Following a request from the European Commission, the EFSA Panel on Animal Health and Welfare (AHAW) was asked to deliver a scientific opinion on the use of a low atmosphere pressure system (LAPS) for stunning poultry prior to slaughter. Four studies were submitted for assessment. The LAPS method aims to render poultry unconscious by gradually reducing oxygen levels in the atmosphere. For a new method to be allowed in the EU, it must ensure animal welfare equivalent to currently permitted methods.
The AHAW Panel applied eligibility criteria from its guidance document to assess whether the key parameters around the LAPS intervention and animal welfare outcomes were described adequately. It found that none of the submitted studies provided enough information on
EFSA Guidance on assessment new stunning & slaughter methodsHarm Kiezebrink
The guidance document defines the assessment process and criteria that will be applied by EFSA's Animal Health and Welfare Panel to evaluate studies on new or modified legal stunning interventions for slaughter animals. Studies must fulfill eligibility criteria based on legislation and scientific evidence, as well as reporting quality and methodological quality criteria.
The eligibility criteria focus on the intervention and required outcomes of immediate unconsciousness/insensibility, absence of avoidable suffering, and duration of unconsciousness until death. Studies meeting eligibility criteria will be further assessed based on adherence to reporting guidelines and methodological rigor.
If criteria are fulfilled, a full assessment of animal welfare implications and evidence quality will be conducted. If not, shortcomings will be identified to guide improvements
This document summarizes the outcome of a public consultation on a draft guidance document from the European Food Safety Authority (EFSA) regarding assessment criteria for studies evaluating the effectiveness of stunning interventions for animal protection during slaughter. EFSA received comments from six parties and has assessed and addressed the comments in finalizing the guidance document. The key comments received concerned specifying additional parameters for characterizing stunning interventions and clarifying certain aspects of the guidance. EFSA and its Animal Health and Welfare Panel have updated the guidance document based on the comments received to improve its comprehensiveness and clarity.
EFSA paper on monitoring procedures at slaughterhousesHarm Kiezebrink
The objective of this review was to summarize the currently available data describing the sensitivity and specificity of indicators of unconsciousness and death in the following stun-kill methods and species combinations:
1) Penetrative captive bolt for bovine animals
2) Head-only electrical stunning for pigs
3) Head-only electrical stunning for sheep and goats
4) Electrical waterbath for poultry (chickens and turkeys)
5) Carbon dioxide at high concentration for pigs
6) All authorized gas methods to slaughter chickens and turkeys (carbon dioxide at high concentration, carbon dioxide in two phases, carbon dioxide associated with inert gases and inert gases alone)
7) Slaughter without stunning for bovine animals
8) Slaughter without stunning for sheep and goats
9) Slaughter without stunning for chickens and turkeys
The reference tests for unconsciousness and death were to have been measured using electroencephalography (EEG). The definition of unconsciousness and death based on EEG were not specified, and the definition used by authors was reported. The index tests of interest were a variety of indicators requested by the funding agency such as no corneal reflex and immediate collapse.
The index tests differed by stun-kill methods and species combination. A comprehensive search identified 22 publications contained 24 species-stun/kill method combinations.
No studies explicitly reported the sensitivity and specificity of the indicators in conscious and unconscious animals. Many studies reported the proportion of stunned animals with indicators, rather than the proportion of unconscious or conscious animals at a set time point with the indicators. Such data could not be translated into sensitivity and specificity.
Other studies reported the average time to occurrence of an indicator or average time to cessation of the indicators. Such data cannot be translated into sensitivity and specificity estimates without knowledge of the joint distributions.
Gas alternatives to carbon dioxide for euthanasia a piglet perspectiveHarm Kiezebrink
The use of nitrous oxide as an anesthetic/euthanasia agent may prove to be affordable, feasible and more humane than other alternatives.
The neonatal stage is a critical time in the life of a pig, when they are prone to become sick or weak. This is the stage at which most euthanasia procedures are required if the pig is judged unable to recover. Any euthanasia method should be humane, practical, economical and socially acceptable to be universally accepted.
They found that nitrous oxide in oxygen appeared to be less aversive than nitrous oxide, nitrogen, or argon all combined with low (30%) concentrations of carbon dioxide or 90% carbon dioxide by itself.
This study is the first to investigate the use of nitrous oxide at sufficiently high concentrations to cause anesthesia. Nitrous oxide, commonly referred to as laughing gas, has been widely used in human surgery and dental offices for its pain-relieving, sedative and anxiolytic effects. It is cheap, non-flammable, non-explosive, legally accessible and not classified as a drug in the U.S., and already commonly used in the food industry as a propellant for food products.
Development of its use into an automated procedure will allow producers to implement it with little effort. Thus its use as an anesthetic/euthanasia agent may prove to be affordable, feasible and more humane than other alternatives.
Rabbit farming is a small-scale industry that does not have a major national or international representative organisation in most of the EU countries. Over 76% of the total production in the EU is in Italy, Spain and France, and home production is still widespread. The production of jointed and processed products is increasing rapidly compared with whole carcase sales and rabbit meat consumption, although less than other meats, is still significant in some countries.
Ethical issues in animal experimentation (with emphasis on CPCSEA guidelines)Sandeep Lahiry
The document discusses ethical issues related to animal experimentation, summarizing Indian and international guidelines. It outlines the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA) which regulates animal testing in India according to the Prevention of Cruelty to Animals Act. The CPCSEA guidelines emphasize minimizing animal use, pain and suffering, and proper housing and care. Institutional Animal Ethics Committees must approve all research projects involving animals.
This systematic review evaluated the effects of glucocorticoid with cyclophosphamide treatment for paraquat-induced lung fibrosis based on three randomized controlled trials with a total of 164 participants. The review found that patients receiving glucocorticoid with cyclophosphamide in addition to standard care had a lower risk of death at final follow-up compared to those receiving standard care alone, with a risk ratio of 0.72 (95% CI 0.59 to 0.89). Based on these findings, the authors concluded that glucocorticoid with cyclophosphamide may provide a beneficial treatment for patients with paraquat-induced lung fibrosis. However, they noted that the studies were small and one was of low quality, so the benefits need
This was my first podium presentation presented at an international conference organized by UNESCO. The conference was remarkable because it involved superspecialty field to even nursing staff. My presentation was amongst the contenders for prize distribution. However, it did not happen so due to other presenters who outperformed me.
EFSA Guidance on assessment new stunning & slaughter methodsHarm Kiezebrink
The guidance document defines the assessment process and criteria that will be applied by EFSA's Animal Health and Welfare Panel to evaluate studies on new or modified legal stunning interventions for slaughter animals. Studies must fulfill eligibility criteria based on legislation and scientific evidence, as well as reporting quality and methodological quality criteria.
The eligibility criteria focus on the intervention and required outcomes of immediate unconsciousness/insensibility, absence of avoidable suffering, and duration of unconsciousness until death. Studies meeting eligibility criteria will be further assessed based on adherence to reporting guidelines and methodological rigor.
If criteria are fulfilled, a full assessment of animal welfare implications and evidence quality will be conducted. If not, shortcomings will be identified to guide improvements
This document summarizes the outcome of a public consultation on a draft guidance document from the European Food Safety Authority (EFSA) regarding assessment criteria for studies evaluating the effectiveness of stunning interventions for animal protection during slaughter. EFSA received comments from six parties and has assessed and addressed the comments in finalizing the guidance document. The key comments received concerned specifying additional parameters for characterizing stunning interventions and clarifying certain aspects of the guidance. EFSA and its Animal Health and Welfare Panel have updated the guidance document based on the comments received to improve its comprehensiveness and clarity.
EFSA paper on monitoring procedures at slaughterhousesHarm Kiezebrink
The objective of this review was to summarize the currently available data describing the sensitivity and specificity of indicators of unconsciousness and death in the following stun-kill methods and species combinations:
1) Penetrative captive bolt for bovine animals
2) Head-only electrical stunning for pigs
3) Head-only electrical stunning for sheep and goats
4) Electrical waterbath for poultry (chickens and turkeys)
5) Carbon dioxide at high concentration for pigs
6) All authorized gas methods to slaughter chickens and turkeys (carbon dioxide at high concentration, carbon dioxide in two phases, carbon dioxide associated with inert gases and inert gases alone)
7) Slaughter without stunning for bovine animals
8) Slaughter without stunning for sheep and goats
9) Slaughter without stunning for chickens and turkeys
The reference tests for unconsciousness and death were to have been measured using electroencephalography (EEG). The definition of unconsciousness and death based on EEG were not specified, and the definition used by authors was reported. The index tests of interest were a variety of indicators requested by the funding agency such as no corneal reflex and immediate collapse.
The index tests differed by stun-kill methods and species combination. A comprehensive search identified 22 publications contained 24 species-stun/kill method combinations.
No studies explicitly reported the sensitivity and specificity of the indicators in conscious and unconscious animals. Many studies reported the proportion of stunned animals with indicators, rather than the proportion of unconscious or conscious animals at a set time point with the indicators. Such data could not be translated into sensitivity and specificity.
Other studies reported the average time to occurrence of an indicator or average time to cessation of the indicators. Such data cannot be translated into sensitivity and specificity estimates without knowledge of the joint distributions.
Gas alternatives to carbon dioxide for euthanasia a piglet perspectiveHarm Kiezebrink
The use of nitrous oxide as an anesthetic/euthanasia agent may prove to be affordable, feasible and more humane than other alternatives.
The neonatal stage is a critical time in the life of a pig, when they are prone to become sick or weak. This is the stage at which most euthanasia procedures are required if the pig is judged unable to recover. Any euthanasia method should be humane, practical, economical and socially acceptable to be universally accepted.
They found that nitrous oxide in oxygen appeared to be less aversive than nitrous oxide, nitrogen, or argon all combined with low (30%) concentrations of carbon dioxide or 90% carbon dioxide by itself.
This study is the first to investigate the use of nitrous oxide at sufficiently high concentrations to cause anesthesia. Nitrous oxide, commonly referred to as laughing gas, has been widely used in human surgery and dental offices for its pain-relieving, sedative and anxiolytic effects. It is cheap, non-flammable, non-explosive, legally accessible and not classified as a drug in the U.S., and already commonly used in the food industry as a propellant for food products.
Development of its use into an automated procedure will allow producers to implement it with little effort. Thus its use as an anesthetic/euthanasia agent may prove to be affordable, feasible and more humane than other alternatives.
Rabbit farming is a small-scale industry that does not have a major national or international representative organisation in most of the EU countries. Over 76% of the total production in the EU is in Italy, Spain and France, and home production is still widespread. The production of jointed and processed products is increasing rapidly compared with whole carcase sales and rabbit meat consumption, although less than other meats, is still significant in some countries.
Ethical issues in animal experimentation (with emphasis on CPCSEA guidelines)Sandeep Lahiry
The document discusses ethical issues related to animal experimentation, summarizing Indian and international guidelines. It outlines the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA) which regulates animal testing in India according to the Prevention of Cruelty to Animals Act. The CPCSEA guidelines emphasize minimizing animal use, pain and suffering, and proper housing and care. Institutional Animal Ethics Committees must approve all research projects involving animals.
This systematic review evaluated the effects of glucocorticoid with cyclophosphamide treatment for paraquat-induced lung fibrosis based on three randomized controlled trials with a total of 164 participants. The review found that patients receiving glucocorticoid with cyclophosphamide in addition to standard care had a lower risk of death at final follow-up compared to those receiving standard care alone, with a risk ratio of 0.72 (95% CI 0.59 to 0.89). Based on these findings, the authors concluded that glucocorticoid with cyclophosphamide may provide a beneficial treatment for patients with paraquat-induced lung fibrosis. However, they noted that the studies were small and one was of low quality, so the benefits need
This was my first podium presentation presented at an international conference organized by UNESCO. The conference was remarkable because it involved superspecialty field to even nursing staff. My presentation was amongst the contenders for prize distribution. However, it did not happen so due to other presenters who outperformed me.
The document discusses principles and guidelines regarding the use of animals in research experiments. It summarizes key points from international guidelines developed in 1985 by CIOMS and endorsed by WHO and EMRC. The principles emphasize replacing animal experiments where possible, using the minimum number of animals required, avoiding pain and distress, proper housing and care of animals, and ensuring researchers are qualified to conduct procedures on animals. It also discusses investigator responsibilities to apply the Three Rs (Replacement, Reduction and Refinement) and keep detailed records of animal experiments.
Acute, subacute, and chronic toxicity studies in animals are described. Various OECD test guidelines are provided for acute toxicity testing via oral, dermal, and inhalation routes. Repeated dose 28-day and 90-day oral and dermal toxicity studies in rodents are used to evaluate subacute and chronic effects. A variety of endpoints are evaluated including clinical signs, hematology, gross necropsy, and histopathology. Chronic toxicity studies aim to identify target organs, characterize dose-response relationships, and identify no-observed-adverse-effect levels. The prediction of human chronic toxicity from animal studies and hypotheses testing on modes of action are also objectives.
The document discusses OECD Guideline 423 for acute oral toxicity testing. It outlines that the guideline uses a stepwise testing process with only 3 animals per step to classify chemicals according to their toxicity while minimizing animal use. The testing involves observing animals for signs of toxicity after administering chemical doses and determining subsequent dose levels based on mortality responses. The goal is to classify chemicals according to the globally harmonized system to improve knowledge of hazardous chemicals.
Alternative methods to animal toxicity testingSachin Sharma
This document presents information on alternative methods to animal toxicity testing. It discusses the need for alternatives due to the harms animals face in toxicity testing. The 3Rs principles of reduction, refinement and replacement are explained, which aim to minimize animal use and suffering. The validation process for new alternative methods through organizations like ECVAM is covered. Specific alternative methods mentioned include in vitro tests like the Ames test and HET-CAM test, in silico methods, and mathematical models. The HET-CAM test for eye irritation is described in more detail.
Research Methods: Ethics II (Animal Research)Brian Piper
lecture 3 from a college level research methods in psychology course taught in the spring 2012 semester by Brian J. Piper, Ph.D. (psy391@gmail.com) at Linfield College, includes IACUC, animal welfare act, refinement, reduction, replacement
This document discusses preclinical toxicity studies, which are animal tests done before clinical trials in humans to evaluate the safety and biological effects of investigational drugs. It describes the goals of acute, sub-acute, and chronic toxicity tests in rodents like rats and non-rodents like dogs to determine pharmacokinetics, pharmacodynamics, and toxicity. The choice of animal species and use of two species is aimed to extrapolate potential drug effects in humans. The document also discusses how toxicity test data is used to calculate therapeutic indices and estimate safe starting doses for clinical trials.
Subacute toxicity testing is conducted over 28 days to estimate safety margins of substances. It involves exposing test animals like rats to substances via oral, dermal or inhalation routes and observing for signs of toxicity. Key guidelines for subacute toxicity testing are OECD 407 for oral exposure, OECD 410 for dermal exposure, and OECD 412 for inhalation exposure. These studies provide important toxicity data used in chemical risk assessments.
The document describes guidelines for toxicological screening methods from the Organisation for Economic Co-Operation and Development (OECD). It outlines various OECD guidelines for toxicity studies including acute oral toxicity, acute dermal toxicity, skin sensitization, acute eye irritation, inhalation toxicity, reproduction/developmental toxicity screening, carcinogenicity studies, prenatal developmental toxicity studies, and genetic toxicology studies. It then describes the methods, measurements, time course studies, and data reporting for conducting toxicological screening studies.
