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Rabies causes approximately 20,000 human deaths in India each year. Nearly all of these occur following dog bites. Large-scale, high-coverage dog rabies vaccination campaigns are the cornerstone of rabies elimination strategies in both human and dog populations, although this is particularly challenging to achieve in India as a large proportion of the dog population are free-roaming and unowned. Further, little is known about free-roaming dog ecology in India which makes defining optimum vaccination strategies difficult. In this study, data collected using a mobile phone application during three annual mass vaccination and neutering (surgical sterilisation of both males and females) campaigns of free-roaming dogs in Ranchi, India (during which a total of 43,847 vaccinations, 26,213 neuter surgeries and 28,172 re-sight observations were made) were interrogated, using two novel approaches to estimate the proportion of neutered dogs that were lost from the city (assumed due to mortality or migration) between campaign years. Analysis revealed high losses of neutered dogs each year, ranging from 25.3% (28.2-22.8) to 55.8% (57.0-54.6). We also estimated that the total population declined by 12.58% (9.89-15.03) over the three-year period. This demonstrates that there is a high turnover of free-roaming dogs and that despite neutering a large number of dogs in an annual sterilisation campaign, the decline in population size was modest over a three-year time period. These findings have significant implications for the planning of rabies vaccination campaigns and population management programmes as well as highlighting the need for further research into the demographics of free-roaming, unowned dogs in India.

The Omicron, or Pango lineage B.1.1.529, variant of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) carries multiple spike mutations with high transmissibility and partial neutralizing antibody (nAb) escape. Vaccinated individuals show protection against severe disease, often attributed to primed cellular immunity. We investigated T and B cell immunity against B.1.1.529 in triple BioNTech BNT162b2 messenger RNA-vaccinated health care workers (HCWs) with different SARS-CoV-2 infection histories. B and T cell immunity against previous variants of concern was enhanced in triple-vaccinated individuals, but the magnitude of T and B cell responses against B.1.1.529 spike protein was reduced. Immune imprinting by infection with the earlier B.1.1.7 (Alpha) variant resulted in less durable binding antibody against B.1.1.529. Previously infection-naïve HCWs who became infected during the B.1.1.529 wave showed enhanced immunity against earlier variants but reduced nAb potency and T cell responses against B.1.1.529 itself. Previous Wuhan Hu-1 infection abrogated T cell recognition and any enhanced cross-reactive neutralizing immunity on infection with B.1.1.529.

INTRODUCTION B.1.1.529 (Omicron) and its subvariants pose new challenges for control of the COVID-19 pandemic. Although vaccinated populations are relatively protected from severe disease and death, countries with high vaccine uptake are experiencing substantial caseloads with breakthrough infection and frequent reinfection. RATIONALE We analyzed cross-protective immunity against B.1.1.529 (Omicron) in triple-vaccinated health care workers (HCWs) with different immune-imprinted histories of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection during the ancestral Wuhan Hu-1, B.1.1.7 (Alpha), and B.1.617.2 (Delta) waves and after infection during the B.1.1.529 (Omicron) wave in previously infection-naïve individuals and those with hybrid immunity, to investigate whether B.1.1.529 (Omicron) infection could further boost adaptive immunity. Spike subunit 1 (S1) receptor binding domain (RBD) and whole spike binding, live virus neutralizing antibody (nAb) potency, memory B cell (MBC) frequency, and T cell responses against peptide pools and naturally processed antigen were assessed. RESULTS B and T cell recognition and nAb potency were boosted against previous variants of concern (VOCs) in triple-vaccinated HCWs, but this enhanced immunity was attenuated against B.1.1.529 (Omicron) itself. Furthermore, immune imprinting after B.1.1.7 (Alpha) infection resulted in reduced durability of antibody binding against B.1.1.529 (Omicron), and S1 RBD and whole spike VOC binding correlated poorly with live virus nAb potency. Half of triple-vaccinated HCWs showed no T cell response to B.1.1.529 (Omicron) S1 processed antigen, and all showed reduced responses to the B.1.1.529 (Omicron) peptide pool, irrespective of SARS-CoV-2 infection history. Mapping T cell immunity in class II human leukocyte antigen transgenics showed that individual spike mutations could result in loss or gain of T cell epitope recognition, with changes to T cell effector and regulatory programs. Triple-vaccinated, previously infection-naïve individuals infected during the B.1.1.529 (Omicron) wave showed boosted cross-reactive S1 RBD and whole spike binding, live virus nAb potency, and T cell immunity against previous VOCs but less so against B.1.1.529 (Omicron) itself. Immune imprinting from prior Wuhan Hu-1 infection abrogated any enhanced cross-reactive antibody binding, T cell recognition, MBC frequency, or nAb potency after B.1.1.529 (Omicron) infection. CONCLUSION Vaccine boosting results in distinct, imprinted patterns of hybrid immunity with different combinations of SARS-CoV-2 infection and vaccination. Immune protection is boosted by B.1.1.529 (Omicron) infection in the triple-vaccinated, previously infection-naïve individuals, but this boosting is lost with prior Wuhan Hu-1 imprinting. This “hybrid immune damping” indicates substantial subversion of immune recognition and differential modulation through immune imprinting and may be the reason why the B.1.1.529 (Omicron) wave has been characterized by breakthrough infection and frequent reinfection with relatively preserved protection against severe disease in triple-vaccinated individuals.

