Target Animals Research Articles

A non‐invasive method for sampling the body odour of mammals

Abstract Olfaction is a central aspect of mammalian communication, providing information about individual attributes such as identity, sex, group membership or genetic quality. Yet, the chemical underpinnings of olfactory cues remain little understood, one of the reasons being the difficulty in obtaining high quality samples for chemical analysis. In this study, we adjusted and evaluated the use of thermal desorption (TD) tubes, commonly used in plant metabolomic and environmental studies, for non‐invasive sampling of mammalian body odour. We obtained chemical profiles of meerkat (Suricata suricatta) body odour samples, using TD tubes analysed with gas chromatography–mass spectrometry. TD tubes captured a wide range of volatile and semi‐volatile organic compounds, including compounds likely originating from the target animals. Adjustment of sampling parameters (distance, volume, flow rate, interruption of sampling) to increase the feasibility for a non‐invasive application yielded samples of adequate quality. However, to minimize the variability between samples, sampling parameters should be kept constant and samples should be collected when no conspecifics are close‐by. The method was sensitive enough to pick up population differences in the chemical profiles of two captive groups of meerkats, demonstrating its applicability to biological questions. With sufficiently habituated animals, the method is applicable non‐invasively, allowing short‐ and long‐term studies on a wide range of questions, including e.g. chemical signatures of kinship, diet, individual health or reproductive state.

Genotype imputation in a tropical crossbred dairy cattle population

The objective of this study was to investigate different strategies for genotype imputation in a population of crossbred Girolando (Gyr × Holstein) dairy cattle. The data set consisted of 478 Girolando, 583 Gyr, and 1,198 Holstein sires genotyped at high density with the Illumina BovineHD (Illumina, San Diego, CA) panel, which includes ∼777K markers. The accuracy of imputation from low (20K) and medium densities (50K and 70K) to the HD panel density and from low to 50K density were investigated. Seven scenarios using different reference populations (RPop) considering Girolando, Gyr, and Holstein breeds separately or combinations of animals of these breeds were tested for imputing genotypes of 166 randomly chosen Girolando animals. The population genotype imputation were performed using FImpute. Imputation accuracy was measured as the correlation between observed and imputed genotypes (CORR) and also as the proportion of genotypes that were imputed correctly (CR). This is the first paper on imputation accuracy in a Girolando population. The sample-specific imputation accuracies ranged from 0.38 to 0.97 (CORR) and from 0.49 to 0.96 (CR) imputing from low and medium densities to HD, and 0.41 to 0.95 (CORR) and from 0.50 to 0.94 (CR) for imputation from 20K to 50K. The CORRanim exceeded 0.96 (for 50K and 70K panels) when only Girolando animals were included in RPop (S1). We found smaller CORRanim when Gyr (S2) was used instead of Holstein (S3) as RPop. The same behavior was observed between S4 (Gyr + Girolando) and S5 (Holstein + Girolando) because the target animals were more related to the Holstein population than to the Gyr population. The highest imputation accuracies were observed for scenarios including Girolando animals in the reference population, whereas using only Gyr animals resulted in low imputation accuracies, suggesting that the haplotypes segregating in the Girolando population had a greater effect on accuracy than the purebred haplotypes. All chromosomes had similar imputation accuracies (CORRsnp) within each scenario. Crossbred animals (Girolando) must be included in the reference population to provide the best imputation accuracies.

Safety and efficacy of VevoVitall® (benzoic acid) as feed additive for minor porcine species.

Following a request from the European Commission, the EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) was asked to deliver a scientific opinion on VevoVitall® (benzoic acid) as a feed additive for minor porcine species, when used as zootechnical additive at a maximum level of 5,000 mg/kg complete feed. The safety and efficacy of the additive had been evaluated by the FEEDAP Panel on weaned piglets (at the maximum supplementation level of 5,000 mg/kg complete feed) and pigs for fattening and pigs for reproduction (at the maximum level of 10,000 mg/kg complete feed). Subsequent to EFSA's assessments, the additive has been authorised for use in feed of weaned piglets, pigs for fattening and sows. To the knowledge of the FEEDAP Panel, there was no new information that might modify its previous assessments on VevoVitall® concerning safety for consumers and the environment; therefore, this opinion focused only on the safety of the target animals and the efficacy of the product. The FEEDAP Panel concluded that VevoVitall® is safe at the supplementation level of 5,000 mg/kg complete feed for minor porcine species for fattening and for reproduction; the FEEDAP could not conclude on the safety of the additive for weaned minor porcine species. VevoVitall® does not represent a risk for the consumer and the environment. VevoVitall® does not pose a risk by inhalation to users and is not skin sensitiser, but is a skin irritant and a severe eye irritant. VevoVitall® has the potential to decrease the urinary pH in minor porcine species at the dose of 5,000 mg/kg complete feed. A remark on the relevance of the claim has been posted by the Panel.