Redbook 2000: General Guidelines for Designing and Conducting ToxicityStudies.Dmitri Popov
This document provides guidelines for designing and conducting toxicity studies, including recommendations for animal care, selection of species/strains, clinical testing procedures, urinalysis, neurotoxicity and immunotoxicity screening, and microscopic evaluation of tissues. It recommends following NIH and DHEW guidelines for animal care and housing. Both male and female rodents (usually rats) and non-rodents (usually dogs) should be used. Ophthalmological exams, hematology, clinical chemistry, and urinalysis should be conducted according to schedules in other FDA guidelines. Tissues should be examined microscopically for abnormalities.
The guidelines describe about the subacute toxicity studies in rodents with a comparison with the previous guideline.it also includes the comparison of all three subacute toxicity studies OECD 407, OECD 410, and OECD 412
OECD Guideline For Acute oral toxicity (TG 423)Naveen K L
This document provides guidelines for conducting acute oral toxicity tests using animals to determine the toxicological effects of substances. It describes the key aspects of the test including the principles, animal selection, dose preparation and administration, observation periods, and reporting of results. The goal is to classify substances based on the number of animals affected at different dose levels in a stepwise testing process using a minimum number of animals to obtain sufficient information on a substance's acute toxicity and enable its classification.
This document discusses experimenting on animals and outlines a work plan. It asks why we should experiment on animals and what the terms of any experiments should be. It then provides some examples of successful experiments including work on malaria, cancer, and drug development.
ICH guidelines provide standards for toxicity studies to ensure safe, effective, and high quality pharmaceutical products. Guideline S3A deals with conducting toxicity studies and quantifying exposure. General principles include quantifying exposure levels in different species and sexes using plasma concentration or area under the curve. Toxicokinetic studies should be performed to determine metabolite levels and justify dose levels. Reporting should include detailed toxicokinetic data and evaluation. Toxicokinetics are assessed in various toxicity studies including single dose studies, repeated dose studies, genotoxicity studies, carcinogenicity studies, and reproductive toxicity studies.
EFSA Opinion on electrical requirements for poultry waterbath stunning equipm...Harm Kiezebrink
In July 2014, EFSA provided her opinion on a study that proposes parameters for poultry electrical waterbath stunning different to those laid down in Council Regulation EU 1099/2009 on the protection of animals at the time of killing.
The submitted study reports upon the use (mean + SD) of a current of 104.00 ± 3.88 mA, a voltage of 125.86 ± 3.28 V and a frequency of 589.78 ± 0.63 Hz using a square wave in alternating current (AC) with a 50 % duty cycle. These conditions were applied for 15 seconds to chickens under laboratory and slaughterhouse conditions.
The methodology and the data reported do not provide conclusive evidence that the combination of the proposed electrical frequency and current induced unconsciousness without exposing the chickens to avoidable pain and suffering, and some chickens did not remain unconscious for a sufficient time to prevent avoidable pain and suffering during slaughter.
EFSA stated in their report that it was doubtful that recovery of consciousness could be avoided prior to neck cutting and/or during bleeding. The minimum duration of unconsciousness was reported to be 11 seconds, which is too short to permit a feasible stun-to-stick interval. Further, it is also doubtful that recovery of consciousness could be avoided prior to neck cutting and/or during bleeding. The minimum time to resumption of breathing was reported to be 8 seconds following stunning.
Because the information provided in the study was incomplete and insufficient, it did not pass the eligibility phase of the assessment. The information provided was sufficient to conclude that the birds were not rendered immediately unconscious by the intervention. Application of a current less than that required inducing immediate unconsciousness causes pain, distress and suffering. The study failed to demonstrate absence of pain and suffering until onset of unconsciousness. The minimum duration of unconsciousness was too short to ensure unconsciousness until death by bleeding.
Toxicity studies in animals are conducted to identify any toxic effects of a substance prior to clinical use in humans. The document outlines various types of toxicity studies including acute, subacute, chronic, and lethality studies. Acute studies involve a single high dose to determine toxic effects over 14 days, while repeated dose studies like subacute and chronic studies administer multiple lower doses over weeks to years to identify target organ toxicity. Lethality studies determine the lethal dose for 50% of animals (LD50). Systemic toxicity parameters evaluated include effects on liver, kidney, heart and other organs. Toxicity studies provide safety information required for approval to conduct human clinical trials.
The document summarizes guidelines for skin sensitization toxicity studies using guinea pigs. It describes two common test types - adjuvant tests using Freund's Complete Adjuvant and non-adjuvant tests like the Buehler test. The preferred methods are the Guinea Pig Maximization Test and the non-adjuvant Buehler Test. Both tests involve induction exposure, a rest period, then challenge exposure to determine skin reaction responses. Precise protocols for each test are provided regarding animal selection, housing, dosing, observations, scoring, and reporting of results.
Introductiontoexperimentalpharmacologicalresearch 211008134020PHARMA IQ EDUCATION
This document provides an introduction to experimental pharmacological research. It discusses the definitions of pharmacology and experimental pharmacology. Experimental pharmacology involves both pre-clinical and clinical studies and can lead to new therapeutic agents or identification of toxic effects. The objectives of experimental pharmacology are to provide scientific knowledge of drugs, discover new drugs and techniques. Experimentation can be done in vivo using animal models like rats, mice and rabbits, or in vitro using isolated tissues, cells, or cell-free components. Selection of animal models considers factors like size, availability, sensitivity and species-specific responses. In vivo and in vitro methods are vital experimental tools in pharmacological research.
EU REACH regulation changed the way to do chemical risk assessment. All chemicals marketed or manufactured in the EU must have its own dossier. Non standard methods including alternatives to animal testing are accepted.
Half Italian, half English
Acute and repeated dose toxicity studies are important for determining the safety of pharmaceuticals intended for human use. Key details include:
- Acute toxicity studies involve single high doses to determine the median lethal dose (LD50) and maximum tolerated dose. Repeated dose studies last 14 days to 3 months.
- Studies are conducted in at least two species, often rats and non-rodents, to identify target organs and toxicity reversibility.
- Animals are observed for signs of toxicity and mortality. Necropsies and histopathology are performed to identify organ damage.
- Sub-acute studies last 14-90 days to determine effects of repeated administration and establish doses for longer term studies.
EFSA AHAW report on monitoring procedures at poultry slaughterhousesHarm Kiezebrink
The EFSA Panel on Animal Health and Welfare (AHAW) was asked to deliver scientific opinions on monitoring procedures at slaughterhouses for different animal species, stunning methods and slaughter without stunning. AHAW agreed that, although it is traditional to look for outcomes of unconsciousness in poultry following stunning, the risk of poor welfare can be detected better if bird welfare monitoring is focused on detecting consciousness, i.e. ineffective stunning or recovery of consciousness.
Therefore, the indicators were phrased neutrally (e.g. corneal reflex) and the outcomes were phrased either suggesting unconsciousness (e.g. absence of corneal reflex) or suggesting consciousness (e.g. presence of corneal reflex). This approach is commonly used in animal health studies (e.g. testing for the presence of a disease) but very new to animal welfare monitoring in slaughterhouses.
A toolbox of selected indicators is proposed to check for signs of consciousness in poultry after stunning with waterbaths or gas mixtures; a different toolbox of indicators is proposed for confirming death of the birds following slaughter without stunning.
This study investigates the possibility of a harmonized EU approach to assessing the added therapeutic value (ATV) of medicines. It finds that while ATV is not systematically assessed for marketing authorization, most EU countries do consider it as part of pricing and reimbursement decisions. The study reviews the EU legal framework, explores ATV practices across the EU-28, and provides an in-depth analysis of ATV in six countries. It closes by outlining recommendations for a possible European harmonization of ATV assessment within the current legal framework.
The document discusses principles and guidelines regarding the use of animals in research experiments. It summarizes key points from international guidelines developed in 1985 by CIOMS and endorsed by WHO and EMRC. The principles emphasize replacing animal experiments where possible, using the minimum number of animals required, avoiding pain and distress, proper housing and care of animals, and ensuring researchers are qualified to conduct procedures on animals. It also discusses investigator responsibilities to apply the Three Rs (Replacement, Reduction and Refinement) and keep detailed records of animal experiments.
Acute, subacute, and chronic toxicity studies in animals are described. Various OECD test guidelines are provided for acute toxicity testing via oral, dermal, and inhalation routes. Repeated dose 28-day and 90-day oral and dermal toxicity studies in rodents are used to evaluate subacute and chronic effects. A variety of endpoints are evaluated including clinical signs, hematology, gross necropsy, and histopathology. Chronic toxicity studies aim to identify target organs, characterize dose-response relationships, and identify no-observed-adverse-effect levels. The prediction of human chronic toxicity from animal studies and hypotheses testing on modes of action are also objectives.
The document discusses OECD Guideline 423 for acute oral toxicity testing. It outlines that the guideline uses a stepwise testing process with only 3 animals per step to classify chemicals according to their toxicity while minimizing animal use. The testing involves observing animals for signs of toxicity after administering chemical doses and determining subsequent dose levels based on mortality responses. The goal is to classify chemicals according to the globally harmonized system to improve knowledge of hazardous chemicals.
Alternative methods to animal toxicity testingSachin Sharma
This document presents information on alternative methods to animal toxicity testing. It discusses the need for alternatives due to the harms animals face in toxicity testing. The 3Rs principles of reduction, refinement and replacement are explained, which aim to minimize animal use and suffering. The validation process for new alternative methods through organizations like ECVAM is covered. Specific alternative methods mentioned include in vitro tests like the Ames test and HET-CAM test, in silico methods, and mathematical models. The HET-CAM test for eye irritation is described in more detail.
Research Methods: Ethics II (Animal Research)Brian Piper
lecture 3 from a college level research methods in psychology course taught in the spring 2012 semester by Brian J. Piper, Ph.D. (psy391@gmail.com) at Linfield College, includes IACUC, animal welfare act, refinement, reduction, replacement
This document discusses preclinical toxicity studies, which are animal tests done before clinical trials in humans to evaluate the safety and biological effects of investigational drugs. It describes the goals of acute, sub-acute, and chronic toxicity tests in rodents like rats and non-rodents like dogs to determine pharmacokinetics, pharmacodynamics, and toxicity. The choice of animal species and use of two species is aimed to extrapolate potential drug effects in humans. The document also discusses how toxicity test data is used to calculate therapeutic indices and estimate safe starting doses for clinical trials.
Subacute toxicity testing is conducted over 28 days to estimate safety margins of substances. It involves exposing test animals like rats to substances via oral, dermal or inhalation routes and observing for signs of toxicity. Key guidelines for subacute toxicity testing are OECD 407 for oral exposure, OECD 410 for dermal exposure, and OECD 412 for inhalation exposure. These studies provide important toxicity data used in chemical risk assessments.
The document describes guidelines for toxicological screening methods from the Organisation for Economic Co-Operation and Development (OECD). It outlines various OECD guidelines for toxicity studies including acute oral toxicity, acute dermal toxicity, skin sensitization, acute eye irritation, inhalation toxicity, reproduction/developmental toxicity screening, carcinogenicity studies, prenatal developmental toxicity studies, and genetic toxicology studies. It then describes the methods, measurements, time course studies, and data reporting for conducting toxicological screening studies.
Redbook 2000: General Guidelines for Designing and Conducting ToxicityStudies.Dmitri Popov
This document provides guidelines for designing and conducting toxicity studies, including recommendations for animal care, selection of species/strains, clinical testing procedures, urinalysis, neurotoxicity and immunotoxicity screening, and microscopic evaluation of tissues. It recommends following NIH and DHEW guidelines for animal care and housing. Both male and female rodents (usually rats) and non-rodents (usually dogs) should be used. Ophthalmological exams, hematology, clinical chemistry, and urinalysis should be conducted according to schedules in other FDA guidelines. Tissues should be examined microscopically for abnormalities.
The guidelines describe about the subacute toxicity studies in rodents with a comparison with the previous guideline.it also includes the comparison of all three subacute toxicity studies OECD 407, OECD 410, and OECD 412
OECD Guideline For Acute oral toxicity (TG 423)Naveen K L
This document provides guidelines for conducting acute oral toxicity tests using animals to determine the toxicological effects of substances. It describes the key aspects of the test including the principles, animal selection, dose preparation and administration, observation periods, and reporting of results. The goal is to classify substances based on the number of animals affected at different dose levels in a stepwise testing process using a minimum number of animals to obtain sufficient information on a substance's acute toxicity and enable its classification.
This document discusses experimenting on animals and outlines a work plan. It asks why we should experiment on animals and what the terms of any experiments should be. It then provides some examples of successful experiments including work on malaria, cancer, and drug development.
ICH guidelines provide standards for toxicity studies to ensure safe, effective, and high quality pharmaceutical products. Guideline S3A deals with conducting toxicity studies and quantifying exposure. General principles include quantifying exposure levels in different species and sexes using plasma concentration or area under the curve. Toxicokinetic studies should be performed to determine metabolite levels and justify dose levels. Reporting should include detailed toxicokinetic data and evaluation. Toxicokinetics are assessed in various toxicity studies including single dose studies, repeated dose studies, genotoxicity studies, carcinogenicity studies, and reproductive toxicity studies.
EFSA Opinion on electrical requirements for poultry waterbath stunning equipm...Harm Kiezebrink
In July 2014, EFSA provided her opinion on a study that proposes parameters for poultry electrical waterbath stunning different to those laid down in Council Regulation EU 1099/2009 on the protection of animals at the time of killing.
The submitted study reports upon the use (mean + SD) of a current of 104.00 ± 3.88 mA, a voltage of 125.86 ± 3.28 V and a frequency of 589.78 ± 0.63 Hz using a square wave in alternating current (AC) with a 50 % duty cycle. These conditions were applied for 15 seconds to chickens under laboratory and slaughterhouse conditions.
The methodology and the data reported do not provide conclusive evidence that the combination of the proposed electrical frequency and current induced unconsciousness without exposing the chickens to avoidable pain and suffering, and some chickens did not remain unconscious for a sufficient time to prevent avoidable pain and suffering during slaughter.
EFSA stated in their report that it was doubtful that recovery of consciousness could be avoided prior to neck cutting and/or during bleeding. The minimum duration of unconsciousness was reported to be 11 seconds, which is too short to permit a feasible stun-to-stick interval. Further, it is also doubtful that recovery of consciousness could be avoided prior to neck cutting and/or during bleeding. The minimum time to resumption of breathing was reported to be 8 seconds following stunning.
Because the information provided in the study was incomplete and insufficient, it did not pass the eligibility phase of the assessment. The information provided was sufficient to conclude that the birds were not rendered immediately unconscious by the intervention. Application of a current less than that required inducing immediate unconsciousness causes pain, distress and suffering. The study failed to demonstrate absence of pain and suffering until onset of unconsciousness. The minimum duration of unconsciousness was too short to ensure unconsciousness until death by bleeding.
Toxicity studies in animals are conducted to identify any toxic effects of a substance prior to clinical use in humans. The document outlines various types of toxicity studies including acute, subacute, chronic, and lethality studies. Acute studies involve a single high dose to determine toxic effects over 14 days, while repeated dose studies like subacute and chronic studies administer multiple lower doses over weeks to years to identify target organ toxicity. Lethality studies determine the lethal dose for 50% of animals (LD50). Systemic toxicity parameters evaluated include effects on liver, kidney, heart and other organs. Toxicity studies provide safety information required for approval to conduct human clinical trials.
The document summarizes guidelines for skin sensitization toxicity studies using guinea pigs. It describes two common test types - adjuvant tests using Freund's Complete Adjuvant and non-adjuvant tests like the Buehler test. The preferred methods are the Guinea Pig Maximization Test and the non-adjuvant Buehler Test. Both tests involve induction exposure, a rest period, then challenge exposure to determine skin reaction responses. Precise protocols for each test are provided regarding animal selection, housing, dosing, observations, scoring, and reporting of results.