This focus article has been prepared by Arthur Otter of the APHA Cattle Expert Group and Anna Brzozowska of the Surveillance Intelligence Unit.

Antibodies against SARS-CoV-2 are important to generate protective immunity, with convalescent plasma one of the first therapies approved. An alternative source of polyclonal antibodies suitable for upscaling would be more amendable to regulatory approval and widespread use. In this study, sheep were immunised with SARS-CoV-2 whole spike protein or one of the subunit proteins: S1 and S2. Once substantial antibody titres were generated, plasma was collected and samples pooled for each antigen. Non-specific antibodies were removed via affinity-purification to yield candidate products for testing in a hamster model of SARS-CoV-2 infection. Affinity-purified polyclonal antibodies to whole spike, S1 and S2 proteins were evaluated for in vitro for neutralising activity against SARS-CoV-2 Wuhan-like virus (Australia/VIC01/2020) and a recent variant of concern, B.1.1.529 BA.1 (Omicron), antibody-binding, complement fixation and phagocytosis assays were also performed. All antibody preparations demonstrated an effect against SARS-CoV-2 disease in the hamster model of challenge, with those raised against the S2 subunit providing the most promise. A rapid, cost-effective therapy for COVID-19 was developed which provides a source of highly active immunoglobulin specific to SARS-CoV-2 with multi-functional activity.

This study investigates possible effects of in utero exposure of rats to a low dose (125 mg/kg bw/day) and a high dose (750 mg/kg bw/day) of Diisononyl phthalate (DINP) during the masculinisation programming window (MPW) which is embryonic days 15.5-18.5 (e15.5 - e18.5). Dibutyl phthalate (DBP) was used at a high dose level (750 mg/kg bw/day) as an established positive control substance for anti-androgenic effects on the developing male reproductive tract. We focussed on the MPW and measured a multitude of biological endpoints at various life stages and applied state of the art histopathology staining techniques to refine the characterization of potential changes to the testis, beyond what is currently available with DINP. If DINP can mediate testicular dysgenesis (TDS) disorders, this exposure window would be sufficient to induce androgen impacts and alter male reproductive tract development as shown earlier in this validated experimental model with DBP. Overall, the results of this systematic comparison provide convincing evidence on the differences between the effects occurring with DBP and DINP. In contrast to what was seen with DBP, DINP did not cause cryptorchidism or hypospadias, had no effect on anogenital distance/anogenital index (AGD/AGi) and Leydig cell aggregates on e17.5 and e21.5 did not increase. With DINP no reduction of intratesticular testosterone, no effects on sperm motility and sperm count and no effect on adult testosterone or luteinizing hormone (LH) levels were seen. Our results demonstrate that DINP does not cause the adverse reproductive effects known to occur with DBP, a well-established endocrine disruptor.

The teaching of animal welfare in Indian veterinary education is limited. Current knowledge and attitudes to animal welfare and euthanasia, and the effect of a targeted educational intervention, were assessed in 84 Indian national and 49 non-Indian veterinarians attending a 2-week training course run by the Worldwide Veterinary Service in Tamil Nadu. A pre-intervention questionnaire, comprising knowledge and attitude questions on animal welfare and ethical issues, was completed. Fifteen students were then retained as a control group. The intervention group was exposed to a predesigned lecture and case studies (day 6). At the end of the course (day 12), another identical questionnaire was completed. Initially, there was no difference in knowledge of the control or intervention groups of Indian participants. Overall knowledge scores were lower in Indian participants compared with non-Indian participants ( p < 0.05). Both groups’ scores increased after the course ( p < 0.05), with the Indian participants improving the most. Indian participants’ attitudes were supportive of animal welfare and euthanasia prior to the intervention. Improvements in scores, with some reaching significance ( p < 0.05), were observed post-intervention. Non-Indian participants’ attitudes were more supportive of animal welfare and euthanasia with strongly agree/ strongly disagree chosen more frequently than Indian responses. Both groups’ self-assessment of their understanding of these topics improved post-intervention ( p < 0.01). No prominent differences were found in questionnaire responses in the control cohort. This study shows that a targeted educational intervention impacts on Indian veterinarians’ knowledge and attitudes toward animal welfare and euthanasia, and is relevant to organizations aiming to improve animal welfare standards in India.