Pan-neuronal calcium imaging with cellular resolution in freely swimming zebrafish.

Calcium imaging with cellular resolution typically requires an animal to be tethered under a microscope, which substantially restricts the range of behaviors that can be studied. To expand the behavioral repertoire amenable to imaging, we have developed a tracking microscope that enables whole-brain calcium imaging with cellular resolution in freely swimming larval zebrafish. This microscope uses infrared imaging to track a target animal in a behavior arena. On the basis of the predicted trajectory of the animal, we applied optimal control theory to a motorized stage system to cancel brain motion in three dimensions. We combined this motion-cancellation system with differential illumination focal filtering, a variant of HiLo microscopy, which enabled us to image the brain of a freely swimming larval zebrafish for more than an hour. This work expands the repertoire of natural behaviors that can be studied with cellular-resolution calcium imaging to potentially include spatial navigation, social behavior, feeding and reward.

Hand feeding can periodically fuel a major portion of bull shark energy requirements at a provisioning site in Fiji

AbstractWildlife tourism is often extolled for its contribution to conservation. However, understanding the effects of tourism activities on the health of target animals is required to fully assess conservation benefits. Shark tourism operators often use food rewards to attract sharks in close proximity to tourists, but nothing is known about the contribution of these food rewards to the energetic requirements of target species. In this study, hand feeding of bull sharks Carcharhinus leucas was directly observed on 36 commercial shark watching dives in the Shark Reef Marine Reserve (SRMR), Fiji. Mean number of tuna heads consumed per dive by focal individuals ranged from 1.3 to 3.7. Monitored bull sharks consumed an average of ~0.74 heads per provisioning day, and bioenergetics modelling suggests that some sharks might periodically be meeting their full energy requirement from provisioning at the SRMR. Knowing how much individual sharks consume at provisioning sites and how this relates to their energy requirements is crucial in order to better understand the effects of wildlife tourism and its contribution to conservation.

Health effects of feeding genetically modified (GM) crops to livestock animals: A review

A large share of genetically modified (GM) crops grown worldwide is processed into livestock feed. Feed safety of GM crops is primarily based on compositional equivalence with near-isogenic cultivars and experimental trials in rodents. However, feeding studies in target animals add to the evaluation of GM crops with respect to animal health. This review aimed to evaluate the possible health effects of feeding GM crops to livestock by reviewing scientific publications on experimental studies in ruminants, pigs, and poultry in which at least one of the following health parameters was investigated: body condition score, organ weight, haematology, serum biochemistry, histopathology, clinical examination, immune response, or gastrointestinal microbiota. In most experiments, either Bt (Bacillus thuringiensis) maize, Roundup Ready (RR) soybean, or both were fed to livestock animals. Significant differences (P<0.05) in health parameters were most often observed when animals were fed Bt maize, although most effects measured were unlikely to be of biological significance and were within normal biological ranges. Health effects of RR soybean were only observed in one experimental study with broilers. Based on this literature review, we conclude that there is no clear evidence that feed composed of first generation GM crops has adverse effects on animal health.

Safety of Strontium Chloride as a Skeletal Marking Agent for Pacific Salmon.

The purpose of this study was to evaluate the biological effects associated with administering strontium chloride as a marking agent to age-0 Chinook Salmon Oncorhynchus tshawytscha fry. Fish were held in a 0× (0 mg/L), 1× (3,000 mg/L; current standard dosage), 3× (9,000 mg/L), or 5× (15,000 mg/L) solution of strontium chloride for 72 h (three times the standard duration of 24 h). The mortality among fish in the 5× strontium chloride exposure group was significantly higher than that observed in the other groups. A dose-related effect on general fish behavior and on feeding behavior was observed. Fish in all test tanks appeared to feed to satiation, except for fish in the 5× tanks during days 2 and 3. Fish in all other test tanks behaved normally. No dose-related effect on fish growth was detected. Histopathological evaluations showed that fish in the 5× exposure group had a significantly higher number of gill lesions than the 0× group. Our mortality, behavioral, and histological assessments suggested that juvenile Chinook Salmon could be safely immersed for three consecutive days in a 9,000-mg/L solution of strontium chloride. This finding potentially expands the present 1,000-3,000-mg/L dosage and 24-h holding period that can be used to mark juvenile fish with strontium chloride solutions. The research also provides necessary target animal safety data for U.S. Food and Drug Administration approval of strontium chloride as an alternative marking method that is suitable for fish with a short holding time. Received February 19, 2017; accepted July 16, 2017.