Introductiontoexperimentalpharmacologicalresearch 211008134020PHARMA IQ EDUCATION
This document provides an introduction to experimental pharmacological research. It discusses the definitions of pharmacology and experimental pharmacology. Experimental pharmacology involves both pre-clinical and clinical studies and can lead to new therapeutic agents or identification of toxic effects. The objectives of experimental pharmacology are to provide scientific knowledge of drugs, discover new drugs and techniques. Experimentation can be done in vivo using animal models like rats, mice and rabbits, or in vitro using isolated tissues, cells, or cell-free components. Selection of animal models considers factors like size, availability, sensitivity and species-specific responses. In vivo and in vitro methods are vital experimental tools in pharmacological research.
EU REACH regulation changed the way to do chemical risk assessment. All chemicals marketed or manufactured in the EU must have its own dossier. Non standard methods including alternatives to animal testing are accepted.
Half Italian, half English
Acute and repeated dose toxicity studies are important for determining the safety of pharmaceuticals intended for human use. Key details include:
- Acute toxicity studies involve single high doses to determine the median lethal dose (LD50) and maximum tolerated dose. Repeated dose studies last 14 days to 3 months.
- Studies are conducted in at least two species, often rats and non-rodents, to identify target organs and toxicity reversibility.
- Animals are observed for signs of toxicity and mortality. Necropsies and histopathology are performed to identify organ damage.
- Sub-acute studies last 14-90 days to determine effects of repeated administration and establish doses for longer term studies.
EFSA AHAW report on monitoring procedures at poultry slaughterhousesHarm Kiezebrink
The EFSA Panel on Animal Health and Welfare (AHAW) was asked to deliver scientific opinions on monitoring procedures at slaughterhouses for different animal species, stunning methods and slaughter without stunning. AHAW agreed that, although it is traditional to look for outcomes of unconsciousness in poultry following stunning, the risk of poor welfare can be detected better if bird welfare monitoring is focused on detecting consciousness, i.e. ineffective stunning or recovery of consciousness.
Therefore, the indicators were phrased neutrally (e.g. corneal reflex) and the outcomes were phrased either suggesting unconsciousness (e.g. absence of corneal reflex) or suggesting consciousness (e.g. presence of corneal reflex). This approach is commonly used in animal health studies (e.g. testing for the presence of a disease) but very new to animal welfare monitoring in slaughterhouses.
A toolbox of selected indicators is proposed to check for signs of consciousness in poultry after stunning with waterbaths or gas mixtures; a different toolbox of indicators is proposed for confirming death of the birds following slaughter without stunning.
This study investigates the possibility of a harmonized EU approach to assessing the added therapeutic value (ATV) of medicines. It finds that while ATV is not systematically assessed for marketing authorization, most EU countries do consider it as part of pricing and reimbursement decisions. The study reviews the EU legal framework, explores ATV practices across the EU-28, and provides an in-depth analysis of ATV in six countries. It closes by outlining recommendations for a possible European harmonization of ATV assessment within the current legal framework.
Endoscope surveillance - comparison of sampling techniquesOneLife SA
Objective : To compare different techniques of endoscope sampling to assess residual bacterial contamination.
Design : Diagnostic study
Setting : The endoscopy unit of a 1.100-bed university teaching hospital performing approximately 13 000 endoscopic procedures annually.
This document provides evidence-based recommendations for point-of-care lung ultrasound from an international panel of 28 experts. The panel reviewed literature from 1966 to 2011 using the GRADE method to determine evidence quality and develop recommendations. They discussed 73 proposed statements over three conferences using a modified Delphi technique and anonymous voting. Strong recommendations were made for 65 statements, and weak recommendations for 2 statements. The recommendations aim to standardize the application of lung ultrasound in clinical settings and provide a framework to guide its future use and research.
The European Medicines Agency has recommended approval of two H1N1 influenza vaccines, Focetria by Novartis and Pandemrix by GlaxoSmithKline. The agency expedited review of the vaccines to have them available for the upcoming flu season. Both vaccines were authorized using a "mock-up" approach where they were initially developed for a different flu strain but then updated for H1N1. The agency recommends a two dose schedule based on safety and efficacy data from over 6,000 subjects. Ongoing trials and post-approval monitoring will provide additional safety information.
This safety data sheet provides information on the product KASKARA EW. It contains a mixture of herbicides including 2,4-D EHE, dicamba DMA salt, and triclopyr butoxy ethyl ester. The product is classified as harmful if swallowed, may cause an allergic skin reaction, and is very toxic to aquatic life with long lasting effects. Personal protective equipment including gloves and eye protection should be worn when handling. Spills should be cleaned up promptly and the product stored away from heat, sparks and flames.
Washington Global Health Alliance Discovery Series
Supamit Chinsuttiwat
May 22, 2008
'Response to Avian Influenza and Preparedness for Pandemic Influenza: Thailand's Experience'
The document provides guidance for assessing the risk of tuberculosis (TB) transmission during air travel and conducting contact tracing of exposed passengers. It recommends contact tracing only after a careful risk assessment based on the infectiousness of the index patient, amount of exposure, and susceptibility of exposed individuals. Key criteria include the index case having confirmed infectious pulmonary TB, evidence of transmission to others, and exposed passengers sitting within two rows of the index case for a long-haul flight (≥8 hours). Contact tracing should follow WHO guidelines and investigate/manage exposed passengers for latent TB infection according to national guidelines. The guidance stresses avoiding unnecessary air travel for infectious TB patients and isolating/providing masks for suspected cases during flights.
This document provides guidelines for high quality control posts (CPs) for cattle, pigs, and sheep during long distance transport in Europe. It defines CPs as facilities approved by authorities where animals can rest, eat, drink and receive care during long journeys exceeding transport limits. The document outlines welfare benefits of CPs and potential disease risks if biosecurity is not maintained. It also summarizes relevant EU regulations regarding fitness for transport, biosecurity, and cleaning/disinfection of CPs between groups of animals.
This document provides guidelines for high quality control posts (CPs) for cattle, pigs, and sheep during long distance transport in Europe. It defines CPs as facilities approved by authorities where animals can rest, eat, drink and receive care during long journeys exceeding transport limits. The document outlines welfare benefits of CPs and potential disease risks if biosecurity is not maintained. It also summarizes relevant EU regulations regarding fitness for transport, biosecurity, and cleaning/disinfection of CPs between groups of animals.
This document provides safety information for the product Forester. It identifies Forester as a mixture containing the active ingredient Cypermethrin cis/trans at a concentration of 9.6-10.6%. Forester is classified as harmful if swallowed, may cause an allergic skin reaction, and is very toxic to aquatic life with long lasting effects. Emergency contact information is provided. Personal protective equipment is advised when handling Forester, which should be stored in a cool, well ventilated area away from heat and direct sunlight.
Overview on current practices of poultry slaughtering and poultry meat inspec...ABOHEMEED ALY
This report provides an overview of current practices for poultry slaughtering and meat inspection in the EU. It describes poultry slaughtering methods and the key steps in the process. It also discusses food chain information collection and its role in risk-based meat inspection. The report outlines specific laboratory testing conducted and conditions inspected for during ante-mortem and post-mortem examination. Finally, it provides country-specific details on how meat inspection is implemented and concludes with statistics on EU poultry meat production.
This document describes a study that developed a photoacoustic spectroscopy method to noninvasively monitor endogenous methane production in small laboratory animals and humans. The method was used to measure whole-body methane emission in mice and rats under normal conditions, after antibiotic treatment to reduce gut methanogens, and after lipopolysaccharide administration. Single-breath methane analyses were also performed on human participants. The study aimed to establish photoacoustic spectroscopy as a reliable tool for monitoring in vivo methane dynamics in response to various treatments.
1st Hepatitis E virus expert meeting at ECDC, IntroductionCornelia Adlhoch
The expert group meeting discussed hepatitis E virus (HEV) epidemiology, surveillance, and risks in the EU/EEA. HEV is an under-surveilled cause of hepatitis worldwide with different genotypes infecting people. While most EU cases were previously travel-related, HEV genotype 3 is now endemic in Europe with an animal reservoir. Surveillance varies between countries with no EU-wide system. The group aims to inventory HEV in Europe by surveying countries on surveillance methods and collecting case numbers to describe epidemiology and populations at risk. This will identify needs for EU guidance on diagnosis, risk assessment, and prevention strategies to respond to HEV health threats.
This report summarizes surveillance data on zoonoses (diseases transmitted between animals and humans), zoonotic agents, and foodborne outbreaks in the European Union in 2009. It finds that:
- Salmonellosis and campylobacteriosis were the most common zoonotic infections in humans. Salmonellosis cases decreased while campylobacteriosis cases increased slightly.
- Listeriosis cases in humans increased by 19.1% compared to 2008. Listeria was rarely found above safety limits in ready-to-eat foods.
- Over 5,500 foodborne outbreaks were reported causing nearly 49,000 illnesses, 4,356 hospitalizations and 46 deaths. The
CELEX 52014DC0188 EN TXT_Raport en.pdfssuser7f7ec8
This document provides a report on the application of the Regulation on advanced therapy medicinal products (ATMP Regulation) in the European Union from 2009 to 2013. It finds that while only four ATMPs have received marketing authorization so far, research activity is high with over 250 clinical trials and many requests for scientific advice. However, it also notes that many existing ATMPs continue to be used without marketing authorization under exemptions granted by Member States. The report concludes that more needs to be done to both ensure patient safety from unauthorized products and facilitate availability of new ATMPs to patients.
CELEX 52014DC0188 EN TXT_Raport en.pdfssuser7f7ec8
This document provides a report on advanced therapy medicinal products (ATMPs) in the European Union in accordance with Article 25 of Regulation (EC) No 1394/2007. It summarizes that while there is significant research on ATMPs, only a small fraction result in approved medicines. The regulation has established the Committee for Advanced Therapies and adapted regulatory requirements. To date, four ATMPs have received marketing authorization in the EU. The report analyses the impact of the regulation on research and development of advanced therapies.
This document provides a disease strategy for the management of an outbreak of sheep pox and goat pox in Australia. It summarizes the key information about the diseases, including that they are caused by viruses in the genus Capripoxvirus and primarily affect goats and sheep. Clinical signs include fever and lesions on the skin and in the mouth. The document outlines diagnostic criteria and treatment options, as well as principles of control and eradication that would be followed in an outbreak in Australia.
Similar to EFSA report on Low Atmoshere Pressure System to stun poultry (20)
Low Atmospheric Pressure Stunning is not a humane alternative to Carbon Dioxi...Harm Kiezebrink
I would like express gratitude to the HSA for their 20 years of tireless advocacy for improving pigs' welfare. Their efforts have empowered those seeking alternatives to carbon dioxide stunning. Over nearly 30 years, I've worked on animal welfare friendly stunning applications, particularly regarding stunning/slaughtering using nitrogen foam, and I believe I've found the definitive answer.
The industry originally adopted large-scale carbon dioxide stunning to optimize food production, reduce costs, and lower meat prices, which is only feasible with parallel processing (simultaneously stunning groups of pigs) rather than serial processing (stunning each pig individually). Electrocution is not viable for large-scale operations due to this need for parallel processing. Therefore, a replacement gas that lacks carbon dioxide's detrimental properties is needed, but only a few gases are suitable.
Additionally, the application of an alternative gas must adhere to several fundamental principles:
a) Applicability of the methods for stunning and killing pigs, including their scalability for large-scale application.
b) Description of the technical.
c) Animal welfare consequences associated with specific techniques, including welfare hazards (ABMs), animal-based indicators (ABIs), preventive and corrective measures, and the sufficiency of scientific literature in describing these consequences.
d) Applicability under field conditions.
Introducing a novel application for large-scale pig slaughter is complex and time-consuming before it can be expected, especially given the substantial economic and financial impact for the industry. However, there is hope on the horizon.
The alternative gas is nitrogen, and the application is based on using high-expansion foam filled with 100% nitrogen, applied in a closed container. Within a minute, all air is displaced by the foam, after which the container is sealed, and the foam is broken down with a powerful nitrogen pulse. This ensures that the foam does not affect the stunning process; the entire process can be visually and electronically monitored, and the residual oxygen level in the container is consistently below 2%. The container dimensions are identical to the gondolas used in the globally implemented carbon dioxide gondola system.
The integration of nitrogen foam technology into European regulation EU1099/2009 is nearing completion. All scientific and technical procedures have been submitted to the EU Commission, with finalization awaiting the presentation of EFSA's scientific opinion to the Commission and subsequent approval for inclusion. This final phase is anticipated to occur during the general meeting slated for June 2024.
This marks the first step toward replacing carbon dioxide in 25 years. Fingers crossed for the EU Commission's decision in June 2024!
Harm Kiezebrink
Independent Expert
Preventief ruimen bij vogelgriep in pluimveedichte gebieden en mogelijkheden ...Harm Kiezebrink
New Risk assessment model
The applications designed for farrow-to-weaner pig farms rely on a novel risk assessment model. This model, developed from a recent study, indicates that the likelihood of an undetected infection on nearby farms notably diminishes 7 to 14 days following the identification of the source farm.
This risk assessment model is based a Dutch study that is published by T.J. Hagenaars et al on June 30, 2023: “Preventief ruimen bij vogelgriep in pluimveedichte gebieden en mogelijkheden voor aanvullende bemonstering” (Preventive culling in areas densely populated with poultry, and possibilities for additional sampling).
According to this premise, instead of the standard depopulation approach of euthanizing pigs on-site, pigs beyond the immediate vicinity of infected farms are slaughtered.
Animal Health Canada is currently evaluating new strategies and technologies for managing large-scale emergency situations involving pigs. I have been actively involved in developing strategies and procedures aimed at implementing strict control measures for pig euthanasia during emergencies, with a focus on substantially reducing costs by avoiding unnecessary culling and destruction of healthy animals.
Opting for slaughtering over on-farm euthanasia not only reduces the operational burden on farms but also repurposes the pigs as a valuable protein source rather than considering them as animal waste. This approach assists in crisis management during widespread outbreaks, significantly reduces expenses, and simultaneously mitigates risks.
While this approach is influenced by the new EU regulations implemented since May 2022, it can be adapted for implementation within the context of any EU Member state, as well as in the USA and Canada.
Managing large-scale outbreaks at Farrow-to-Weaner FarmsHarm Kiezebrink
In the face of large-scale outbreaks of swine Influenza A Virus (swIAV), there's a call for exploring various strategies conducive to managing emergencies in field conditions.
Through subdivision, a customized approach can be embraced to enhance operational efficiency and effectiveness while mitigating the impact on individual farms. This tactic maximizes emergency deployment capacity and streamlines standard procedures. Moreover, leveraging the existing capacity of farming aids in alleviating scrutiny on animal welfare standards, presenting a notable advantage.
Nitrogen filled high expansion foam in open ContainersHarm Kiezebrink
On March 31, 2023 the US National Pork Board validated a study by Todd Williams, of Pipestone Veterinary Services, based on the use of high expansion nitrogen foam for the large-scale depopulation of all classes of swine, utilizing Livetec Systems Nitrogen Foam Delivery System (NFDS).
The high expansion foam produced by the Livetec Systems NFDS surrounds the animal in large bubbles filled with nitrogen with a base expansion ration of between 300 and 350 to 1, as mentioned on the information provided by the producer of the firefighting foam.