IntroductionPulmonary hypertension is a major determinant of postnatal survival in infants with a congenital diaphragmatic hernia (CDH). The current care during the perinatal stabilisation period in these infants might contribute to the development of pulmonary hypertension after birth—in particular umbilical cord clamping before lung aeration. An ovine model of diaphragmatic hernia demonstrated that cord clamping after lung aeration, called physiological-based cord clamping (PBCC), avoided the initial high pressures in the lung vasculature while maintaining adequate blood flow, thereby avoiding vascular remodelling and aggravation of pulmonary hypertension. We aim to investigate if the implementation of PBCC in the perinatal stabilisation period of infants born with a CDH could reduce the incidence of pulmonary hypertension in the first 24 hours after birth.Methods and analysisWe will perform a multicentre, randomised controlled trial in infants with an isolated left-sided CDH, born at ≥35.0 weeks. Before birth, infants will be randomised to either PBCC or immediate cord clamping, stratified by treatment centre and severity of pulmonary hypoplasia on antenatal ultrasound. PBCC will be performed using a purpose-built resuscitation trolley. Cord clamping will be performed when the infant is considered respiratory stable, defined as a heart rate >100 bpm, preductal oxygen saturation >85%, while using a fraction of inspired oxygen of Ethics and disseminationCentral ethical approval was obtained from the Medical Ethical Committee of the Erasmus MC, Rotterdam, The Netherlands (METC 2019-0414). Local ethical approval will be obtained by submitting the protocol to the regulatory bodies and local institutional review boards.Trial registration numberNCT04373902.

Following findings in Northern America of SARS-CoV-2 infections in white-tailed deer, there is concern of similar infections in European deer and their potential as reservoirs of SARS-CoV-2 including opportunities for the emergence of new variants. UK deer sera were collected in 2020-2021 from 6 species and a hybrid with 1748 tested using anti-spike and anti-nucleocapsid serology assays. No samples were positive on both assays nor by surrogate neutralization testing. There is no evidence that spill-over infections of SARS-CoV-2 occurred from the human population to UK deer or that SARS-CoV-2 has been circulating in UK deer (over the study period). Although it cannot be ruled out, study results indicate that spill-over infections followed by circulation of SARS-CoV-2 to the most common European deer species is small.

Dog-mediated rabies kills tens of thousands of people each year in India, representing one third of the estimated global rabies burden. Whilst the World Health Organization (WHO), World Organization for Animal Health (OIE) and the Food and Agriculture Organization of the United Nations (FAO) have set a target for global dog-mediated human rabies elimination by 2030, examples of large-scale dog vaccination programs demonstrating elimination remain limited in Africa and Asia. We describe the development of a data-driven rabies elimination program from 2013 to 2019 in Goa State, India, culminating in human rabies elimination and a 92% reduction in monthly canine rabies cases. Smartphone technology enabled systematic spatial direction of remote teams to vaccinate over 95,000 dogs at 70% vaccination coverage, and rabies education teams to reach 150,000 children annually. An estimated 2249 disability-adjusted life years (DALYs) were averted over the program period at 526 USD per DALY, making the intervention ‘very cost-effective’ by WHO definitions. This One Health program demonstrates that human rabies elimination is achievable at the state level in India.

Aquatic ecosystems around the world are contaminated with a wide range of anthropogenic chemicals, including metals and organic pollutants, that originate from point and nonpoint sources. Many of these chemical contaminants have complex environmental cycles, are persistent and bioavailable, can be incorporated into aquatic food webs, and pose a threat to the health of wildlife and humans. Identifying appropriate sentinels that reflect bioavailability is critical to assessing and managing aquatic ecosystems impacted by contaminants. The objective of the present study is to review research on riparian spiders as sentinels of persistent and bioavailable chemical contaminants in aquatic ecosystems. Our review of the literature on riparian spiders as sentinels suggests that significant progress has been made during the last two decades of research. We identified 55 published studies conducted around the world in which riparian spiders (primarily of the families Tetragnathidae, Araneidae, Lycosidae, and Pisauridae) were used as sentinels of chemical contamination of lotic, lentic, and estuarine systems. For several contaminants, such as polychlorinated biphenyls (PCBs), Hg, and Se, it is now clear that riparian spiders are appropriate sentinels. However, many contaminants and factors that could impact chemical concentrations in riparian spiders have not been well characterized. Further study of riparian spiders and their potential role as sentinels is critical because it would allow for development of national-scale programs that utilize riparian spiders as sentinels to monitor chemical contaminants in aquatic ecosystems. A riparian spider sentinel program in the United States would be complementary to existing national sentinel programs, including those for fish and immature dragonflies. Environ Toxicol Chem 2022;41:499-514. © 2021 SETAC.

