 26% CRi, Table 2). CONC We defined clincially meaningful reduction in severe CRS if ≤5/15 subjects had gr4 CRS in the HTBC; this a priori defined primary end-point was met. Early administration of toci targeted to pts with high TB may reduce the incidence of severe CRS while maintaining excellent efficacy of CTL019 therapy. TB remains a major predictor of toxicity. Longer follow-up is ongoing to assess impact on duration of remission.

Bovine monocyte derived dendritic cell based assay for measuring vaccine immunogenicity in vitro

During both human and animal vaccine development phases, animal testing is necessary to demonstrate vaccine efficacy. Since the number of antigen candidates for testing is usually large when developing a potential vaccine, it is too costly, time consuming and would involve higher risks to carry out selection using in vivo models. The currently available in vitro assays that measure immunogenicity do not adequately reproduce the in vivo state and this is especially true for vaccine research in livestock species. With this in mind, we have developed a bovine monocyte derived dendritic cell (MODC)s based assay to prime CD4 and CD8 lymphocytes in order to investigate vaccine immunogenicity in vitro. MODCs were generated, pulsed with diphtheria toxoid (DT) and co-cultured with lymphocytes for priming. Immunogenicity was measured through antigen recall when antigen pulsed MODC were re-introduced to the co-culture and proliferation of CD4 and CD8 positive lymphocytes were quantified using expressed Ki-67. Having developed the protocol for the assay, we then employed two licenced vaccines against blue tongue virus and rabies virus to validate the assay. Our results show the ability of the assay to satisfactorily measure immunogenicity in cattle. The assay could be used to identify antigens that induce CD4 and CD8 T cell responses prior to embarking on in vivo experiments and can also be used for the quality control of established vaccines in vaccine production facilities as a supplement for in vivo experiments.

Filovirus Research: How it Began.

The first reported filovirus outbreak occurred in August 1967, when laboratory workers in Marburg and Frankfurt, Germany, and Belgrade, Yugoslavia (now Serbia) became infected with an unknown highly pathogenic agent. The disease was characterized by high fever, malaise, rash, hemorrhagic and tetanic manifestations, and high lethality, amounting to 25%. The disease was introduced to Europe by grivets (Chlorocebus aethiops), which were used for biomedical research and vaccine production. The causative agent, Marburg virus, was isolated and identified by scientists of the University of Marburg, Germany in cooperation with specialists for viral electron microscopy at the Bernhard Nocht Institute in Hamburg, Germany. In this chapter, Dr. Slenczka, who was involved in the first isolation of Marburg virus in 1967, describes the desperate hunt of the causative agent of this first filovirus disease outbreak in the center of Europe, its successful isolation, the likely route of transmission from a monkey trading station to vaccine production facilities in Germany and Yugoslavia, and the consequences of this outbreak, including a shortage in the production of poliomyelitis vaccine In addition, this chapter provides insight into some of the peculiarities of filovirus infection, such as sexual virus transmission several months after recovery and the role of Ca2+-loss in Marburg virus pathogenesis, which were already observed during this first well-documented Marburg virus disease outbreak.

Challenges and successes for the grantees and the Technical Advisory Group of WHO’s influenza vaccine technology transfer initiative

One of the aims of the WHO Global Action Plan for Influenza Vaccines (GAP) was to transfer influenza vaccine production technology to interested manufacturers and governments in developing countries, to enable greater influenza vaccine manufacturing capacity against any pandemic threat or pandemic. For this objective, the GAP was supported by an independent Technical Advisory Group (TAG) to assist WHO to select vaccine manufacturing proposals for funding and to provide programmatic support for successful grantees. While there were many challenges, for both the TAG and grantees, there were also notable successes with an additional capacity of 338–600 million pandemic vaccine doses being made possible by the programme between 2007 and 2015, and a potential capacity of more than 600 million by 2016/17 with up to one billion doses expected by 2018/19. Seasonal vaccine production was also developed in 4 countries with another 4–5 countries expected to be producing seasonal vaccine by 2018/19. The relatively small WHO investments – in time and funding – made in these companies to develop their own influenza vaccine production facilities have had quite dramatic results.

Risk assessment, risk management and risk-based monitoring following a reported accidental release of poliovirus in Belgium, September to November 2014.