The Livetec technology, based on using Compressed Air Foam (CAF) filled with nitrogen instead of air for depopulating pigs, emerges within a critical landscape. The complexities of implementing effective emergency depopulation strategies for livestock, particularly swine, present multifaceted challenges. Livetec's approach relies on high expansion firefighting foam, aiming to euthanize pigs by submerging them in foam.
The Livetec system's claims about the effectiveness of nitrogen-filled high expansion foam for depopulating market pigs lack substantial evidence upon analysis. The discrepancy between the actual foam produced during field trials and the promised high expansion foam, coupled with the absence of concrete proof supporting the method's efficacy, discredits the technology's claims.
World bank evaluating the economic consequences of avian influenzaHarm Kiezebrink
Pandemics cause very serious loss of life, restrictions of freedom and serious economic damage. Potential pandemics all are related to our dealing with animals, both wild and domesticated.
In this Word Bank study of 2006, the effects of a severe HPAI pandemic (with a highly pathogenic avian influenza virus crossing the species barrier and infecting humans) predicted economic losses from 2-10% of the world economy.
The economic impact of the present COVID-19 crisis, caused by the SARS-CoV2 virus spreading from wild animals to humans, probably will reach the upper limits of this prediction even if the losses of life might be near the lower limits mentioned in the report (1,4 millions rather than 71 millions).
A common observation is that governments were late to react on the COVID-19 outbreak.
Pandemics are rare, so due to cost-benefit considerations emergency preparations do usually not get beyond an advisory (paperwork) phase. When an emergency eventually arises, the response is too late, too little, and with disastrous effects on animal and/or human welfare that could have been avoided. Relatively small, short-term financial savings result in big, long-term losses.
Protection against outbreaks cannot be achieved by political decisions during a crisis. Our dealing with animals, especially in animal production, must be inherently safe so that animal health and public health are protected.
This is recognized in the One Health strategy that has been adopted internationally.
An outbreak of animal disease occurs should be contained at a very early stage. This can only be realized if all farms have their own emergency plans, with equipment to deal with contagious diseases already present at the farm.
Anoxia: High expansion foam
The Anoxia method is unique for creating an environment without oxygen under atmospheric circumstances. High expansion foam is produced by mixing nitrogen and a mixture of water and specially developed high expansion detergent, with an expansion rate upto 1:1000, meaning that 1 litre of water/foam agent mix expands up to 1 m3 foam. Due to the specially designed foam generator, the high expansion foam bubbles are filled with a > 99% concentration of nitrogen. The oxygen level surrounding the animal drops from 21% in atmospheric air to < 1 % once the animal is submerged in the foam.
Anoxia: convulsions, but no stress or pain
The animals need a constant supply of oxygen to the brain. Applying Anoxia foam, the oxygen is replaced by nitrogen. As a result the nitrogen level is raised to > 99% and the oxygen level is lowered to < 1%. Considering the natural reaction to sudden lack of oxygen the animal is rendering quickly into unconsciousness. As a consequence, behavioral indicators like loss of posture and convulsions will appear. With this in mind, unconscious animals are insensitive to perceive unpleasant sensations like pain.
Anoxia: How Anoxia foam is created
A mixture of 97% water and 3% high expansion foam agent is sprayed into the Anoxia foam generator, creating a thin film on the outlet of the generator. At the same time, nitrogen is added with overpressure into the foam generator. The nitrogen expands when it exits the generator, creating robust high expansion foam. The high expansion foam bubbles are filled with > 99% nitrogen.
Anoxia: Single foam generator systems
In practice, one Anoxia foam generator creates a volume of up to 750 liter of high expansion foam per minute. This volume is more than sufficient to fill a wheelie-bin container within 30 seconds. The most common container volumes are: M size - 240 liter; L size - 340 liter; and XL size - 370 liter. The choice of the volume of the container depends of the size of the animal and/or the number of animals that need to be stunned/killed. A lid with a chiffon that seals the container. As soon as the foam exits the chiffon, the gas supply is stopped and the chiffon is closed. The nitrogen gas concentration in the container remains at 99%.
Although commonly used in other settings, defining animal welfare as part of a corporate CSR setting is not new.
There are many ways to define CSR. What they have in common is that CSR describes how companies manage their business processes to produce an overall positive impact on society. The phenomenon CSR is a value concept that is susceptible to particular ideological and emotional interpretations. Different organizations have framed different definitions - although there is considerable common ground between them.
Some important national players of the food chain at different steps (mainly food retailers and food services) have included animal welfare in their CSR.
The Anoxia technique is developed as alternative for existing animal stunning methods that are based on the use of CO2, electrocution, neck dislocation, captive-bolt, as well as killing methods like de-bleeding and maceration.
In the past 10 years, Wageningen University and University of Glasgow conducted several studies that proved that the technique could be applied successfully for culling poultry (Proof of Principle Anoxia Technique). This was the start of the development of several applications based on the Anoxia principle, using high expansion foam filled with >99% Nitrogen that are now introduced for:
1. Stunning and killing of sick and cripple killing piglets less than 5 kg
2. Stunning and killing of sick or cripple poultry (especially poultry > 3kg) who need to be killed on the farm by the staff for welfare purposes (avoiding unnecessary stress or pain)
3. Stunning and killing poultry that arrives on the slaughterhouse but that are unfit to be slaughtered (due to injuries occurred during transportation – providing signs of possible illness etc.)
4. Stunning and killing of male pullets at the hatchery
5. Stunning and killing of half-hatched chickens and embryos in partly-hatched eggs, before destruction
6. Stunning and killing parent stock poultry
7. Killing of animals that has been stunned (captive bolt – blow-on-the-head method, etc.) replacing killing by de-bleeding
8. Culling of ex-layers
9. Culling of poultry for disease control purposes
Last November we started the launch of the commercialization of the Anoxia applications in Holland, Germany and Sweden, focusing on the areas where a solution is most needed: piglets (< 5kg) and poultry (> 3kg) on farms.
Since November 2016, the introduction of these applications took place in Holland, Germany, Sweden and Denmark
World Health Organization director- general Margaret Chan Fung Fu-chun warns bird flu H7N9 is particularly worrying as it could be a flu pandemic strain. This is because H7N9 is unique as it does not make chickens sick but is deadly in humans. Sick birds could usually provide early warning for imminent outbreaks, Chan told The Standard. This comes as Macau reported its first human case of H7N9 yesterday. "The biggest challenge for the world is the next influenza pandemic," Chan said.
Laves presentation practical experiences in the culling of poultry in germanyHarm Kiezebrink
This presentation, based on the practical experiences in culling poultry in Germany, gives an overview of the culling techniques currently in use in Germany. It is presented by dr. Ursula Gerdes, dr. Josef Diekmann and ing. Rainer Thomes.
LAVES is the Lower Saxony State Office for Consumer Protection and Food Safety, located in Oldenburg, Germany. With around 900 employees they are entrusted with tasks in the areas of food and utensil inspection, feed inspection, meat hygiene, veterinary drug monitoring, eradication of animal diseases, disposal of animal by-products, animal welfare, ecological farming, market surveillance and technical process monitoring.
Berg et al. 2014 killing of spent laying hens using co2 in poultry barnsHarm Kiezebrink
September 2015: In Sweden, spent laying hens are killed either by traditional slaughter; on-farm with CO2 in a mobile container combined with a grinder; or with CO2 stable gassing inside the barn. The number of hens killed using the latter method has increased. During these killings a veterinarian is required to be present and report to the Swedish Board of Agriculture.
Data were registered during four commercial killings and extracted from all official veterinary reports at CO2 whole-house killings in 2008–2010. On-farm monitoring showed that temperature decreased greatly and with high variability. The time until birds became unconscious after coming into contact with the gas, based on time until loss of balance, was 3–5 min.
Veterinary reports show that 1.5 million laying hens were killed, in 150 separate instances. The most common non-compliance with legislation was failure to notify the regional animal welfare authorities prior to the killings. Six out of 150 killings were defined as animal welfare failures, eg delivery of insufficient CO2 or failure to seal buildings to achieve adequate gas concentration.
Eleven were either potentially or completely unacceptable from the perspective of animal welfare. We conclude that, on the whole, the CO2 whole-house gas killing of spent hens was carried out in accordance with the appropriate legislation. Death was achieved reliably.
However, there remain several risks to animal welfare and increased knowledge would appear vital in order to limit mistakes related to miscalculations of house volume, improper sealing or premature ventilation turn-off.
The latest outbreak of High Pathogen Avian Influenza in the USA and Canada in the spring of this year and the inability to avoid animal welfare catastrophes ultimately proves that new emergency response strategies are needed. Strategies that are based on taking away the source of infection instead of killing as many animals as possible within 24 hours, regardless the consequences.
The statement that “It’s possible that human infections with these viruses may occur” and that “these viruses have not spread easily to other people” is confusing. Humans can become infected without showing clinical signs. They can become the major carrier of the infection.
Especially during depopulation activities, viruses easily transmit through responders. Tasks like taking layers out of their cages and transport the birds manually through the narrow walkways between the cages, and disposal of infected animals are specific risks that need to be avoided. Simply switching of the electricity so that sick birds don’t have to be handled is not the solution.
Although humans are supposed to be less susceptible, they can become carrier of the virus. Only the highest level of biosecurity could prevent the transmission through the humans and materials that have been in direct contact with infected animals and materials.
Simply switching of the electricity so that sick birds don’t have to be handled is not the solution. Avoid killing animals is always the better option and in Germany, the discussion on the strategy based on neutralizing risks and is in the making. Avoiding situations demands a proactive role of the poultry industry.
Ventilation Shutdown: who takes the responsibility to flip the switch?Harm Kiezebrink
On September 18, 2015 the USA Government and the American egg producers announced that they would accept the Ventilation shutdown method as a method of mass destruction of poultry when other options, notably water-based foam and CO2, are not available for culling at the farm within 24-36 hours. This is actually the case on all caged layer farms in the USA, in particular in Iowa.
The Ventilation shutdown method consists of stopping ventilation, cutting off drinking water supply, and turning on heaters to raise the temperature in the poultry house to a level between 38 Celsius and 50 Celsius. Birds die of heat stress and by lack of oxygen in a process that easily takes over after a period of at least 3 days. Ventilation shutdown is a killing method without prior stunning of the birds, and as such is contrary to all international Animal Welfare standards.
Animal welfare specialists in disease control strongly oppose this introduction of the cruelest method of killing poultry that lost their economic value. The Humane Society (HSUS) described it as the “inhumane mass baking of live chickens”. With adequate preparation the alternative methods, like the water-based Anoxia foam method, can be available at each farm for immediate use in case of an outbreak. The ban of the Ventilation shutdown method should therefore be maintained and the Anoxia method should be further developed so that is suitable for application to caged layers and turkeys. In Germany, such a system is currently under development and will become commercially available soon.
The poultry industry in the USA ignores this development and asks for a formal approval of the Ventilation Shutdown method. Speaking on August 19, 2015, during the United Egg Producers (UEP) national briefing webinar, UEP President Chad Gregory explained that much research is being done concerning the feasibility of such a depopulation program.
“The government, the producers, the states and UEP, we all recognize that depopulation is going to have to happen faster and ideally within 24 hours.”
Quick depopulation of affected flocks is important, Gregory said, because the sooner a flock is depopulated, the risk of the virus going into fans and out into the atmosphere becomes smaller. Gregory said ventilation shutdown – if approved – would probably only be used in a worst-case scenario or when all other euthanasia options have been exhausted. Gregory did not elaborate on how to adequately prevent outbreaks and how to promote more animal-friendly methods.
In order to become one step ahead of an outbreak of high pathogen diseases like the current H5N2, the veterinary authorities need to stop the outbreak immediately after the first signals occur. Strict and thorough biosecurity measures are the most fundamental feature to protect poultry flocks on farms.
Without functional culling techniques, the options to effectively and efficiently cull in average more than 925,000 chickens per farm (in Iowa, USA) are limited: either by macerating the chickens alive – or by ventilation shut-down (closing down all ventilation, placing heaters inside the house, and heat the entire house to a temperature higher than 600 C).
Although both methods cause death of the birds, it has not been proven to be effective nor efficient. The primary goal to slowdown outbreaks and bring it to a complete stop but macerating live birds and killing them by heat stress and lack of oxygen would be against all International Animal Welfare standards.
Animal welfare specialists in disease control strongly oppose against the introduction of these most cruel methods of killing poultry and argue that the ban on these methods should be maintained and alternative methods need to be considered.
FLI Seminar on different response strategies: Stamping out or NeutralizationHarm Kiezebrink
During this spring, American poultry producers are losing birds by the millions, due to the High Pathogenic Avian Influenza outbreaks on factory farms. USDA APHIS applied the stamping out strategy in an attempt to prevent the flu from spreading.
With stamping out as the highest priority of the response strategy, large numbers of responders are involved. With in average almost 1 million caged layers per farm in Iowa, there is hardly any room for a proper bio security training for these responders. And existing culling techniques had insufficient capacity, the authorities had to decide to apply drastic techniques like macerating live birds in order to take away the source of virus reproduction.
This strategy didn't work; on the contrary. Instead of slowing down the spreading of the virus, the outbreaks continue to reoccur and have caused death and destruction in 15 USA states, killing almost 50 million birds on mote than 220infected commercial poultry farms, all within a very small time frame.
The question is whether the priority of the response strategy should be on neutralizing the transmission routes instead of on stamping out infections after they occur. All indicators currently point out into the direction that the industry should prioritize on environmental drivers: the connection between outbreaks and wild ducks; wind-mediated transmission; pre-contact probability; on-farm bio security; transmission via rodents etc.
Once the contribution of each transmission route has been determined, a revolutionary new response strategy can be developed based on the principle of neutralizing transmission routes. Neutralizing risks means that fully new techniques need to be developed, based on culling the animals without human – to – animal contact; integrating detergent application into the culling operations; combining culling & disposal into one activity.
This new response strategy will be the main subject of the FLI Animal Welfare and Disease Control Seminar, organized at September 23, 2015 in Celle, Germany
Dossier transmission: Transmission of Avian Influenza Virus to DogsHarm Kiezebrink
This document reports on the transmission of an avian influenza virus (H3N2) to dogs in South Korea. Several dogs exhibited severe respiratory disease and three genetically similar canine influenza virus strains were isolated. Experimental infection of beagles demonstrated that the virus could be transmitted between dogs and cause clinical signs like fever and lung lesions. The canine respiratory tract was found to contain receptors for binding avian influenza viruses, suggesting potential for direct transmission from poultry. This provides evidence that dogs may play a role in interspecies transmission of influenza viruses.
Spatio temporal dynamics of global H5N1 outbreaks match bird migration patternsHarm Kiezebrink
This document analyzes the spatiotemporal patterns of H5N1 avian influenza outbreaks globally between 2003 and 2006. It identifies three phases of the H5N1 epidemic and uses space-time cluster analysis to detect six disease cluster patterns along major bird migration flyways. The matching of outbreak clusters with wild bird migration patterns suggests wild birds may play an important role in long-distance spread of H5N1. Short-distance spread is also potentially linked to wild birds spreading the virus at sites where they overwinter or migrate through.
Spatial, temporal and genetic dynamics of H5N1 in chinaHarm Kiezebrink
The spatial spread of H5N1 avian influenza, significant ongoing mutations, and long-term persistence of the virus in some geographic regions has had an enormous impact on the poultry industry and presents a serious threat to human health.
This study revealed two different transmission modes of H5N1 viruses in China, and indicated a significant role of poultry in virus dissemination. Furthermore, selective pressure posed by vaccination was found in virus evolution in the country.
Phylogenetic analysis, geospatial techniques, and time series models were applied to investigate the spatiotemporal pattern of H5N1 outbreaks in China and the effect of vaccination on virus evolution.