Safety and nutritional value of a dried killed bacterial biomass from Escherichiacoli (FERM BP-10942) (PT73 (TM)) as a feed material for pigs, ruminants and salmonids.

PT73 (TM) is a dried, heat‐inactivated bacterial biomass used as a feed material produced from an Escherichia coli K‐12 strain, which was genetically modified to overproduce threonine. The recipient organism E. coli MG 1655 is considered to be safe. The traits introduced in the final modified strain E. coli FERM BP‐10942 are mainly limited to the overproduction of threonine. No full‐length antibiotic resistance genes or other sequences of concern remain in the modified strain. In conclusion, the EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP): does not identify risks for human and animal health or the environment from this biomass regarding the genetic modification of the strain. The proposed recommended use level for dairy cows (8% PT73 (TM) of feed dry matter (~ 7% in complete feed)) and salmonids (13%) is considered safe for these target animals. The conclusion form dairy cows could be extended to other ruminants (from the beginning of rumination). Complete feed for pigs for fattening may contain up to 10% PT73 (TM). The toxicological data indicate effects of PT73 (TM) on blood coagulation and liver, which are considered to be adverse. As a consequence, the FEEDAP Panel is unable to conclude on the safety for the consumer of products derived from animals receiving feed containing PT73 (TM). PT73 (TM) is not considered a skin/eye irritant but should be considered as a potential skin and respiratory sensitiser. Moreover, any exposure of users to dust from the product via the inhalation route should be considered a serious risk. The FEEDAP Panel considers that substitution of PT73 (TM) for other protein‐rich feed materials will not adversely affect the environment.

Example-based expressive animation of 2D rigid bodies

We present a novel approach to facilitate the creation of stylized 2D rigid body animations. Our approach can handle multiple rigid objects following complex physically-simulated trajectories with collisions, while retaining a unique artistic style directly specified by the user. Starting with an existing target animation (e.g., produced by a physical simulation engine) an artist interactively draws over a sparse set of frames, and the desired appearance and motion stylization is automatically propagated to the rest of the sequence. The stylization process may also be performed in an off-line batch process from a small set of drawn sequences. To achieve these goals, we combine parametric deformation synthesis that generalizes and reuses hand-drawn exemplars, with non-parametric techniques that enhance the hand-drawn appearance of the synthesized sequence. We demonstrate the potential of our method on various complex rigid body animations which are created with an expressive hand-drawn look using notably less manual interventions as compared to traditional techniques.

Biochemical changes and drug residues in ascidian Halocynthia roretzi after formalin\u2013hydrogen peroxide treatment regimen designed against soft tunic syndrome

Soft tunic syndrome (STS) is a protozoal disease caused by Azumiobodo hoyamushi in the edible ascidian Halocynthia roretzi. Previous studies have proven that combined formalin–hydrogen peroxide (H2O2) bath is effective in reducing STS progress and mortality. To secure target animal safety for field applications, toxicity of the treatment needs to be evaluated. Healthy ascidians were bathed for 1 week, 1 h a day at various bathing concentrations. Bathing with 5- and 10-fold optimum concentration caused 100% mortality of ascidians, whereas mortality by 0.5- to 2.0-fold solutions was not different from that of control. Of the oxidative damage parameters, MDA levels did not change after 0.5- and 1.0-fold bathing. However, free radical scavenging ability and reducing power were significantly decreased even with the lower-than-optimal 0.5-fold concentration. Glycogen content tended to increase with 1-fold bathing without statistical significance. All changes induced by the 2-fold bathing were completely or partially restored to control levels 48 h post-bathing. Free amino acid analysis revealed a concentration-dependent decline in aspartic acid and cysteine levels. In contrast, alanine and valine levels increased after the 2-fold bath treatment. These data indicate that the currently established effective disinfectant regimen against the parasitic pathogen is generally safe, and the biochemical changes observed are transient, lasting approximately 48 h at most. Low levels of formalin and H2O2 were detectable 1 h post-bathing; however, the compounds were completely undetectable after 48 h of bathing. Formalin–H2O2 bathing is effective against STS; however, reasonable care is required in the treatment to avoid unwanted toxicity. Drug residues do not present a concern for consumer safety.