On 6 September 2014, the accidental release of 10(13) infectious wild poliovirus type 3 (WPV3) particles by a vaccine production plant in Belgium was reported. WPV3 was released into the sewage system and discharged directly to a wastewater treatment plant (WWTP) and subsequently into rivers that flowed to the Western Scheldt and the North Sea. No poliovirus was detected in samples from the WWTP, surface waters, mussels or sewage from the Netherlands. Quantitative microbial risk assessment (QMRA) showed that the infection risks resulting from swimming in Belgium waters were above 50% for several days and that the infection risk by consuming shellfish harvested in the eastern part of the Western Scheldt warranted a shellfish cooking advice. We conclude that the reported release of WPV3 has neither resulted in detectable levels of poliovirus in any of the samples nor in poliovirus circulation in the Netherlands. This QMRA showed that relevant data on water flows were not readily available and that prior assumptions on dilution factors were overestimated. A QMRA should have been performed by all vaccine production facilities before starting up large-scale culture of WPV to be able to implement effective interventions when an accident happens.

World Health Organization Guidelines for Containment of Poliovirus Following Type-Specific Polio Eradication - Worldwide, 2015.

In 1988, the World Health Assembly of the World Health Organization (WHO) resolved to eradicate polio worldwide. Among the three wild poliovirus (WPV) types (type 1, type 2, and type 3), WPV type 2 (WPV2) has been eliminated in the wild since 1999, and WPV type 3 (WPV3) has not been reported since 2012. In 2015, only Afghanistan and Pakistan have reported WPV transmission. On May 25, 2015, all WHO Member States endorsed World Health Assembly resolution 68.3 on full implementation of the Polio Eradication and Endgame Strategic Plan 2013-2018 (the Endgame Plan), and with it, the third Global Action Plan to minimize poliovirus facility-associated risk (GAPIII). All WHO Member States have committed to implementing appropriate containment of WPV2 in essential laboratory and vaccine production facilities* by the end of 2015 and of type 2 oral poliovirus vaccine (OPV2) within 3 months of global withdrawal of OPV2, which is planned for April 2016. This report summarizes critical steps for essential laboratory and vaccine production facilities that intend to retain materials confirmed to contain or potentially containing type-specific WPV, vaccine-derived poliovirus (VDPV), or OPV/Sabin viruses, and steps for nonessential facilities† that process specimens that contain or might contain polioviruses. National authorities will need to certify that the essential facilities they host meet the containment requirements described in GAPIII. After certification of WPV eradication, the use of all OPV will cease; final containment of all polioviruses after polio eradication and OPV cessation will minimize the risk for reintroduction of poliovirus into a polio-free world.

Comprehensive hands-on training for influenza vaccine manufacturing: a WHO-BARDA-BTEC partnership for global workforce development.

The critical need for enhancing influenza pandemic preparedness in many developing nations has led the World Health Organization (WHO) and the Biomedical Advanced Research and Development Authority (BARDA), part of the U.S. Department of Health and Human Services (HHS), to develop an international influenza vaccine capacity-building program. Among the critical limitations faced by many of these nations is lack of access to training programs for staff supporting operations within vaccine production facilities. With support from BARDA, the Biomanufacturing Training and Education Center (BTEC) at North Carolina State University has addressed this need for training by developing and delivering a comprehensive training program, consisting of three courses: Fundamentals of cGMP Influenza Vaccine Manufacturing, Advanced Upstream Processes for Influenza Vaccine Manufacturing, and Advanced Downstream Processes for Influenza Vaccine Manufacturing. The courses cover process design, transfer, and execution at manufacturing scale, quality systems, and regulations covering both manufacturing and approval of pandemic vaccines. The Fundamentals course focuses on the concepts, equipment, applicable regulations, and procedures commonly used to produce influenza vaccine. The two Advanced courses focus on process design, scale up, validation, and new technologies likely to improve efficiency of vaccine production. All three courses rely on a combination of classroom instruction and hands-on training in BTEC's various laboratories. Each course stands alone, and participants may take one or more of the three courses. Overall participant satisfaction with the courses has been high, and follow-up surveys show that participants actively transferred the knowledge they gained to the workplace. Future plans call for BTEC to continue offering the three courses and to create an online version of several modules of the Fundamentals course.