Results showed obvious spatial and temporal clusters of H5N1 outbreaks on different scales, which may have been associated with poultry and wild-bird transmission modes of H5N1 viruses. Lead–lag relationships were found among poultry and wild-bird outbreaks and human cases. Human cases were preceded by poultry outbreaks, and wild-bird outbreaks were led by human cases.
Each clade has gained its own unique spatiotemporal and genetic dominance. Genetic diversity of the H5N1 virus decreased significantly between 1996 and 2011; presumably under strong selective pressure of vaccination. Mean evolutionary rates of H5N1 virus increased after vaccination was adopted in China.
Different environmental drivers of H5N1 outbreaks in poultry and wild birdsHarm Kiezebrink
Different environmental drivers operate on HPAI H5N1 outbreaks in poultry and wild birds in Europe. The probability of HPAI H5N1 outbreaks in poultry increases in areas with a higher human population density and a shorter distance to lakes or wetlands.
This reflects areas where the location of farms or trade areas and habitats for wild birds overlap. In wild birds, HPAI H5N1 outbreaks mostly occurred in areas with increased NDVI and lower elevations, which are typically areas where food and shelter for wild birds are available. The association with migratory flyways has also been found in the intra-continental spread of the low pathogenic avian influenza virus in North American wild birds. These different environmental drivers suggest that different spread mechanisms operate.
Disease might spread to poultry via both poultry and wild birds, through direct (via other birds) or indirect (e.g. via contaminated environment) infection. Outbreaks in wild birds are mainly caused by transmission via wild birds alone, through sharing foraging areas or shelters. These findings are in contrast with a previous study, which did not find environmental differences between disease outbreaks in poultry and wild birds in Europe.
H5N8 virus dutch outbreak (2014) linked to sequences of strains from asiaHarm Kiezebrink
Genetic analysis of influenza A(H5N8) virus from the Netherlands indicates that the virus probably was spread by migratory wild birds from Asia, possibly through overlapping flyways and common breeding sites in Siberia. In addition to the outbreak in the Netherlands, several other outbreaks of HPAI (H5N8) virus infections were reported in Europe at the end of 2014 after exponentially increasing deaths occurred in chicken and turkey flocks.
Genetic sequences submitted to the EpiFlu database indicated that the viruses from Europe showed a strong similarity to viruses isolated earlier in 2014 in South Korea, China, and Japan. An H5N8 virus isolated from a wigeon in Russia in September 2014 is located in the phylogenetic tree near the node of all sequences for H5N8 viruses from Europe.
In regard to time, this location fits the hypothesized route of H5N8 virus introduction into Europe. Furthermore, for several reasons, it is highly likely that the introduction of HPAI (H5N8) virus into the indoor-layer farm in the Netherlands occurred via indirect contact.
First, despite intensive monitoring, H5N8 viruses have never been detected in commercial poultry or wild birds in the Netherlands.
Second, when the virus was detected, the Netherlands had no direct trade contact with other European countries or Asia that might explain a route of introduction.
Third, because of the severity of disease in galliforms, outbreaks of H5N8 in the Netherlands before November 2014 would have been noticed.
An astonishing, first-of-its-kind, report by the NYT assessing damage in Ukraine. Even if the war ends tomorrow, in many places there will be nothing to go back to.
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2. Low atmosphere pressure system for stunning poultry
EFSA Journal 2014;12(1):3488 2
SUMMARY
Following a request from the European Commission, the EFSA Panel on Animal Health and Welfare
(AHAW) was asked to deliver a scientific opinion on the use of a low atmosphere pressure system
(LAPS) for stunning poultry prior to slaughter. Four documents were provided by the EC as the basis
for an assessment of the extent to which the LAPS is able to provide a level of animal welfare at least
equivalent to that ensured by the current allowed methods for stunning poultry. In the case that the
outcome of the assessment was positive, the EC requested an assessment of the conditions under
which the LAPS could be used in a commercial context.
The LAPS is described as rendering poultry unconscious by gradually reducing oxygen tension in the
atmosphere leading to progressive hypoxia in the birds. This intervention is not permitted in the EU. In
order to be allowed in the EU, new stunning methods must ensure a level of welfare at least equivalent
to that of the methods already provided in Council Regulation 1099/2009.
In this opinion, the term „acceptable alternative‟ is defined as an alternative stunning intervention that
is at least as good as those listed in the Council Regulation (EC) 1099/2009. In particular, for
interventions that do not induce immediate unconsciousness, the alternative procedure should ensure
1) absence of pain, distress and suffering until the onset of unconsciousness and, 2) that the animal
remains unconscious and insensible until death.
Following the adoption of two Scientific Opinions on the stunning of rabbits by CO2 and the electrical
stunning of lambs (EFSA, 2013a, b), the EFSA Panel on Animal Health and Welfare developed a
guidance on the process and criteria applied by EFSA to assess studies evaluating the effectiveness of
stunning interventions regarding animal protection at the time of killing (EFSA, 2013c). The approach
and criteria defined in that guidance were applied here.
The eligibility criteria that must be fulfilled by submitted studies are set in the EFSA guidance (EFSA,
2013c) and focus on the intervention and the animal welfare outcome. For the LAPS intervention, the
key parameters about which information must be provided are: animal density, duration of
intervention, rate of decompression, rate of changes in partial pressure of oxygen,
temperature/humidity/illumination of the chamber, maximum stun-to-stick/kill interval(s) and
calibration of the LAPS equipment and monitoring system. For the outcome of the LAPS intervention,
a description of the onset and duration (until death) of unconsciousness and insensibility and a
demonstration of the absence of pain, distress and suffering until the loss of consciousness and
sensibility must be provided. As requested by the Commission, the assessment focussed on the animal
welfare aspects of the LAPS. None of the four studies met the eligibility criteria and, therefore, neither
reporting nor methodological quality were assessed. It is unclear from the submitted studies whether
the rate of decompression used in the LAPS induces unconsciousness and death without causing
avoidable pain and suffering in poultry. Since the rate of decompression was not fully described in the
submitted studies, it was not possible to evaluate consistency with other sources of information.
3. Low atmosphere pressure system for stunning poultry
EFSA Journal 2014;12(1):3488 3
TABLE OF CONTENTS
Abstract .................................................................................................................................................... 1
Summary .................................................................................................................................................. 2
Table of contents ...................................................................................................................................... 3
Background as provided by the European Commission........................................................................... 4
Terms of reference as provided by the European Commission................................................................ 4
Assessment............................................................................................................................................... 5
1. Introduction ..................................................................................................................................... 5
2. Approach ......................................................................................................................................... 6
3. Eligibility criteria............................................................................................................................. 9
3.1. Specification of eligibility criteria .......................................................................................... 9
3.1.1. Intervention......................................................................................................................... 9
3.1.2. Outcome ........................................................................................................................... 10
3.2. Assessment of the eligibility criteria of the submitted studies.............................................. 13
3.2.1. Intervention....................................................................................................................... 13
3.2.2. Outcome ........................................................................................................................... 14
4. Reporting quality........................................................................................................................... 15
4.1. Assessment of the reporting quality of the submitted studies based on the selected
parameters.......................................................................................................................................... 15
5. Methodological quality.................................................................................................................. 15
5.1. Quality assessment of the internal validity of the submitted studies .................................... 16
5.2. The extent to which the findings are consistent with other sources of information;............. 16
Conclusions and recommendations........................................................................................................ 16
Conclusions............................................................................................................................................ 16
Recommendations .................................................................................................................................. 16
References.............................................................................................................................................. 16
Appendix. Assessment of the eligibility criteria .................................................................................... 19
Glossary and abbreviations .................................................................................................................... 27
4. Low atmosphere pressure system for stunning poultry
EFSA Journal 2014;12(1):3488 4
BACKGROUND AS PROVIDED BY THE EUROPEAN COMMISSION
Article 4 (2) of Council Regulation (EC) No 1099/2009 on the protection of animals at the time of
killing4
allows the Commission to amend Annex I to this Regulation as to take into account scientific
and technical progress on the basis of an opinion of the EFSA. Any such amendments shall ensure a
level of animal welfare at least equivalent to that ensured by the existing methods.
At present, the use of low atmosphere pressure system is not allowed for stunning poultry.
The Commission has received a request from a private business operator to allow the use of low
atmosphere pressure system as a method for stunning poultry. This request is supported by four
scientific publications (see attachment).
In order to reply to this request, the Commission would like to request the EFSA to review the
scientific publications provided and possibly other sources if available and assess to which extent the
system proposed for stunning poultry is able to provide a level of animal welfare at least equivalent to
that ensured by the current allowed methods and, in case of favourable reply, under which conditions.
TERMS OF REFERENCE AS PROVIDED BY THE EUROPEAN COMMISSION
The Commission therefore considers it opportune to request the EFSA to give an independent view on
the use of low atmosphere pressure system for stunning poultry.
The scope of this request is limited to the stunning of broiler chicken for slaughter (i.e. killing for
human consumption).
The EFSA will give its view on the four scientific publications attached to this request with a focus
on the following issues:
− The extent to which the use of low atmosphere pressure system is, in principle, an
acceptable alternative for the stunning of broiler chicken compared to the welfare
advantages/disadvantages related to other stunning methods allowed in the EU for broiler
chicken;
− The extent to which the findings are consistent with other sources of information;
− Requirements attached to the use of low atmosphere pressure system;
− The extent to which the findings may be valid under commercial conditions in the EU.
4
COUNCIL REGULATION (EC) No 1099/2009 of 24 September 2009 on the protection of animals at the time of killing.
OJ L 303, 18.11.2009, p. 1-30.
5. Low atmosphere pressure system for stunning poultry
EFSA Journal 2014;12(1):3488 5
ASSESSMENT
1. Introduction
The low atmospheric pressure system (LAPS) is a new intervention that has been claimed to render
poultry unconscious by gradually reducing oxygen tension in the atmosphere leading to progressive
hypoxia in animals. This intervention is not permitted in the EU. In order to be allowed in the EU, new
stunning methods must ensure a level of welfare at least equivalent to that ensured by the methods
already provided in Council Regulation 1099/2009.
Following the adoption of two Scientific Opinions on the stunning of rabbits by CO2 and the electrical
stunning of lambs (EFSA, 2013a, b), the EFSA Panel on Animal Health and Welfare (AHAW Panel)
developed a guidance on the process and criteria applied by EFSA to assess studies evaluating the
effectiveness of stunning interventions regarding animal protection at the time of killing (EFSA,
2013c). On the receipt of this mandate, its terms of reference were discussed with the European
Commission services and the following clarifications were made.
EFSA will give its independent view on the findings of the four studies submitted to the Commission:
STUDY 1 - “Physiological responses to low atmospheric pressure stunning (LAPS) and
implications for welfare” - McKeegan et al., in press5
, from now referred as “study 1”;
STUDY 2 - “ A new humane method of stunning broilers using low atmospheric pressure” -
Vizzier-Thaxton et al. (2010), from now on referred to as “study 2“;
STUDY 3 - “The effects of low atmosphere stunning and deboning time on broiler breast meat
quality” - Schilling et al. (2012), from now on referred to as “study 3“;
STUDY 4 - “The Effects of Low-Atmosphere Stunning and Deboning Time on Broiler Breast
Meat Quality” - Battula et al. (2008), from now on referred to as “study 4“.
The assessment of the submitted studies was carried out in a manner analogous to the approach
followed in previously adopted opinions and as outline in the EFSA AHAW Panel guidance (EFSA,
2013c). The assessment focuses on:
TOR 1: the extent to which the use of low atmosphere pressure system is, in principle, an
acceptable alternative for the stunning of broiler chicken compared to the welfare
advantages/disadvantages related to other stunning methods allowed in the EU for broiler
chicken;
TOR 2: the extent to which the findings are consistent with other sources of information.
An assessment of the evidence of validity under commercial conditions as well as the definitions of
requirements attached to the use of LAPS would be performed only if the outcome of the assessment
of suitability was positive.
The term “acceptable alternative” is defined as an alternative stunning intervention that is at least as
good as those listed in the Council Regulation (EC) 1099/2009. In particular, for interventions that do
not induce immediate unconsciousness, the alternative procedure should ensure 1) absence of pain,
distress and suffering until the onset of unconsciousness, and 2) that the animal remains unconscious
and insensible until death.
5
Published later as McKeegan et al, 2013.
6. Low atmosphere pressure system for stunning poultry
EFSA Journal 2014;12(1):3488 6
2. Approach
The submitted studies were assessed following the approach and specific criteria outlined in the
guidance document (EFSA, 2013c), summarized below. The assessment was first conducted
independently by each working group member. The individual assessments were then discussed to
reach a consensus on parameters for which the experts had expressed different opinions.
As requested by the Commission, the assessment focussed on the animal welfare aspects of the LAPS.
Further, the assessment only refers to the stunning procedure itself; it does not take into account pre-
stunning phases. The outcome of the assessment applies only to whether the assessed study is eligible
to be passed on to the next phase in the process: a full assessment of the animal welfare implications
of the proposed alternative stunning intervention, including both pre-stunning and stunning phases,
and an evaluation of the quality, strength and external validity of the evidence presented (Figure 1).
Figure 1: The approach of the assessment of studies evaluating alternative stunning methods
(X=exclusion of study from further assessment; in this case a description of the shortcomings and
indications where improvements are required before the study can be assessed further, will be
provided)
Where a study meets the necessary eligibility criteria regarding the description of the intervention and
outcome, then a reporting quality assessment is carried out.
Research studies evaluating stunning methods require well controlled studies under laboratory
conditions (Figure 2, I) as a first step, to characterise the animals‟ responses (unconsciousness,
absence of pain) using the most sensitive and specific methods available (e.g. EEG, blood samples)
and to establish the correlations between these measurements and non-invasive parameters that can be
applied in slaughterhouses (Figure 2). The second step, studies under slaughterhouse conditions
(Figure 2, II) is intended to assess whether the results obtained in the laboratory can also be achieved
in a slaughterhouse context. The eligibility criteria will be applied to both steps of the research on
stunning methods. Information obtained on other species can be used as an indication, but should be
confirmed in the species under investigation because coping strategies, pain thresholds and tolerances
are species and individual specific.
7. Low atmosphere pressure system for stunning poultry
EFSA Journal 2014;12(1):3488 7
Figure 2: Recommended approach for research on stunning methods
It is important to note that in controlled environment studies electroencephalograms (EEGs) or
electrocorticograms (ECoGs) should be used to demonstrate the effectiveness of a given stunning
intervention (Figure 2, B). Indicators for recognising a successful stun should be applied in
slaughterhouse settings, after their correlation with EEGs has been demonstrated in controlled
environment studies (Figure 2, G). Further details can be found in the guidance document (EFSA.
2013c).
Studies on stunning methods should explain in detail how and when the onset of unconsciousness and
insensibility is measured (Figure 2, B, C, G, H). In the case of EEGs (or ECoGs), all parameters
crucial to the assessment of the electroencephalogram data should be specified (e.g. the EEG recording
electrode position on the skull or on the brain itself, the configuration of the electrode
(transhemispheric or from the same hemisphere of the brain), the background noise filtration method
employed in the data acquisition and analysis). In order to estimate quantitative changes occurring in
the EEG (or ECoGs), the method used to derive the transformations of electrical brain signals must be
described (Figure 2, B).
Moreover, it should be explained how and when the animal-based measures were recorded and
analysed (Figure 2, G, H, I). Furthermore, data should be provided at the individual animal level.