Efficacy of Levucell® SB (Saccharomyces cerevisiae CNCM I-1079) as a feed additive for weaned piglets.

Levucell® SB is a feed additive based on viable cells of a strain Saccharomyces cerevisiae marketed in three formulations: SB20 with a minimum concentration of 2 × 1010 CFU/g viable yeast cells and SB10 ME and SB Titan, both microencapsulated forms with a minimum concentration of 1 × 1010 CFU/g viable yeast cells. In 2014, EFSA was requested by the European Commission to re‐evaluate the product when used as a zootechnical additive in feed for sows and weaned piglets. The safety of the additive for consumers, users, the environment and target animals and its efficacy for sows was established at that time. In that instance, the FEEDAP also concluded on the potential Levucell® SB to improve growth of piglets based on one study where supplementation of the additive was done at the recommended dose of 2 × 109 CFU/kg feedingstuffs and two at 1 × 109 CFU/kg feedingstuffs, and considered that the effective dose was 2 × 109 CFU/kg feedingstuffs. Two additional feeding trials in weaned piglets receiving the additive at 1 × 109 CFU/kg feedingstuffs are the subject of this assessment. Due to the high mortality, one of the studies was not considered. In the remaining study, the supplementation of Levucell® SB to weaned piglets at 1 × 109 CFU/kg feed resulted in a significant improvement of feed to gain ratio. Taking into account the two trials from the initial application which showed significant effects at 1 × 109 CFU/kg feed, the FEEDAP Panel concludes that Levucell® SB has the potential to improve performance of weaned piglets when added to feed at 1 × 109 CFU/kg complete feedingstuffs. Results of the efficacy studies can be applied to any of the three available formulations when used to deliver the same dose.

Safety and efficacy of Levucell®SC (Saccharomyces cerevisiae CNCM I-1077) as a feed additive for dairy cows, cattle for fattening, minor ruminant species and camelids.

Following a request from the European Commission, the EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) was asked to deliver a scientific opinion on the safety and efficacy of Levucell® SC for dairy cows, cattle for fattening, all minor ruminant species and camelids. The additive consists of viable cells of a strain of Saccharomyces cerevisae and is sold in three formulations. The FEEDAP Panel considers that the three available formulations are equivalent when used to deliver the same dose. The active agent fulfils the requirements of the qualified presumption of safety approach to the assessment of safety and since no concerns are expected from other components, Levucell® SC can be presumed safe for target animals, consumers of products from treated animals and the environment. Levucell® SC is not a skin irritant or sensitiser but is an eye irritant. Inhalation exposure is unlikely. Encapsulation is not expected to introduce hazards for users. The FEEDAP Panel is not in the position to conclude on the efficacy of Levucell® SC for dairy cows, minor dairy ruminant species or dairy camelids. Levucell® SC has a potential to improve the performance of cattle raised for fattening when supplied at a minimum dose of 8 × 109 CFU/head and day which would approximate to 6 × 108 CFU/kg complete feed. This conclusion is extended to minor ruminant species and camelids reared for meat production at the same minimum dose of 6 × 108 CFU/kg complete feed.

Safety and efficacy of selenium-enriched yeast (Saccharomycescerevisiae CNCM I-3399) for all animal species.

Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the safety and efficacy of selenium‐enriched yeast (Saccharomyces cerevisiae CNCM I‐3399) for all animal species, based on a dossier submitted for the modification of the terms of authorisation of the additive. The additive is currently authorised as selenomethionine produced by S. cerevisiae CNCM I‐3399 as a nutritional additive (compound of trace elements) with a minimum selenium content of 2,000 mg/kg. The applicant proposed the inclusion of an additional formulation with a minimum content of selenium in the additive of 3,000 mg/kg. Considering (i) that there are no relevant changes in the manufacturing of the product compared to the former application and (ii) that the conditions of use already authorised remain the same, the EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) stated that the modification requested would not affect the conclusions of the FEEDAP Panel on safety for the consumer, safety for the environment and efficacy of the product made in a previous assessment of a similar product with a lower selenium concentration. Since the capacity of the additive to homogeneously distribute in feed was proven, the tolerance studies already provided for the currently authorised product could be used to conclude on the safety of the additive for the target animals. Selenium is hazardous upon inhalation; owing to the dusting potential and the selenium content of dust, persons handling the additive are at risk. The additive should be considered as a respiratory sensitiser. The additive is not an irritant for eyes and skin. No conclusions can be reached on the dermal sensitising properties of the additive.