BackgroundFew studies have quantified aerosol concentrations of SARS-CoV-2 in hospitals and long-term care homes, and fewer still have examined samples for viability. This information is needed to clarify transmission risks beyond close contact.MethodsWe deployed particulate air samplers in rooms with COVID-19 positive patients in hospital ward and ICU rooms, rooms in long-term care homes experiencing outbreaks, and a correctional facility experiencing an outbreak. Samplers were placed between 2 and 3 meters from the patient. Aerosol (small liquid particles suspended in air) samples were collected onto gelatin filters by Ultrasonic Personal Air Samplers (UPAS) fitted with 3), and with a Coriolis Biosampler over 10 minutes (total 1.5m3). Samples were assayed for viable SARS-CoV-2 virus and for the viral genome by multiplex PCR using the E and N protein target sequences. We validated the sampling methods by inoculating gelatin filters with viable vesicular stomatitis virus (VSV), and with three concentrations of viable SARS-CoV-2, operating personal samplers for 16hrs, and quantifying viable virus recovery by TCID50 assay.ResultsIn total, 138 samples were collected from 99 rooms. RNA samples were positive in 9.1% (6/66) of samples obtained with the UPAS 2.5µm samplers, 13.5% (7/52) with the UPAS 10µm samplers, and 10.0% (2/20) samples obtained with the Coriolis samplers. Culturable virus was not recovered in any samples. Viral RNA was detected in 10.9% of the rooms sampled. There was no significant difference in viral RNA recovery between the different room locations or samplers. Method development experiments indicated minimal loss of SARS-CoV-2 viability via the personal air sampler operation.Key FindingsAlthough a subset of aerosol samples exhibited detectable SARS-CoV-2 RNA at low titres, the presence of viable SARS-CoV-2 virus in aerosols appears to be infrequent at >2m distance.

INTRODUCTION: Studies of acute hypersensitivity reactions in pediatric populations receiving Crotalidae Polyvalent Immune Fab (CPIF) are complicated by small size, wide age ranges, and diverse definitions of such reactions. METHODS: This is a retrospective chart review of patients aged 13 years or younger treated with CPIF for Crotalid envenomation from November 2006 to 2016. The primary outcome was the presence of an acute hypersensitivity reaction to CPIF and was defined as the development of any of the following symptoms within 3 hours of initiation of CPIF infusion: urticaria, wheezing or respiratory distress, angioedema, hypotension, nausea, and/or vomiting. Demographics, CPIF dose to control and total dose, bite location, level of care, and length of stay were also recorded. RESULTS: Thirty-four patients were ultimately treated with CPIF. Ages ranged from 10 months to 13 years. Twenty-one patients (60%) were male, 24 (70.6%) were admitted to the ICU, and the median length of stay was 2 days with a range of 1–11 days. Zero patients developed an acute hypersensitivity reaction to CPIF. CONCLUSION: Acute hypersensitivity reactions to CPIF did not occur in this cohort. Such reactions are rare with the use of CPIF in pediatric patients.

Millipedes are among the most important processors of leaf litter in temperate forests. Through consumption of leaf litter, millipedes may be exposed to mercury that accumulates in leaf tissues prior to senescence. To investigate mercury uptake in millipedes, Narceus spp. were collected from a remote site in the southern Appalachian Mountains, an area known to receive high mercury deposition. Additionally, aquatic primary consumers (larval caddisflies and stoneflies), brook trout (Salvelinus fontinalis) and rainbow trout (Oncorhynchus mykiss) were collected from the same site for comparisons of mercury concentrations and percent methylmercury. Bioaccumulation factors for millipedes were 18.5 and 20.2 for total and methylmercury, respectively. At this site, the mean THg concentration in millipedes was ~ 10 × greater than both brook trout and rainbow trout and ~ 200 × greater than that of aquatic primary consumers. Millipede THg concentrations ranged from 222 to 1620 ng/g ww in an area where EPA fish consumption criteria (300 ng/g MeHg in fish tissue, ww) were not exceeded. The mean percent methylmercury in millipedes was 1.4%, suggesting these animals were accumulating large quantities of inorganic mercury.

Flexible, rapid, and predictive approaches that do not require the use of large numbers of vertebrate test animals are needed because the chemical universe remains largely untested for potential hazards. Development of robust new approach methodologies and nontesting approaches requires the use of existing information via curated, integrated data sets. The ecological threshold of toxicological concern (ecoTTC) represents one such new approach methodology that can predict a conservative de minimis toxicity value for chemicals with little or no information available. For the creation of an ecoTTC tool, a large, diverse environmental data set was developed from multiple sources, with harmonization, characterization, and information quality assessment steps to ensure that the information could be effectively organized and mined. The resulting EnviroTox database contains 91 217 aquatic toxicity records representing 1563 species and 4016 unique Chemical Abstracts Service numbers and is a robust, curated database containing high‐quality aquatic toxicity studies that are traceable to the original information source. Chemical‐specific information is also linked to each record and includes physico‐chemical information, chemical descriptors, and mode of action classifications. Toxicity data are associated with the physico‐chemical data, mode of action classifications, and curated taxonomic information for the organisms tested. The EnviroTox platform also includes 3 analysis tools: a predicted‐no‐effect concentration calculator, an ecoTTC distribution tool, and a chemical toxicity distribution tool. Although the EnviroTox database and tools were originally developed to support ecoTTC analysis and development, they have broader applicability to the field of ecological risk assessment. Environ Toxicol Chem 2019;9999:1–12. © 2019 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.