For any intervention that does not lead to an immediate onset of unconsciousness and insensibility, the
time to loss of consciousness after the application of the stunning intervention and signs of pain,
distress and suffering until the onset of unconsciousness should be recorded in all animals and
reported as individual animal level data or mean or median and range and standard deviation or
interquartile range (Figure 2, B, C, G, H).
8. Low atmosphere pressure system for stunning poultry
EFSA Journal 2014;12(1):3488 8
As described in detail in the EFSA guidance and as presented in Figure 1, the following steps are to be
followed:
1) Eligibility criteria
Council Regulation (EC) No 1099/2009 defines “stunning” in Article 2(f) as “any intentionally
induced process which causes loss of consciousness and sensibility without pain, including any
process resulting in instantaneous death”. Furthermore, Article 4 on stunning methods regulates that
“animals shall only be killed after stunning in accordance with the methods and specific requirements
related to the application of those methods set out in Annex I of the Regulation” and “that the loss of
consciousness and sensibility shall be maintained until the death of the animal”. The methods referred
to in Annex I that do not result in instantaneous death shall be followed as quickly as possible by a
procedure ensuring death such as bleeding, pithing, electrocution or prolonged exposure to anoxia.
Most of the methods listed in Annex 1 cause immediate onset of unconsciousness, with the exception
of controlled atmosphere- or gas-stunning methods.
The eligibility criteria that must be fulfilled by submitted studies related to LAPS were set in the
EFSA guidance (EFSA, 2013c) and focus on the intervention and the outcome:
a) For the intervention:
This intervention is not currently approved for use in the EU, and therefore, no parameters are defined
by Council Regulation (EC) No 1099/2009. The parameters and components were defined by the
experts on stunning methods consulted during the preparation of this opinion.
b) For the outcome:
A. Onset of unconsciousness and insensibility OR
B. Absence of pain, distress and suffering until the loss of consciousness and sensibility
AND
C. Duration of the unconsciousness and insensibility (until death).
To allow assessment of new or modified legal stunning methods, the minimum criteria that fully
define and characterise the stunning intervention were defined using previously published scientific
data. Regarding measures of the outcome, the onset and duration of unconsciousness and insensibility
should be recorded and reported. If the onset of unconsciousness and insensibility achieved by the
stunning intervention is not immediate, then the absence of pain, distress and suffering until the loss of
consciousness and sensibility must also be recorded and reported.
2) Reporting quality criteria
Reporting quality will only be assessed when the scientific study has passed the eligibility assessment
(Figure 1). Inconsistencies in the reporting of scientific studies – which make it difficult to assess and
compare them - have been identified in human and veterinary medicine. Therefore, the guidance
document identified the relevant parameters that will be used as the basis for assessing the reporting
quality of submitted studies on stunning methods.
3) Methodological quality criteria
The methodological quality of the submitted study will be assessed only if the eligibility and reporting
quality criteria are met (Figure 1). In that case, the information provided in the study will be used to
identify and assess possible biases (e.g. selection, attrition and performance bias) that might affect the
study‟s internal validity.
9. Low atmosphere pressure system for stunning poultry
EFSA Journal 2014;12(1):3488 9
4) Possible outcomes of the assessment
As described in the guidance document, two outcomes of the assessment are possible:
a) The criteria regarding eligibility, reporting and methodological quality are fulfilled.
This means that the study on the new or modified legal intervention provides sufficient detail
regarding the intervention and the outcome to allow for a conclusion to be reached about the suitability
(or lack thereof) of the intervention. In that case, a full assessment of the animal welfare implications
of the proposed alternative stunning intervention, including both pre-stunning and stunning phases,
and an evaluation of the quality, strength and external validity of the evidence presented would be
carried out at the next level of the assessment (Figure 1).
b) The criteria regarding eligibility, reporting and methodological quality are not fulfilled.
This means that the study does not provide sufficient detail regarding the intervention and the outcome
to allow for a conclusion to be reached about its suitability (or lack thereof). In that case, the
assessment report would highlight the shortcomings and indicate where improvements are required
before the study could be assessed further.
3. Eligibility criteria
3.1. Specification of eligibility criteria
3.1.1. Intervention
LAPS is a stunning system where animals are rendered unconscious prior to slaughter by gradually
reducing oxygen tension in the atmosphere to achieve a progressive hypoxia. The induction of
unconsciousness with LAPS is not instantaneous. This intervention is not currently approved for use in
the EU, and therefore, no parameters are defined by Council Regulation (EC) No 1099/2009. The
parameters and components listed in Table 1 were defined by the experts on stunning methods
consulted during the preparation of this opinion.
Table 1: Suggested parameters to be provided when applying an intervention based on low
atmosphere pressure system for stunning poultry as determined by the EFSA ad-hoc expert
working group (EFSA, 2013c)
Parameter Component Description
(all specifications should be in internationally recognised units)
Animal density Animal species/ age/ type
and stocking density
(number/m2
and kg of
body weight/ m2
)
Specify the animal density in the crate or containers
during the decompression.
Duration of
interventiona
Time to achieve the
target pressures and
corresponding partial
pressure of oxygen in a
single-phase system or
multi-phase systemb
Report the time elapsing until animals are exposed to
the targeted pressure and corresponding partial
pressure of oxygen.
Report the duration of exposure to the target pressure
and corresponding partial pressure of oxygen;
If animals are exposed to a multi-stage system,
report the target pressure in each stage and the
duration of the exposure to each step as well as the
transition time between each step.
Rate of decompression Time/pressure treatment
graphic representation
Describe the rate at which pressure changes are
achieved in the chamber through a time/pressure
curve.
If decompression is achieved in more than one step,
the profile for each step should be described.
Re-pressurisation of the chamber prior to opening of
door should be described and any incidence of birds
10. Low atmosphere pressure system for stunning poultry
EFSA Journal 2014;12(1):3488 10
Parameter Component Description
(all specifications should be in internationally recognised units)
surviving the treatment should be reported.
Rate of changes in
partial pressure of
oxygen
Time/partial pressure of
oxygen treatment graphic
representation
Describe the rate at which partial pressure of oxygen
changes in the chamber in relation to the rate of
decompression.
If decompression is achieved in more than one step,
the profile for each step should be described.
Temperature/
humidity/ illumination
of the chamber
Specify the temperature and humidity profile inside
the chamber. Specify the light source if present.
Maximum stun-to-
stick/kill interval(s)c
Describe the maximum stun-to-stick/kill interval and
the exsanguination method (blood vessel cut) that have
been applied to guarantee unconsciousness and
insensibility of the stunned animal until the moment of
death (except for proof-of-concept studies where the
duration of unconsciousness must be determined
without sticking).
Report the stun- to-stick/kill interval(s) for the last
animal stuck that did not recover consciousness in a
group stunning situation
Calibration of the LAP
equipment and
monitoring system
Describe how the decompression procedure was
controlled and how and with which frequency the
equipment was calibrated. The monitoring equipment
should be regularly calibrated. The calibration
methods applied should be reported.
a
Referring to the legal parameter „duration of exposure‟ of other stunning methods;
b
Provide information on mean or median and range and standard deviation or interquartile range of the detailed parameter;
c
In case of simple stunning.
3.1.2. Outcome
3.1.2.1. Onset of unconsciousness and insensibility
Alternative stunning methods should disrupt the neuronal function and thereby render animals
unconscious and insensible. The extent of disruption caused by a stunning intervention and the
induction of unconsciousness and insensibility are best demonstrated using EEGs and ECoGs (EFSA,
2004, 2013d). As described in the EFSA guidance (2013c), it is acceptable that studies reporting
interventions assess the onset of unconsciousness as this state is always accompanied by the onset of
insensibility.
Animals are rendered gradually unconscious and insensible during exposure to gas mixtures, and the
animals may show signs of different stages of anaesthesia as seen in clinical veterinary practice. In
general, the different stages of anaesthesia include (1) muscle jerk (voluntary and involuntary
excitation), (2) anaesthesia (light, medium and deep), (3) respiratory and cardiovascular depression,
and finally (4) death. The stage of voluntary excitement may not be seen in animals when the
induction of unconsciousness is smooth and non-aversive. However, the rate of induction of
unconsciousness, hence the duration of different stages of anaesthesia, during exposure of animals to a
low atmosphere pressure may vary and depends mainly upon the level of oxygen tension in the
atmosphere.
In physiological terms, exposure of animals to LAPS is analogous to simulated exposure to high
altitudes and, if the partial pressure is low enough is expected to produce loss of consciousness and
sensibility via hypoxia. Hypoxia inhibits brain function, as evidenced from the gradual depression
leading to the abolition of spontaneous and evoked electrical activity. The physiological brain
mechanisms associated with the induction of unconsciousness and insensibility and the EEG
manifestations appear to be common to all terrestrial vertebrate animals. The survival time of different
regions of the brain and the spinal cord to the effects of hypoxia may vary. When animals are exposed
11. Low atmosphere pressure system for stunning poultry
EFSA Journal 2014;12(1):3488 11
to low atmosphere pressure, there is a transition period during which conscious EEG patterns change
to unconscious EEG patterns, but EEG pattern interpretation is subjective. Loss of consciousness
through hypoxia results in hyper synchronisation of the brain electrical activity as evidenced from the
appearance of slow waves (high amplitude, low frequency activity) in the EEGs of mammals, leading
to quiescent EEGs. In poultry, however, only quiescent EEGs occurred without the manifestation of
slow waves. Nevertheless, brain evoked potentials are abolished during the appearance of slow waves
in the EEGs or during the occurrence of a profoundly suppressed or quiescent EEGs. Therefore, it is
recommended that abolition of evoked electrical activity in the brain should be used as an indicator of
unconsciousness when EEG manifestations are ambiguous.
Therefore, the reliable criteria to be employed to assess LAPS in broilers during controlled laboratory
studies are (as previously reported for pigs and poultry during exposure to gas mixtures):
Profoundly suppressed or quiescent EEGs. This is indicative of a complete loss of
spontaneous brain activity or a reduction of EEG total power content to less than 10 % of the
pre-stun EEG power content, and occurs after exposure to anoxia (e.g. Raj et al., 1998;
Rodríguez et al., 2008; Llonch, 2013).
Abolition of evoked electrical activity in the brain (somatosensory evoked potentials, auditory
evoked potentials or flash visual evoked potentials), which is indicative of the brain's
incapacity to receive and process external stimuli (e.g. Raj et. al., 1997; Martoft, 2001;
Rodríguez et al., 2008).
Established stunning methods induce unique brain states that are incompatible with the persistence of
consciousness. These altered brain states are associated with certain behavioural patterns and physical
reflexes which can be used as animal-based indicators. The correlation between EEG evidence of
unconsciousness and animal-based indicators is characterized for established stunning methods,
permitting the use of animal-based indicators as proxies for unconsciousness. A list with indicators for
recognition of a successful stun in different species after exposure to hypoxic atmospheres using gas
mixtures is provided in previous EFSA opinions (EFSA, 2004, 2013d). Studies in poultry and pigs
concerning welfare suggest that loss of posture is the earliest behavioural sign of the onset of
unconsciousness. However, it may not always be possible to determine the time to loss of posture as
animals start with muscle jerks before or simultaneously they lose posture depending upon the rate of
induction of hypoxia/anoxia (Raj et al., 1997; Rodríguez et al., 2008). Other indicators of effective
stunning include dilated pupils, absence of palpebral, corneal and pupillary reflexes, apnoea, relaxed
body/lack of muscle tone and absence of response to painful stimuli such as nose pricking. In
conclusion, in studies carried out under slaughterhouse conditions, the onset and the duration of
unconsciousness and insensibility should be ascertained using the indicator that best detects
unconsciousness and that has been shown to be correlated with EEGs in laboratory experiments. If
different indicators are not in agreement, following on from the precautionary principle and to benefit
animal welfare, the one that indicates the longest time interval between application of the stunning
intervention and onset of unconsciousness should be used.
In addition to EEG evidence, partial pressure of oxygen in arterial blood and/or pulsoximetry could be
used as a direct measure of hypoxia in animals. Evidence should be provided in support of the
conclusion that the values reported are incompatible with the persistence of consciousness.
3.1.2.2. Absence of pain, distress and suffering until the loss of consciousness and sensibility
If a stunning intervention does not induce immediate unconsciousness and insensibility, the absence of
pain, distress and suffering until the onset of unconsciousness and insensibility should be assessed.
Loss of consciousness during LAPS is not immediate and animals may experience pain, distress and
suffering. At the moment, indirect animal-based measures of pain, distress and suffering have to be
used as no direct tool is available to identify them.
12. Low atmosphere pressure system for stunning poultry
EFSA Journal 2014;12(1):3488 12
Seven “groups of animal-based measures” associated with pain, distress and suffering during the
induction of unconsciousness and insensibility are presented in the guidance document (Table 9;
EFSA, 2013c): vocalisations, posture and movements, general behaviour; hormone concentrations,
blood metabolites, automatic responses and brain activity. Some research papers that describe the use
of a particular animal-based measure to assess pain, distress and suffering are included as examples,
but the list is not exhaustive. Behavioural, physiological and neurological responses to pain, distress
and suffering can be different between animals within and between species.
Animal-based measures to identify pain, distress and suffering are often subjective and have a
relatively low specificity and/or sensitivity (EFSA, 2005; Le Neindre et al., 2009). Therefore, two
criteria/rules have to be fulfilled before the LAPS is considered not to induce pain, distress and
suffering before the onset of unconsciousness and insensibility:
This means that these animal-based measures should not be significantly different between the
appropriate control and treatment groups. In this regard, in the absence of pain, distress and
suffering due to the application of the intervention, the response of animals exposed to LAPS
without decompression (control or sham operation) should not be significantly different from
the response of the animals exposed to LAPS with decompression (treatment). The possibility
that the control itself has not produced a maximum response - such that no further increases
in response could occur due to the additional pain and distress caused by the intervention -
should be demonstrated.
In general, these animal-based outcomes should be consistent at the level of the individual
animal, depending upon the species and the coping strategies (that is, consistent with respect
to their interpretation).
3.1.2.3. Duration of unconsciousness and insensibility (until death)
Council Regulation (EC) No 1099/2009 states that unconsciousness and insensibility induced by
stunning should last until the moment of death. Studies in a controlled environment should determine
the duration of unconsciousness and insensibility using EEG. Based upon the obtained results (e.g. the
shortest time to recovery of consciousness observed minus 2 standard deviations), the maximal stun-
to-stick/-kill time interval can be defined that guarantees unequivocal loss of consciousness and
sensibility until the moment of death (EFSA, 2004). The applicability of the stun-to-stick/-kill interval
should then be analysed under slaughterhouse conditions using indicators recognising recovery of
consciousness and sensibility that correlate with EEGs as established in controlled environment
studies. It is acceptable that studies on alternative stunning methods assess only the duration of
unconsciousness as this will always precede the recovery of sensibility.
In general, animals are considered to be unconscious as long as the altered brain states, as recognised
from the profound changes in EEGs that are unique to the intervention and are established to be
incompatible with the persistence of consciousness are demonstrated immediately after intervention.
When changes occurring in the spontaneous EEGs are ambiguous, abolition of evoked electrical
activity in the brain (somatosensory, visual or auditory evoked potentials) can be used as an indicator
of unconsciousness. Recovery of spontaneous or evoked electrical activity in the brain can also be
used to ascertain the time to recovery of consciousness in animals following the application of
reversible stunning. In this regard, the time to return of total EEG power content (voltage squared) to
10 % or more of the pre-stun level has been used as an indicator of recovery of consciousness (e.g. Raj
et al., 2006). The time to recovery of spontaneous activity has been reported to coincide with the time
to recovery of evoked activity in the brain (Raj and O‟Callaghan, 2004).