Modern science for better quality control of medicinal products “Towards global harmonization of 3Rs in biologicals”: The report of an EPAA workshop

This article summarizes the outcome of an international workshop organized by the European Partnership for Alternative Approaches to Animal Testing (EPAA) on Modern science for better quality control of medicinal products: Towards global harmonization of 3Rs in biologicals. As regards the safety testing of biologicals, the workshop participants agreed to actively encourage the deletion of abnormal toxicity tests and target animal batch safety tests from all relevant legal requirements and guidance documents (country-specific guidelines, pharmacopoeia monographs, WHO recommendations). To facilitate the global regulatory acceptance of non-animal methods for the potency testing of, e.g., human diphtheria and tetanus vaccines and veterinary swine erysipelas vaccines, international convergence on the scientific principles of the use of appropriately validated in vitro assays for replacing in vivo methods was identified as an overarching goal. The establishment of scientific requirements for new assays was recognized as a further means to unify regulatory approaches in different jurisdictions. It was recommended to include key regulators and manufacturers early in the corresponding discussions. Manufacturers and responsible expert groups, e.g. at the European Directorate for the Quality of Medicines and Health Care of the Council of Europe or the European Medicines Agency, were invited to consider leadership for international collaboration.

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

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

Hematologic RIs for healthy water buffaloes (Bubalus bubalis) in southern Mexico.

Baseline hematologic data are used by veterinarians and wildlife biologists to perform health assessments on target animal species. Hematologic measurements are influenced by various factors including geography. Baseline hematologic RIs have been established for domestic water buffaloes (Bubalus bubalis) from various countries in the Eastern Hemisphere, but these data are not readily available for the Western Hemisphere. The aim of this study was to determine hematologic values for domestic water buffaloes from several commercial farms in southern Mexico. Blood was collected from 126 healthy, postlactating females (3 to 10 years old) from the Murrah breed, and 10 hematologic variables were measured. Means, SDs, RIs, medians (MED), median absolute deviations (MAD), and other statistics were calculated for each hematologic variable. The MED (and MAD) for each variable are as follows: RBC count, 7.6 (1.1) × 1012 /L; hemoglobin, 116.0 (13.3) g/L; PCV, 41.5 (7.6) %; MCV, 56.8 (7.0) fL; MCH, 14.6 (1.6) pg; MCHC, 250.0 (35.6) g/L; RDW (SD), 29.7 (5.5) fL; RDW (CV), 18.2 (1.4) %; reticulocytes, 0.0 (0.0) %, and WBC count, 12.4 (1.3) × 109 /L. These values were compared to those previously reported for water buffaloes from several countries in the Eastern Hemisphere and, on most occasions, they differed significantly. Our data can be used by veterinarians and other personnel involved in buffalo production in Mexico during medical evaluations.

Safety and efficacy of Bacillusamyloliquefaciens (NCIMB30229) as a silage additive for all animal species.

The additive under assessment is a preparation of Bacillus amyloliquefaciens NCIMB 3022 intended for use as a technological additive in forages for all animal species at a proposed minimum dose of 5 × 107 CFU/kg fresh materials. The species B. amyloliquefaciens is considered by EFSA to be suitable for the qualified presumption of safety approach to safety assessment. This approach requires the identification of the strain and evidence that it is not toxigenic and does not show acquired resistance to relevant antibiotics. In a previous assessment, the identity and susceptibility to clinically relevant antibiotics of the active agent was established but the lack of toxigenic potential could not be demonstrated. Therefore, the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) could not conclude on the safety of the product for target animals, consumers, users or the environment. In the same opinion, the potential of the additive to improve aerobic stability of silages at the proposed dose was not convincingly demonstrated. A new cytotoxicity study was conducted using methanol extracts. The methanol extract was cytotoxic at the inclusion level of 5%, while no cytotoxicity was observed at lower concentrations. The current guidance on Bacillus requires absence of toxic effect using the non‐concentrated supernatant. Since the applicant was unable to determine the correlation between the methanol extracts and the Bacillus supernatant concentrations, the FEEDAP Panel is unable to conclude on the toxigenic potential of the strain based on the current data. An additional efficacy study with a similar protocol to the ones previously assessed was conducted. The study showed an improved aerobic stability when added to forage at the proposed dose. However, the FEEDAP Panel remains unable to conclude on the efficacy of the additive to improve the ensiling process on the basis of a single positive result.