Host-associated microbial communities play important roles in wildlife health, but these dynamics can be influenced by environmental factors. Urbanization has numerous effects on wildlife; however, the degree to which wildlife-associated bacterial communities and potential bacterial pathogens vary across urban-rural/native habitat gradients remains largely unknown. We used

Rabies is a fatal zoonotic disease transmitted by the bite of a rabid animal. More than 95% of the human rabies cases in India are attributed to exposure to rabid dogs. This study evaluated the utility of a lateral flow immunochromatographic assay (LFA) (Anigen Rapid Rabies Ag Test Kit, Bionote, Hwaseong-si, Korea) for rapid post mortem diagnosis of rabies in dogs. Brain tissue was collected from 202 animals that were screened through the Government of Goa rabies surveillance system. The brain tissue samples were obtained from 188 dogs, nine cats, three bovines, one jackal and one monkey. In addition, 10 dogs that died due to trauma from road accidents were included as negative controls for the study. The diagnostic performance of LFA was evaluated using results from direct fluorescence antibody test (dFT), the current gold standard post mortem test for rabies infection. Three samples were removed from the analysis as they were autolysed and not fit for testing by dFT. Of the 209 samples tested, 117 tested positive by LFA and 92 tested negative, while 121 tested positive by dFT and 88 tested negative. Estimates of LFA sensitivity and specificity were 0.96 (95% CI 0.91&ndash, 0.99) and 0.99 (95% CI 0.94&ndash, 1.00), respectively. The LFA is a simple and low-cost assay that aids in the rapid diagnosis of rabies in the field without the need for expensive laboratory equipment or technical expertise. This study found that Bionote LFA has potential as a screening tool in rabies endemic countries.

Doxorubicin is a commonly used anticancer agent that can cause debilitating and irreversible cardiac injury. The initiating mechanisms contributing to this side effect remain unknown, and current preventative strategies offer only modest protection. Using stem-cell–derived cardiomyocytes from patients receiving doxorubicin, we probed the transcriptomic landscape of solute carriers and identified organic cation transporter 3 (OCT3) (SLC22A3) as a critical transporter regulating the cardiac accumulation of doxorubicin. Functional validation studies in heterologous overexpression models confirmed that doxorubicin is transported into cardiomyocytes by OCT3 and that deficiency of OCT3 protected mice from acute and chronic doxorubicin-related changes in cardiovascular function and genetic pathways associated with cardiac damage. To provide proof-of-principle and demonstrate translational relevance of this transport mechanism, we identified several pharmacological inhibitors of OCT3, including nilotinib, and found that pharmacological targeting of OCT3 can also preserve cardiovascular function following treatment with doxorubicin without affecting its plasma levels or antitumor effects in multiple models of leukemia and breast cancer. Finally, we identified a previously unrecognized, OCT3-dependent pathway of doxorubicin-induced cardiotoxicity that results in a downstream signaling cascade involving the calcium-binding proteins S100A8 and S100A9. These collective findings not only shed light on the etiology of doxorubicin-induced cardiotoxicity, but also are of potential translational relevance and provide a rationale for the implementation of a targeted intervention strategy to prevent this debilitating side effect.

Purpose Ventilation-induced tumour motion remains a challenge for the accuracy of proton therapy treatments in lung patients. We investigated the feasibility of using a 4D virtual CT (4D-vCT) approach based on deformable image registration (DIR) and motion-aware 4D CBCT reconstruction (MA-ROOSTER) to enable accurate daily proton dose calculation using a gantry-mounted CBCT scanner tailored to proton therapy. Methods Ventilation correlated data of 10 breathing phases were acquired from a porcine ex-vivo functional lung phantom using CT and CBCT. 4D-vCTs were generated by (1) DIR of the mid-position 4D-CT to the mid-position 4D-CBCT (reconstructed with the MA-ROOSTER) using a diffeomorphic Morphons algorithm and (2) subsequent propagation of the obtained mid-position vCT to the individual 4D-CBCT phases. Proton therapy treatment planning was performed to evaluate dose calculation accuracy of the 4D-vCTs. A robust treatment plan delivering a nominal dose of 60 Gy was generated on the average intensity image of the 4D-CT for an approximated internal target volume (ITV). Dose distributions were then recalculated on individual phases of the 4D-CT and the 4D-vCT based on the optimized plan. Dose accumulation was performed for 4D-vCT and 4D-CT using DIR of each phase to the mid position, which was chosen as reference. Dose based on the 4D-vCT was then evaluated against the dose calculated on 4D-CT both, phase-by-phase as well as accumulated, by comparing dose volume histogram (DVH) values (Dmean, D2%, D98%, D95%) for the ITV, and by a 3D-gamma index analysis (global, 3%/3 mm, 5 Gy, 20 Gy and 30 Gy dose thresholds). Results Good agreement was found between the 4D-CT and 4D-vCT-based ITV-DVH curves. The relative differences ((CT-vCT)/CT) between accumulated values of ITV Dmean, D2%, D95% and D98% for the 4D-CT and 4D-vCT-based dose distributions were −0.2%, 0.0%, −0.1% and −0.1%, respectively. Phase specific values varied between −0.5% and 0.2%, −0.2% and 0.5%, −3.5% and 1.5%, and −5.7% and 2.3%. The relative difference of accumulated Dmean over the lungs was 2.3% and Dmean for the phases varied between −5.4% and 5.8%. The gamma pass-rates with 5 Gy, 20 Gy and 30 Gy thresholds for the accumulated doses were 96.7%, 99.6% and 99.9%, respectively. Phase-by-phase comparison yielded pass-rates between 86% and 97%, 88% and 98%, and 94% and 100%. Conclusions Feasibility of the suggested 4D-vCT workflow using proton therapy specific imaging equipment was shown. Results indicate the potential of the method to be applied for daily 4D proton dose estimation.