Indicators of recovery of consciousness after stunning are listed in EFSA scientific opinion (EFSA,
2004, 2013d), but their sequence depends on the stunning intervention. Recovery of spontaneous
breathing is considered to be the earliest indicator of recovery of consciousness, which may begin as
regular gagging (a brainstem reflex of forced/laboured breathing through the mouth) in a recumbent
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EFSA Journal 2014;12(1):3488 13
animal. These gagging movements gradually lead to resumption of rhythmic breathing. There is a lack
of information on the correlation of EEG and the sequence or the time to recovery of other indicators
of consciousness, such as pupillary, palpebral or corneal reflex. It is recommended that the indicator
that is most sensitive in detecting recovery be used. Indicators that can be measured at different stages
during slaughter can be found in EFSA (2013d).
3.2. Assessment of the eligibility criteria of the submitted studies
3.2.1. Intervention
“Physiological responses to low atmospheric pressure (LAPS) and their implications for
welfare” - McKeegan et al. (2013) (henceforth referred to as “study 1”)
The intervention is insufficiently described, making it impossible to obtain details of the variables that
are critical to an assessment of the health and welfare of the chickens during the intervention. For
example, neither the rate of decompression represented graphically (it is described as “a gradual
curve”), nor oxygen partial pressure is provided.
A continuous EEG trace, from pre-exposure until death during the LAPS intervention, is not
presented. Therefore, it is not possible to assess the changes in brain activity that occur in association
with hypoxia nor the rate of induction of unconsciousness. Neither the range nor standard deviation
corresponding to the time to onset of unconsciousness, as determined using the EEG criteria (total
power and median frequency), is reported in Study 1. This makes it impossible to characterize the
variability between birds. It is not clear how much of the changes occurring in the EEG criteria were
due to the period of darkness in the chamber as no control (i.e. sham operation) data are presented in
this study.
The conservative estimate of time to loss of consciousness is reported to be approximately 40 s, but it
is not clear whether this is the minimum, maximum or average. Nevertheless, this interpretation is not
supported by the behaviour data presented in Study 1. For example, the range of time to loss of posture
is reported to be 20 to 69 s. Loss of posture has been reported as the earliest behavioural indicator of
onset of unconsciousness during exposure to argon-induced hypoxia. Therefore, the time to onset of
unconsciousness during exposure to LAPS could be as long as 69 s. The range of time to onset of wing
flapping is reported to be 96 to 159 s. It has been suggested in previous studies on stunning of poultry
with carbon dioxide or argon-induced anoxia that wing flapping occurs as a consequence of the loss of
brain control over the spinal cord, and hence, the time to onset of wing flapping could be used as a
behavioural indicator of the time to onset of unconsciousness (Raj and Gregory, 1990; Raj et al.,
1991). Therefore, the time to loss of consciousness could be as long as 159 s after exposure to LAPS.
The absence of a clear description of the sequence of other behavioural events that might be indicative
of the reaction of chickens to the LAPS intervention, such as vocalisation, gasping, head shacking,
makes assessing the welfare consequence of the intervention impossible. Further, changes in
temperature and humidity in the chamber during the intervention were not reported.
The resting heart rate in bantam chickens was found to be 100 beats per minute and, after walking on a
treadmill at a sub-maximal speed, increased to 200 beats per minute (Green et al., 2009). The heart
rates of 400 beats per minute reported in Study 1 seem abnormally high, and probably near-maximal.
This indicates that the birds were already stressed before the intervention. The fact that the heart rate
of the birds was already near maximal before the intervention severely compromises interpretation of
the heart rate data collected during the intervention.
“A new humane method of stunning broilers using low atmospheric pressure” - Vizzier-Thaxton
et al. (2010) (henceforth referred to as “study 2”)
The intervention is insufficiently described, making it impossible to obtain details of the variables that
are critical to an assessment of the health and welfare of the chickens during the intervention. For
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EFSA Journal 2014;12(1):3488 14
example, neither the rate of decompression represented graphically (it is described as “a gradual
curve”), nor oxygen partial pressure is provided.
It is stated that behavioural observations were made for 280 s (pg. 346 of the submitted study), which
was described as the duration of the intervention in Study 1. Therefore, we must assume that the
intervention in Study 2 was the same as in Study 1 and, therefore, that the results are comparable.
It is reported that, “The first movement is associated with an awareness of a change in
atmosphere that occurs approximately 60 s after pressure is reduced. A period of head
movement begins approximately 70 s later. This is followed by wing flapping.” (pg. 346, line
7)
It is not clear whether the „awareness of the rate of change‟ or „head movement‟ could be considered
as aversive reactions in birds, and the welfare implications of these behaviours are not adequately
described.
It is reported in Table 3 that the average time to first movement was 58.7 s and the average
time from first movement to loss of posture was 64.9 s.
In this study, the average time to loss of posture was reported to be 123.6 s (58.7 + 64.9 s), which is
considerably longer than the time to loss of posture reported in Study 1.
As detailed above, there are unexplained inconsistencies between Studies 1 and 2 in terms of the
estimated times to loss of consciousness.
“The effects of low atmospheric stunning and deboning time on broiler breast meat quality” -
Schilling et al. (2012) (henceforth referred to as “study 3”)
The intervention is insufficiently described, making it impossible to obtain details of the variables that
are critical to an assessment of the health and welfare of the chickens during the intervention. For
example, the only information provided about the intervention is that the atmospheric pressure to
which the chickens were exposed was equivalent to an elevation of approximately 10 000 m and that
the chickens were inside the LAPS for 2 minutes after loss of posture and for a total of 2.5 minutes. It
is stated that additional details of the intervention are available in Cheek and Cattarazzi (2010), but
that document was not one of those submitted for assessment. No observations relating to the health or
welfare of the chickens before or during the intervention are reported.
“The Effects of Low-Atmosphere Stunning and Deboning Time on Broiler Breast Meat Quality -
Battula et al. 2008 (henceforth referred to as “study 4”)
The intervention is insufficiently described, making it impossible to obtain details of the variables that
are critical to an assessment of the health and welfare of the chickens during the intervention. For
example, the only information provided about the intervention is that the atmospheric pressure to
which the chickens were exposed was 597-632 mmHg and that they were inside the LAPS for
2 minutes. No observations relating to the health or welfare of the chickens before or during the
intervention are reported.
3.2.2. Outcome
Study 1
Based on electrocardiogram (EKG) data, cardiac arrest, and on isoelectric EEGs registered in all
experimental birds, the chickens were dead when they were removed from the LAPS. No information
on animal-based measures associated with pain, distress and suffering during the induction of
unconsciousness and insensibility were provided.
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EFSA Journal 2014;12(1):3488 15
Study 2
It is unclear from the blood analyses presented in Table 4 of Study 2 whether the cause of death was
hypoxia. This is because the authors do not state whether the blood is from the arterial or venous blood
supply, but only that it was taken from the heart. If the blood sampled was arterial, then the only
measured variables that were not in the normal range for birds are the values for PO2 (the partial
pressure of oxygen, which we assume is in units of Torr, but the units are not stated in Study 2), Hct
(haematocrit), Hgb (haemoglobin concentration), and pH. This PO2 value in the arterial blood is
definitely low enough to cause death in most bird species, particularly considering that hypoxia is
induced relatively quickly with the proposed intervention. The slightly low pH is also consistent with
the explanation that the birds died from hypoxia (it indicates a metabolic acidosis). The Hct and Hgb
values are relatively low, and indicate that the birds were anaemic. However, since the values are
similar between LAPS and electrical stunning, this is probably a characteristic of the birds themselves,
and not the stunning treatment (although it is hard to tell in the absence of a pre-stunning
measurement). The low Hct and Hgb levels would accentuate the effect of a low PO2, possibly
hastening death. It is possible that a low arterial PO2 would have been better tolerated if the birds
hadn't had such low Hct and Hgb values. This observation indicates that caution should be applied
when generalizing these findings to other poultry. Nevertheless, arterial PO2 values this low are
sufficient to kill most birds with normal Hct and Hgb values. In contrast, if the blood sampled was
venous, then it was probably not sufficiently low to cause death. This leaves the cause of death
equivocal since, if by chance venous rather than arterial blood was sampled (i.e. from the right
ventricle, which is blood that has returned from the peripheral tissues and has not yet been re-
oxygenated in the lungs), then the PO2 and pH values would reflect the deoxygenated blood supply.
The histopathological evidence reported of haemorrhagic lesions in the lungs and other organs could
be interpreted as evidence of compromised welfare in these birds during the application of the
intervention. However, overall, the information provided does not allow any conclusions to be taken
about the outcome in relation to the impact of the intervention on the welfare of the chickens.
Study 3
No information on outcome was provided beyond stating that all of the chickens were dead when they
were removed from the LAPS.
Study 4
No information on outcome was provided beyond stating that all of the chickens were dead when they
were removed from the LAPS.
4. Reporting quality
The reporting quality of a study submitted for assessment is evaluated against each of the criteria in
Table 10 (Section 4) of the guidance document (EFSA, 2013c). The decision over whether the overall
reporting quality is sufficient will be based upon the judgment of the panel experts engaged to assess
the submitted studies.
4.1. Assessment of the reporting quality of the submitted studies based on the selected
parameters
The assessed studies did not pass the eligibility assessment and, therefore, reporting quality was not
assessed (as per Figure 1).
5. Methodological quality
The methodological quality of a research study can be determined by assessing its precision and its
internal and external validity. These elements are related to the extent to which the study‟s design,
implementation, data acquisition, analysis and interpretation of results 1) minimise systematic errors
16. Low atmosphere pressure system for stunning poultry
EFSA Journal 2014;12(1):3488 16
(biases) that compromise the study‟s internal validity; 2) minimise random errors that reduce the
precision of the measurements made in the study; 3) allow broad applicability of the results beyond
any single study (= external validity). Details of the methodological quality criteria assessment are
presented in chapter 5 of the guidance document.
5.1. Quality assessment of the internal validity of the submitted studies
The assessed studies did not pass the eligibility assessment and, therefore, methodological quality was
not assessed (as per Figure 1).
5.2. The extent to which the findings are consistent with other sources of information;
A simple literature search uncovered articles published mainly by the same research groups. However,
since the rate of decompression is not fully described in the submitted studies, it was not possible to
evaluate their consistency with these other studies. For this reason, the assessment was based
exclusively on the documents submitted.
CONCLUSIONS AND RECOMMENDATIONS
CONCLUSIONS
None of the four studies meet the eligibility criteria and, therefore, were not assessed further.
The manner in which the LAPS procedure is described in the submitted studies is inconsistent and,
therefore, it was not possible to ascertain the specifics of the key parameters of the intervention nor
their impact on the welfare of the chickens. The animal welfare outcome of the intervention is
incompletely described in the submitted studies, leaving it unclear whether the rate of
decompression used in LAPS induces unconsciousness and death without causing avoidable pain
and suffering in poultry.
RECOMMENDATIONS
Researchers are advised to consult the guidance document on the process and criteria applied by
EFSA to assess studies evaluating the effectiveness of stunning interventions regarding animal
protection at the time of killing.
REFERENCES
Battula V, Schilling MW, Vizzier-Thaxton Y, Behrends JM, Williams JB and Schmidt TB, 2008. The
Effects of Low-Atmosphere Stunning and Deboning Time on Broiler Breast Meat Quality. Poultry
Science, 87, 1202-1210.
Cheek H and Cattarazzi B, inventors and assignees, 2010. United States Process Patent 7662030.
Method for humanely stunning and slaughtering poultry using controlled low atmospheric pressure.
Feb. 16, 2010.
EFSA (European Food Safety Authority), 2004. Welfare aspects of the main systems of stunning and
killing the main commercial species of animals. The EFSA Journal 2004, 45, 1–29.
EFSA (European Food Safety Authority), 2005. Aspects of the biology and welfare of animals used
for experimental and other scientific purposes. The EFSA Journal 2005, 292, 1–46.
EFSA AHAW Panel (EFSA Panel on Animal Health and Welfare), 2013a. Scientific Opinion on the
electrical parameters for the stunning of lambs and kid goats. EFSA Journal 2013;11(6):3249,
40 pp. doi:10.2903/j.efsa.2013.3249
17. Low atmosphere pressure system for stunning poultry
EFSA Journal 2014;12(1):3488 17
EFSA AHAW Panel (EFSA Panel on Animal Health and Welfare), 2013b. Scientific Opinion on the
use of carbon dioxide for stunning rabbits. EFSA Journal 2013;11(6):3250, 33 pp.
doi:10.2903/j.efsa.2013.3250
EFSA AHAW Panel (EFSA Panel on Animal Health and Welfare), 2013c. Guidance on the
assessment criteria for studies evaluating the effectiveness of stunning interventions regarding
animal protection at the time of killing. EFSA Journal 2013;11(12):3486, 41 pp. doi:
10.2903/j.efsa.2013.3486
EFSA AHAW Panel (EFSA Panel on Animal Health and Welfare), 2013d. Scientific Opinion on
monitoring procedures at slaughterhouses for poultry. EFSA Journal 2013;11(12):3521, 65 pp.
doi:10.2903/j.efsa.2013.3521
Green JA , Halsey LG, Wilson RP and Frappell PB, 2009 Estimating energy expenditure of animals
using the accelerometry technique: activity, inactivity and comparison with the heart-rate
technique. Journal of Experimental Biology, 212, 471-482.
Le Neindre PGR, Guémené D, Guichet J-L, Latouche K, Leterrier C, Levionnois OMP, Prunier A,
Serrie A and Servière J, 2009. Animal pain: identifying, understanding and minimising pain in
farm animals. Multidisciplinary scientific assessment, Summary of the expert report. INRA, Paris,
98 pp.
Llonch P, Rodriguez P, Jospin M, Dalmau A, Manteca X and Velarde A, 2013. Assessment of
unconsciousness in pigs during exposure to nitrogen and carbon dioxide mixtures. Animal, 7, 492–
498.
Martoft L, Jensen EW, Rodriguez BE, Jorgensen PF, Forslid A and Pedersen HD, 2001. Middle-
latency auditory evoked potentials during induction of thiopentone anaesthesia in pigs. Laboratory
Animals, 35, 353–363.
McKeegan DE, Sandercock DA and Gerritzen MA, 2013. Physiological responses to low atmospheric
pressure stunning and the implications for welfare. Poultry Science, 92, 858-868.
Raj ABM and Gregory NG, 1990. Effect of rate of induction of carbon dioxide anaesthesia on the time
to onset of unconsciousness and convulsions. Research in Veterinary Science, 49, 360-363.
Raj ABM, and O‟Callaghan M, 2004. Effects of electrical water bath stunning current frequencies on
the spontaneous electroencephalograms and somatosensory evoked potentials in hens. British
Poultry Science, 45: 230-236.
Raj ABM, Gregory NG and Wotton SB, 1991. Changes in the somatosensory evoked potentials and
spontaneous electroencephalogram of hens during stunning in argon-induced hypoxia. British
Veterinary Journal, 147, 322-330.
Raj ABM, Johnson SP, Wotton SB and McKinstry JL, 1997. Welfare implications of gas stunning
pigs: 3. The time to loss of somatosensory evoked potentials and spontaneous
electroencephalogram of pigs during exposure to gases. British Veterinary Journal, 153, 329-340.
Raj ABM, O‟Callaghan M and Knowles T, 2006. The effect of amount and frequency of alternating
current used in water bath stunning and neck cutting methods on spontaneous
electroencephalograms in broilers. Animal Welfare, 15, 7-18.