Phytase produced using Schizosaccharomyces pombe ASP595-1 strain (Genetically Modified Feed Additives).

Food Safety Commission of Japan (FSCJ) conducted two sets of risk assessment on phytase produced using Schizosaccharomyces pombe ASP595-1 strain. One is the safety assessment on feed additives produced using genetically modified microorganisms. None of new harmful substance is generated in the additive and thus not transferred to animal products, such as meat, milk and eggs. Components associated with genetic modification in the additive are unlikely to yield harmful substances to be accumulated in animal products. The components are unlikely to affect the metabolism of the target animals to generate new harmful substances. No safety concern is thus raised in the products from animals fed with this phytase. The other is the risk assessment related to the revision of the Standards and Specifications of Feeds and Feed Additives. The feed additive was considered to have no genotoxicity relevant to human health. No adverse effects were observed in 14-day and 90-day subacute toxicity studies in rats as well as in feeding trials in pigs and poultry. On the formulations with polyvinyl alcohol, human intake of polyvinyl alcohol via animal products was recognized to be negligible. Consequently, FSCJ judged that the risk on human health via food is negligible as long as the 6-phytase produced using S. pombe ASP595-1 strain is properly used as a feed additive.

Safety and efficacy of zinc chelate of methionine sulfate for all animal species

The additive ‘Zinc chelate of methionine sulfate’ is zinc chelated with methionine in a molar ratio 1:1, with a minimum zinc content of 19.1%. Owing to the limitations of the tolerance study, it could not be used for the assessment of safety for target animals; therefore, the EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP Panel) cannot conclude on the safety of zinc chelate of methionine sulfate for the target species. No concerns for consumer safety are expected from the use of the zinc chelate of methionine sulfate in animal nutrition when used up to the maximum EU authorised zinc levels in feed. Zinc chelate of methionine sulfate should be considered as a skin and eye irritant, and as a skin sensitiser; it is considered to pose a risk by inhalation to the users. The additive under assessment, zinc chelate of methionine sulfate, is intended to be a substitute for other authorised zinc additives and will not further increase the environmental burden of zinc; therefore, the FEEDAP Panel considers that the use of the additive in animal nutrition would not pose an additional risk for the environment. Based on literature studies and a specific study conducted with the additive under assessment, zinc chelate of methionine sulfate is an available source of zinc for all animal species.

Surface reflectance drives nest box temperature profiles and thermal suitability for target wildlife.

Thermal properties of tree hollows play a major role in survival and reproduction of hollow-dependent fauna. Artificial hollows (nest boxes) are increasingly being used to supplement the loss of natural hollows; however, the factors that drive nest box thermal profiles have received surprisingly little attention. We investigated how differences in surface reflectance influenced temperature profiles of nest boxes painted three different colors (dark-green, light-green, and white: total solar reflectance 5.9%, 64.4%, and 90.3% respectively) using boxes designed for three groups of mammals: insectivorous bats, marsupial gliders and brushtail possums. Across the three different box designs, dark-green (low reflectance) boxes experienced the highest average and maximum daytime temperatures, had the greatest magnitude of variation in daytime temperatures within the box, and were consistently substantially warmer than light-green boxes (medium reflectance), white boxes (high reflectance), and ambient air temperatures. Results from biophysical model simulations demonstrated that variation in diurnal temperature profiles generated by painting boxes either high or low reflectance colors could have significant ecophysiological consequences for animals occupying boxes, with animals in dark-green boxes at high risk of acute heat-stress and dehydration during extreme heat events. Conversely in cold weather, our modelling indicated that there are higher cumulative energy costs for mammals, particularly smaller animals, occupying light-green boxes. Given their widespread use as a conservation tool, we suggest that before boxes are installed, consideration should be given to the effect of color on nest box temperature profiles, and the resultant thermal suitability of boxes for wildlife, particularly during extremes in weather. Managers of nest box programs should consider using several different colors and installing boxes across a range of both orientations and shade profiles (i.e., levels of canopy cover), to ensure target animals have access to artificial hollows with a broad range of thermal profiles, and can therefore choose boxes with optimal thermal conditions across different seasons.