English

The appropriate use of biocides is essential in any vaccine production facility and their proper evaluation using standardized tests marks the first step to ensure their proper use. Quantitative suspension tests against reference and environmental isolates were carried out to evaluate the efficacy of various biocides used in the main vaccine production facility in Egypt. Several use-dilutions of the biocides were evaluated at contact times of up to 5 min in case of antiseptics and 30 min in case of disinfectants to measure the biocide activity against bacteria, fungi and spores. Standard strains were used in addition to the main bacterial isolates identified in an environmental monitoring program carried out in the same facility. Alcohol based biocides showed bactericidal and fungicidal activity but no sporicidal activity. Chlorine based compounds and glutaraldehyde showed bactericidal and fungicidal effect while, the sporicidal effects depended on the used dilution and the contact time. Hydrogen peroxide showed bactericidal, fungicidal and sporicidal activity. Quaternary ammonium compounds tested showed very weak activity in all tests. Evaluation of the biocide is an essential step to guarantee the use of the most appropriate agent in any location.   Key words: Biocides, antiseptics, disinfectants, vaccines, Egypt.

Efficacy and toxicity of neutralizers against disinfectants and antiseptics used in vaccine production facility

Effective neutralization of chemical biocide is the first step in the accurate evaluation of antiseptics and disinfectants to avoid overestimation of the biocide activity. The aim of this study is to screen novel neutralizers (or neutralizer combinations) against different antiseptics and disinfectants that are highly used in the principle vaccine production facility in Egypt. Neutralizer efficacy (NE) ratios were determined by comparing the recovery of the challenging index microorganisms from the neutralizing solution in the presence, or the absence, of the biocide. Neutralizer toxicity (NT) ratios were determined by comparing the recovery of viable microorganisms in the neutralizer exposed population and the viability population. An effective and non-toxic neutralizer was initially identified by NE and NT ratios of ≥ 0.75 (75% recovery). The most potent neutralizers were 1% physiological peptone water, 0.1 % Tween 80 in 1% physiological peptone, 0.6% sodium thiosulphate and Dey-Engley. Due to the absence of a universal neutralizer, the evaluation of NE and toxicity is an essential step in testing any new biocide. The evaluation of neutralizer activity should be carried out separately for each index microorganism and neutralizer pair to identify the most potent compound in each case. Key words: Antiseptics, disinfectants, neutralizers.

WHO influenza vaccine technology transfer initiative: Role and activities of the Technical Advisory Group

In May 2006, the WHO published a Global Pandemic Influenza Action Plan. A significant part of that plan involves the transfer of technology necessary to build production capacity in developing countries. The WHO influenza technology transfer initiative has been successful. Clearly the relatively small WHO investments made in these companies to develop their own influenza vaccine production facilities have had quite dramatic results. A few companies are already producing large amounts of influenza vaccine. Others will soon follow. Whether they are developing egg-based or planning non-egg based influenza vaccine production, all companies are optimistic that their efforts will come to fruition.

Foot-and-mouth disease vaccine potency testing: The influence of serotype, type of adjuvant, valency, fractionation method, and virus culture on the dose–response curve in cattle

The aim of this study was to determine a relationship between vaccine potency (amount of PD 50 per dose) and fraction of clinically protected cattle following homologous challenge with infectious foot-and-mouth disease (FMD) virus, and to determine the effect of method of fractionation, serotype, type of adjuvant, valency and type of virus culture on the dose–response curve. Data from 297 potency tests of FMD vaccines, comprising 4004 vaccinated cattle, performed at the FMD vaccine production facility in the Netherlands, were used for the present study. A generalised linear mixed effect model was used to analyse the results. Our study showed that the relation between FMD vaccine potency and fraction protected was also affected by the serotype and type of adjuvant. No common level of protection could be assigned to all FMD vaccines with the same amount of PD 50 per dose, this information is essential when designing a new standard FMD vaccines control.

Dendritic Cell Vaccine Production Facility: From Design to Operation

Dendritic Cell Vaccine Production Facility: From Design to Operation

La experimentación con animales en relación con el descubrimiento y la producción de vacunas de uso veterinario

Veterinary vaccine research, development and production facilities must aim to improve animal welfare, respond to public concerns and meet regulatory requirements, while at the same time fulfilling their objective of producing evermore effective and safer vaccines. The use of animal experimentation for the development of new veterinary vaccines is inevitable, as no in vitro model can predict a candidate vaccine's ability to induce protection in the target species. Against the backdrop of ethical and regulatory constraints, constant progress is being made in creating the best possible conditions for animal experimentation. Keeping up to date with the constant changes in the field of animal ethics requires a particular effort on the part of the pharmaceutical industry, which must make careful changes to product registration documentation in accordance with each new development.