Annual peaks in reproductive activity have been identified in multiple domestic dog populations. However, there is little evidence to describe how these peaks may be associated with environmental factors such as daylength, which plays a well-established role in breeding patterns of seasonally-reproductive species. Data were collected 2016-2020 during 7,743 and 4,681 neuter surgeries on adult female unowned free-roaming dogs in veterinary clinics in Goa and Tamil Nadu respectively. Temperature, precipitation, relative humidity, and daylength data were gathered for time periods preceding the neuter surgery that may have influenced the likelihood of pregnancy (potential influence periods). A multivariable generalised additive model was used to assess the relationship between these factors and pregnancy. The prevalence of pregnancy varied by month in both locations indicating seasonality of reproduction in these groups. The annual pattern was more distinct in Goa with a peak in pregnancies between September and December. In Goa, decreasing daylength was associated with a higher probability of pregnancy (p = 0.040). Decreasing temperature was associated with decreasing probability of pregnancy in the Nilgiris (p = 0.034). Bitches had a median of 6 foetuses, with no evidence of seasonal variation. Environmental factors were associated with patterns of pregnancy in free-roaming dogs, however statistically-significant factors varied by geographical location. Establishing local seasonal patterns of breeding in free-roaming dogs and assessing their relationship with environmental influences is recommended to facilitate effective and efficient population management strategies, which aim to reduce conflict between human and free-roaming dog populations.

There is growing concern over the health of North American birds, with evidence suggesting substantial population declines. Spiders are prominent dietary items for many bird species and mediate the transfer of contaminants to arachnivorous birds that consume them. Few studies have investigated the potential risk the spider exposure pathway poses to these birds because most studies have focused on piscivores. In the present study, we developed new chronic and acute As, Cd, Cu, Pb, Ni, Se, Zn, and MeHg spider-based avian wildlife values (SBAWVs) for multiple adult and nestling birds (primarily passerines) and then used the newly generated SBAWVs to characterize the risk to birds across 2 study areas: 1) 5 reaches in the southern Appalachian Mountains, an area with substantial mercury deposition but minimal anthropogenic impact, and 2) 4 reaches adjacent to the Emory River, an area impacted by the largest fly coal-ash spill in US history. We identified MeHg and Cu, Pb, Se, and Zn as contaminants of potential concern (COPC) at the Appalachian Mountain and Emory River study areas, respectively, based on dietary exposure of aquatic contaminants via riparian spiders. The identification of COPC at both study areas due to dietary spider exposure is notable not only because the spider exposure pathway has largely been uninvestigated at these sites but also because the aquatic systems in both areas have been studied extensively. Significant differences in MeHg concentrations were detected among spider taxa and suggest that the selection of spider taxa can impact risk characterization. These results indicate that the spider exposure pathway is important to consider when assessing potential risk, particularly for passerine birds. Environ Toxicol Chem 2020;39:2314-2324. © 2020 SETAC.

Inhibitors that block the programmed cell death-1 (PD-1) pathway can potentiate endogenous antitumor immunity and have markedly improved cancer survival rates across a broad range of indications. However, these treatments work for only a minority of patients. The efficacy of anti-PD-1 inhibitors may be extended by cytokines, however, the incorporation of cytokines into therapeutic regimens has significant challenges. In their natural form when administered as recombinant proteins, cytokine treatments are often associated with low response rates. Most cytokines have a short half-life which limits their exposure and efficacy. In addition, cytokines can activate counterregulatory pathways, in the case of immune-potentiating cytokines this can lead to immune suppression and thereby diminish their potential efficacy. Improving the drug-like properties of natural cytokines using protein engineering can yield synthetic cytokines with improved bioavailability and tissue targeting, allowing for enhanced efficacy and reduced off-target effects. Using structure guided engineering we have designed a novel class of antibody-cytokine fusion proteins consisting of a PD-1 targeting antibody fused together with an interleukin-21 (IL-21) cytokine mutein. Our bifunctional fusion proteins can block PD-1/programmed death-ligand 1 (PD-L1) interaction whilst simultaneously delivering IL-21 cytokine to PD-1 expressing T cells. Targeted delivery of IL-21 can improve T cell function in a manner that is superior to anti-PD-1 monotherapy. Fusion of engineered IL-21 variants to anti-PD1 antibodies can improve the drug-like properties of IL-21 cytokine leading to improved cytokine serum half-life allowing for less frequent dosing. In addition, we show that targeted delivery of IL-21 can minimize any potential detrimental effect on local antigen-presenting cells. A highly attenuated IL-21 mutein variant (R9E:R76A) fused to a PD-1 antibody provides protection in a humanized mouse model of cancer that is refractory to anti-PD-1 monotherapy. Collectively, our preclinical data demonstrate that this approach may improve upon and extend the utility of anti-PD-1 therapeutics currently in the clinic.