Raj ABM, Wotton SB, McKinstry JL, Hillebrand SJW and Pieterse C, 1998. Changes in the
somatosensory evoked potentials and spontaneous electroencephalogram of broiler chickens during
exposure to gas mixtures. British Poultry Science, 39, 686-695.
Rodriguez P, Dalmau A, Ruiz-de-la-Torre JL, Manteca X, Jensen EW, Rodriguez B, Litvan H and
Velarde A 2008. Assessment of unconsciousness during carbon dioxide stunning in pigs. Animal
Welfare, 17, 341–349.
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EFSA Journal 2014;12(1):3488 18
Schilling MW, Radhakrishnan V, Vizzier-Thaxton Y, Christensen K, Joseph P, Williams JB and
Schmidt TB, 2012. The effects of low atmosphere stunning and deboning time on broiler breast
meat quality. Poultry Science, 91, 3214-3222.
Vizzier-Thaxton Y, Christensen KD, Schilling MW, Buhr RJ and Thaxton JP, 2010. A new humane
method of stunning broilers using low atmospheric pressure. Journal of Applied Poultry Research,
19, 341-348.
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EFSA Journal 2014;12(1):3488 19
APPENDIX. ASSESSMENT OF THE ELIGIBILITY CRITERIA
Study 1: “Physiological responses to low atmospheric pressure stunning (LAPS) and implications for welfare” (McKeegan et al., 2013)
Table 2: Information provided by the submitted study in relation to the intervention
Parameter Component Comment Fulfilment criterion
(yes or no)
Animal density Animal species/ age/ type and
stocking density (number/m2
and kg
of body weight/ m2
)
Broilers chicken,
28-30 days of age,
For the rest no information was provided.
No/not provided
Duration of intervention Time to achieve the target pressures
and corresponding partial pressure of
oxygen in a single-phase system or
multi-phase system
The information reported regards only the entire LAPS cycle
from doors closed to doors opening, lasting 280 sec.
The process is not sufficiently described. There is no information
provided about the variability and duration of the intervention.
No
Rate of decompression Time/pressure treatment graphic
representation
The study reports that the decompression consists of a gradual
curve of reducing pressure over 75% of the cycle and that the
final pressure is maintained over the remaining 25% but no
information is provided if air removing grade is gradual or not. A
pressure / time profile curve is needed to assess the rate of
decompression.
The pressure achieved in this study was reported to be 20% of
ambient pressure but no supporting measurements were provided.
No information in regards to re-pressurisation.
No
Rate of changes in partial pressure of
oxygen
Time/partial pressure of oxygen
treatment graphic representation
The study refers to a final pressure maintained over the remaining
25% of the cycle and being not greater than an 80% reduction in
ambient pressure. The pressure achieved in this study should
therefore be 20% of ambient pressure.
The data are not expressed in absolute values and no information
is provided on the corresponding partial pressure of oxygen.
The corresponding partial pressure of oxygen levels indicative of
hypoxia is not provided to assess the magnitude of hypoxia.
No
Temperature/ humidity/ illumination
of the chamber
Information not provided. No
Maximum stun-to-stick/kill
interval(s)
Not applicable in this case.
The intervention reported in the study is irreversible.
Not applicable
Calibration of the LAP equipment
and monitoring system
No information is provided. No
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Table 3: Information provided by the submitted study in relation to the onset of unconsciousness and insensibility
Table 4: Information provided by the submitted study in relation to animal-based measures associated with pain, distress and suffering during the
induction of unconsciousness and insensibility
Response type Groups of animal-based
measures (ABMs)
Comments Do the ABMs suggest pain,
distress and suffering
(yes, no or not possible to assess)
Behaviour Vocalisations No information provided. Not possible to assess
Postures and movements No information provided in relation to assessment of pain, distress and suffering. Not possible to assess
General behaviour No information provided in relation to assessment of pain, distress and suffering. Not possible to assess
Physiological response Hormone concentrations No information provided. No
Blood metabolites No information provided. No
Autonomic responses Information is provided on the changes in heart rate responses but not in relation to
assessment of pain, distress and suffering.
No
Neurological response Brain activity EEG was measured but not in relation to assessment of pain, distress and suffering. No
Parameter Comment Is the induction of unconsciousness and
insensibility addressed adequately?
(yes, no or not possible to assess)
EEG EEG was measured but incomplete data was provided.
The procedures to install the electrodes and their positions are described. The method used
to derive the transformations of the EEG is reported. The sampling of the EEG signal for
analysis seems to be different for the different birds and therefore is not comparable.
The information reported therefore does not allow a full assessment of the EEG data.
Not possible to assess
Animal-based indicators to
detect onset of unconsciousness
and insensibility
The evaluation of the EEG data does not allow precise determination of the time to
unconsciousness and does not associate unconsciousness with animal-based indicators.
No
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EFSA Journal 2014;12(1):3488 21
Table 5: Information provided by the submitted study in relation to the duration of unconsciousness and insensibility or evidence of death in the case of
irreversible intervention
Parameter Comments Is the duration of unconsciousness and
insensibility addressed adequately?
(yes, no or not possible to assess)
EEG In case of irreversible stunning, it is necessary to demonstrate that the intervention kills the
animal. In this case, we need indicators of death rather than indicator of unconsciousness.
It is reported isoelectric EEG occurred in all the experimental birds before exiting the LAPS.
Yes
Animal-based indicators to
detect duration of
unconsciousness and
insensibility or death
In case of irreversible stunning, it is necessary to demonstrate that the intervention kills the
animal. It is reported that, based on EKG data, cardiac arrest occurred in all the experimental
birds before exiting the LAPS.
Yes
Study 2: “A new humane method of stunning broilers using low atmospheric pressure” (Vizzier-Thaxton et al., 2010)6
Table 6: Information provided by the submitted study in relation to the intervention
Parameter Component Comment Fulfilment criterion
(yes or no)
Animal density Animal species/ type and stocking density
(number/ m2
and kg of body weight/ m2
).
Broiler chickens
For the rest no information was
provided
No
Duration of intervention Time to achieve the target pressures and
corresponding partial pressure of oxygen in a
single-phase system or multi-phase system.
No information is provided No
Rate of decompression Time/pressure treatment graphic representation. No information is provided No
Rate of changes in partial pressure of oxygen Time/partial pressure of oxygen treatment
graphic representation.
No information is provided No
Temperature humidity/ illumination of the chamber No information is provided No
Maximum stun-to-stick/kill interval(s) Not applicable in this case
The intervention reported in the study
is irreversible
Not applicable
Calibration of the LAP equipment and monitoring
system
No detailed information is provided No
6
(The assessment of this study only focuses on the intervention of low atmospheric pressure and not on the electrical stunning intervention)
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Table 7: Information provided by the submitted study in relation to the onset of unconsciousness and insensibility
Parameter Comment Is the induction of unconsciousness and
insensibility addressed adequately?
(yes, no or not possible to assess)
EEG Not measured. No
Animal-based indicators to detect
onset of unconsciousness and
insensibility
Some indicators are used but the purpose is not clear. In particular, the use of loss of
posture as indicator to assess onset of unconsciousness is not properly described.
Onset of wing flapping could be used as indicators of onset of unconsciousness, but
wing flapping were not interpreted in that context.
No
Table 8: Information provided by the submitted study in relation to animal-based measures associated with pain, distress and suffering during the
induction of unconsciousness and insensibility
Response type Groups of animal-based
measures (ABMs)
Comment Do the ABMs suggest pain, distress and
suffering (yes, no or not possible to assess)
Behaviour Vocalisations No information provided. No
Postures and movements Information provided is not sufficient to interpret the use of the
indicators in relation to assessment of pain, distress and suffering.
Not possible to assess
General behaviour Information provided is not sufficient to interpret the use of the
indicators in relation to assessment of pain, distress and suffering.
Not possible to assess
Physiological response Hormone concentrations Corticosterone concentration was not used to assess the welfare of
birds during decompression. It was reported only in comparison to
electrically stunned broilers and it was no compared to a baseline
control.
No
Blood metabolites No metabolites indicators of stress were measured. No
Autonomic responses No information provided. No
Neurological response Brain activity Not measured. No
Table 9: Information provided by the submitted study in relation to the duration of unconsciousness and insensibility or evidence of death in the case of
irreversible intervention
Parameter Comment Is the duration of unconsciousness and
insensibility addressed adequately?
(yes, no or not possible to assess)
EEG Not measured No
Animal-based indicators to detect duration
of unconsciousness and insensibility
In case of irreversible stunning, it is necessary to demonstrate that the
intervention kills the animal. No indicator of death is used.
No
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EFSA Journal 2014;12(1):3488 23
Study 3: The effects of low atmosphere stunning and deboning time on broiler breast meat quality (Schilling et al., 2012)
Table 10: Information provided by the submitted study in relation to the intervention
Parameter Component Comment Fulfilment
criterion
(yes or no)
Animal density Animal species/ age/ type
and stocking density
(number/m2
and kg of body
weight/ m2
)
In the described study, 2 full live haul cages with 250 commercial broilers each, were
inserted in the chamber. No information is provided on the stocking density in
cages/chamber.
No
Duration of intervention Time to achieve the target
pressures and corresponding
partial pressure of oxygen in
a single-phase system or
multi-phase system
Information is reported only on the total time during which broilers were kept in the
container (2.5 min). Corresponding partial pressure of oxygen is not provided.
All broilers were maintained in the LAPS chamber for 2 min after loss of posture for a total
time of 2.5 min in the container where the pressure was reduced to that at an approximate
elevation of 10,000 m.
No
Rate of decompression Time/pressure treatment
graphic representation
The study refers to another publication illustrating the low atmospheric pressures created in
LAPS (in the range of 150 to 230 mmHg) and rate of change from sea level. It also refers to
the commercial prototype that was used (Technocatch LLC, Kosciusko, MS). Information is
given on the type of vacuum pumps rated at 14 m3/
min.
No
Rate of changes in partial
pressure of oxygen
Time/partial pressure of
oxygen treatment graphic
representation
No information is provided. No
Temperature/ humidity/
illumination of the chamber
No information is provided. No
Maximum stun-to-stick/kill
interval(s)
No information is provided. No
Calibration of the LAP
equipment and monitoring
system
A computer-based data acquisition and control system was used to monitor tank pressure and
control pump action but data are not provided. No information is provided on the calibration
of the system.
No
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Table 11: Information provided by the submitted study in relation to the onset of unconsciousness and insensibility
Parameter Comment
This study is not relevant to the assessment of impact on health and welfare
Is the induction of unconsciousness and
insensibility addressed adequately?
(yes, no or not possible to assess)
EEG No information is provided. no
Animal-based indicators to detect onset of
unconsciousness and insensibility
Time to onset of unconsciousness is not provided. no
Table 12: Information provided by the submitted study in relation to animal-based measures associated with pain, distress and suffering during the
induction of unconsciousness and insensibility
Response type Groups of animal-based
measures (ABMs)
Comments Do the ABMs suggest pain,
distress and suffering
(yes, no or not possible to assess)
Behaviour Vocalisations No information is provided. Not possible to assess
Postures and movements It is reported that broilers are maintained in the LAPS chamber for 2 min
after loss of posture but no data are provided.
Not possible to assess
General behaviour No information is provided. Not possible to assess
Physiological response Hormone concentrations No information is provided. Not possible to assess
Blood metabolites No information is provided. Not possible to assess
Autonomic responses No information is provided. Not possible to assess
Neurological response Brain activity No information is provided. Not possible to assess
Table 13: Information provided by the submitted study in relation to the duration of unconsciousness and insensibility or evidence of death in the case of
irreversible intervention
Parameter Comments
This study is not relevant to the assessment of impact on health and welfare
Is the duration of unconsciousness and
insensibility addressed adequately?
(yes, no or not possible to assess)
EEG No information is provided. Not possible to assess
Animal-based indicators to detect duration
of unconsciousness and insensibility and
death
No information is provided. Not possible to assess
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Study 4: The Effects of Low-Atmosphere Stunning and Deboning Time on Broiler Breast Meat Quality (Battula et al., 2008)
Table 14: Information provided by the submitted study in relation to the intervention
Parameter Component Comments Fulfilment criterion
(yes or no)
Animal density Animal species/ age/ type and stocking
density (number/m2
and kg of body
weight/ m2
)
The study reports that cages with 24 broilers were placed into the
chamber but the stocking density in cages/chamber is not reported.
No
Duration of intervention Time to achieve the target pressures
and corresponding partial pressure of
oxygen in a single-phase system or
multi-phase system
Information is provided on the time during which broilers were kept in
decompression chamber (2 min) but corresponding partial pressure of
oxygen is not provided. It is not specified if decompression starts as soon
as door closes.
No
Rate of decompression Time/pressure treatment graphic
representation
The study refers to the commercial prototype that was used (Technocatch
LLC, Kosciusko, MS). Information is given on the type of vacuum
pumps rated at 14 m3
/min. The low atmospheric pressure achieved in the
chamber was described as being 597 to 632 mmHg.
No
Rate of changes in
partial pressure of
oxygen
Time/partial pressure of oxygen
treatment graphic representation
No information is provided. No
Temperature/ humidity/
illumination of the
chamber
No information provided. No
Maximum stun-to-
stick/kill interval(s)
Information not provided; the study only reports that after ataxia, broilers
were decapitated manually.
No
Calibration of the LAP
equipment and
monitoring
A computer-based data acquisition and control system was used to
monitor tank pressure and control pump action. No information is
provided on the calibration of the system.
No
Table 15: Information provided by the submitted study in relation to the onset of unconsciousness and insensibility
Parameter Comments
This study is not relevant to the assessment of impact on health and welfare
Is the induction of unconsciousness and
insensibility addressed adequately?
(yes, no or not possible to assess)
EEG No information provided. no
Animal-based indicators to detect onset of
unconsciousness and insensibility
Time to onset of unconsciousness is not provided. no
26. Low atmosphere pressure system for stunning poultry
EFSA Journal 2014;12(1):3488 26
Table 16: Information provided by the submitted study in relation to animal-based measures associated with pain, distress and suffering during the
induction of unconsciousness and insensibility
Response type Groups of animal-based
measures (ABMs)
Comments Do the ABMs suggest pain, distress
and suffering
(yes, no or not possible to assess)
Behaviour Vocalisations No information is provided. Not possible to assess
Postures and movements It is reported that after ataxia (loss of posture, resulting in the inability to
maintain a standing position and no neck tension at the onset of
unconsciousness), broilers were decapitated manually but no information that
would permit an assessment of pain, distress and suffering is provided.
Not possible to assess
General behaviour No information is provided. Not possible to assess
Physiological response Hormone concentrations No information is provided. Not possible to assess
Blood metabolites No information is provided. Not possible to assess
Autonomic responses No information is provided. Not possible to assess
Neurological response Brain activity No information is provided. Not possible to assess
Table 17: Information provided by the submitted study in relation to the duration of unconsciousness and insensibility or evidence of death in the case of
irreversible intervention
Parameter Comment Is the duration of unconsciousness/insensibility
addressed adequately?
(yes, no or not possible to assess)
EEG No information is provided. Not possible to assess
Animal-based indicators to detect duration of
unconsciousness/ insensibility or death
No information is provided. Not possible to assess
27. Low atmosphere pressure system for stunning poultry
EFSA Journal 2014;12(1):3488 27
GLOSSARY AND ABBREVIATIONS
ABBREVIATIONS
ABM Animal-based measure
AHAW Panel EFSA Panel on Animal Health and Welfare
EC European Commission
EFSA European Food Safety Authority
ECoG Electrocorticogram
EEG Electroencephalogram
EKG Electrocardiogram
Hct Hematocrit
Hgb Haemoglobin concentration
LAPS Low atmosphere pressure system