Human exposure to persistent, nonbiological nanoparticles and microparticles via the oral route is continuous and large scale (1012 -1013 particles per day per adult in Europe). Whether this matters or not is unknown but confirmed health risks with airborne particle exposure warns against complacency. Murine models of oral exposure will help to identify risk but, to date, lack validation or relevance to humans. This work addresses that gap. It reports i) on a murine diet, modified with differing concentrations of the common dietary particle, food grade titanium dioxide (fgTiO2 ), an additive of polydisperse form that contains micro- and nano-particles, ii) that these diets deliver particles to basal cells of intestinal lymphoid follicles, exactly as is reported as a "normal occurrence" in humans, iii) that confocal reflectance microscopy is the method of analytical choice to determine this, and iv) that food intake, weight gain, and Peyer's patch immune cell profiles, up to 18 weeks of feeding, do not differ between fgTiO2 -fed groups or controls. These findings afford a human-relevant and validated oral dosing protocol for fgTiO2 risk assessment as well as provide a generalized platform for application to oral exposure studies with nano- and micro-particles.

Hypotheses on regional song variation ("dialects") assume that dialects remain stable within regions, are distinct between regions, and persist within populations over extensive periods [1-3]. Theories to explain dialects focus on mechanisms that promote persistence of regional song variants despite gene flow between regions [4-6], such as juveniles settling in non-natal populations retaining only those songs from their repertoires that match neighbors [7, 8]. It would be considered atypical for a novel song variant to invade and replace the established regional variant. Yet some studies have reported song variants shifting rapidly over time within populations [9-11]. White-throated sparrows, Zonotrichia albicolis, for example, traditionally sing a whistled song terminating in a repeated triplet of notes [12], which was the ubiquitous variant in surveys across Canada in the 1960s [13]. However, doublet-ending songs emerged and replaced triplet-ending songs west of the Rocky Mountains sometime between 1960 and 2000 [11] and appeared just east of the Rockies in the 2000s [14]. From recordings collected over two decades across North America, we show that doublet-ending song has now spread at a continental scale. Using geolocator tracking, we confirm that birds from western Canada, where doublet-ending songs originated, overwinter with birds from central Canada, where the song initially spread. This suggests a potential mechanism for spread through song tutoring on wintering grounds. Where the new song variant has spread, it rose from a rare variant to the sole, regional song type, as predicted by the indirect biased transmission hypothesis [10]. VIDEO ABSTRACT.

Doxorubicin is a frequently used anticancer drug to treat many types of tumors, such as breast cancer or bronchial carcinoma. The clinical use of doxorubicin is limited by its poorly predictable cardiotoxicity, the reasons of which are so far not fully understood. The drug is a substrate of several efflux transporters such as P-gp or BCRP and was recently reported to be a substrate of cation uptake transporters. To evaluate the potential role of transporter proteins in the accumulation of doxorubicin at its site of action (e.g., mammary carcinoma cells) or adverse effects (e.g., heart muscle cells), we studied the expression of important uptake and efflux transporters in human breast cancer and cardiac tissue, and investigated the affinity of doxorubicin to the identified transporters. The cellular uptake studies on doxorubicin were performed with OATP1A2*1, OATP1A2*2, and OATP1A2*3-overexpressing HEK293 cells, as well as OCT1-, OCT2-, and OCT3- overexpressing MDCKII cells. To assess the contribution of transporters to the cytotoxic effect of doxorubicin, we determined the cell viability in the presence and absence of transporter inhibitors in different cell lines. Several transporters, including P-gp, BCRP, OCT1, OCT3, and OATP1A2 were expressed in human heart and/or breast cancer tissue. Doxorubicin could be identified as a substrate of OCT1, OCT2, OCT3, and OATP1A2. The cellular uptake into cells expressing genetic OATP1A2 variants was markedly reduced and correlated well with the increased cellular viability. Inhibition of OATP1A2 (naringin) and OCT transporters (1-methyl-4-phenylpyridinium) resulted in a significant decrease of doxorubicin-mediated cytotoxicity in cell lines expressing the respective transporters. Similarly, the excipient Cremophor EL significantly inhibited the OCT1-3- and OATP1A2-mediated cellular uptake and attenuated the cytotoxicity of doxorubicin. In conclusion, genetic and environmental-related variability in the expression and function of these transporters may contribute to the substantial variability seen in terms of doxorubicin efficacy and toxicity.

This focus article has been prepared by Arthur Otter of the APHA Cattle Expert Group.

Dextromethorphan (DXM) acts as cough suppressant via its central action. Cell-protective effects of this drug have been reported in peripheral tissues, making DXM potentially useful for treatment of several common human diseases, such as type 2 diabetes mellitus (T2DM). Pancreatic islets are among the peripheral tissues that positively respond to DXM, and anti-diabetic effects of DXM were observed in two placebo-controlled, randomized clinical trials in humans with T2DM. Since these effects were associated with central side effects, we here developed chemical derivatives of DXM that pass the blood-brain barrier to a significantly lower extent than the original drug. We show that basic nitrogen-containing residues block central adverse events of DXM without reducing its anti-diabetic effects, including the protection of human pancreatic islets from cell death. These results show how to chemically modify DXM, and possibly other morphinans, as to exclude central side effects, while targeting peripheral tissues, such as pancreatic islets.

Understanding the chemistry of milk and its components is critical to the production of consistent, high-quality dairy products as well as the development of new dairy ingredients. Over the past 100 yr we have gone from believing that milk has only 3 protein fractions to identifying all the major and minor types of milk proteins as well as discovering that they have genetic variants. The structure and physical properties of most of the milk proteins have been extensively studied. The structure of the casein micelle has been the subject of many studies, and the initial views on submicelles have given way to the current model of the micelle as being assembled as a result of the concerted action of several types of interactions (including hydrophobic and the formation of calcium phosphate nanoclusters). The benefits of this improved knowledge of the type and nature of casein interactions include better control of the cheesemaking process, more functional milk powders, development of new products such as cream liqueurs, and expanded food applications. Increasing knowledge of proteins and minerals was paralleled by developments in the analysis of milk fat and its synthesis together with greater knowledge of its packaging in the milk fat globule membrane. Advances in analytical techniques have been essential to the isolation and characterization of milk components. Milk testing has progressed from gross compositional analyses of the fat and total solids content to the rapid analysis of milk for a wide range of components for various purposes, such as diagnostic issues related to animal health. Up to the 1950s, research on dairy chemistry was mostly focused on topics such as protein fractionation, heat stability, acid-base buffering, freezing point, and the nature of the calcium phosphate present in milk. Between the 1950s and 1970s, there was a major focus on identifying all the main protein types, their sequences, variants, association behavior, and other physical properties. During the 1970s and 1980s, one of the major emphases in dairy research was on protein functionality and fractionation processes. The negative cloud over dairy fat has lifted recently due to multiple reviews and meta-analyses showing no association with chronic issues such as cardiovascular disease, but changing consumer misconceptions will take time. More recently, there has been a great deal of interest in the biological and nutritional components in milk and how these materials were uniquely designed by the cow to achieve this type of purpose.

Background Pigment-grade titanium dioxide (TiO2) particles are an additive to some foods (E171 on ingredients lists), toothpastes, and pharma−/nutraceuticals and are absorbed, to some extent, in the human intestinal tract. TiO2 can act as a modest adjuvant in the secretion of the pro-inflammatory cytokine interleukin 1β (IL-1β) when triggered by common intestinal bacterial fragments, such as lipopolysaccharide (LPS) and/or peptidoglycan. Given the variance in human genotypes, which includes variance in genes related to IL-1β secretion, we investigated whether TiO2 particles might, in fact, be more potent pro-inflammatory adjuvants in cells that are genetically susceptible to IL-1β-related inflammation. Methods We studied bone marrow-derived macrophages from mice with a mutation in the nucleotide-binding oligomerisation domain-containing 2 gene (Nod2 m/m), which exhibit heightened secretion of IL-1β in response to the peptidoglycan fragment muramyl dipeptide (MDP). To ensure relevance to human exposure, TiO2 was food-grade anatase (119 ± 45 nm mean diameter ± standard deviation). We used a short ‘pulse and chase’ format: pulsing with LPS and chasing with TiO2 +/− MDP or peptidoglycan. Results IL-1β secretion was not stimulated in LPS-pulsed bone marrow-derived macrophages, or by chasing with MDP, and only very modestly so by chasing with peptidoglycan. In all cases, however, IL-1β secretion was augmented by chasing with TiO2 in a dose-dependent fashion (5–100 μg/mL). When co-administered with MDP or peptidoglycan, IL-1β secretion was further enhanced for the Nod2 m/m genotype. Tumour necrosis factor α was triggered by LPS priming, and more so for the Nod2 m/m genotype. This was enhanced by chasing with TiO2, MDP, or peptidoglycan, but there was no additive effect between the bacterial fragments and TiO2. Conclusion Here, the doses of TiO2 that augmented bacterial fragment-induced IL-1β secretion were relatively high. In vivo, however, selected intestinal cells appear to be loaded with TiO2, so such high concentrations may be ‘exposure-relevant’ for localised regions of the intestine where both TiO2 and bacterial fragment uptake occurs. Moreover, this effect is enhanced in cells from Nod2 m/m mice indicating that genotype can dictate inflammatory signalling in response to (nano)particle exposure. In vivo studies are now merited.

Tauopathies include a variety of neurodegenerative diseases associated with the pathological aggregation of hyperphosphorylated tau, resulting in progressive cognitive decline and motor impairment. The underlying mechanism for motor deficits related to tauopathy is not yet fully understood. Here, we use a novel transgenic tau mouse line, Tau 58/4, with enhanced neuron-specific expression of P301S mutant tau to investigate the motor abnormalities in association with the peripheral nervous system. Using stationary beam, gait, and rotarod tests, motor deficits were found in Tau 58/4 mice already 3 months after birth, which deteriorated during aging. Hyperphosphorylated tau was detected in the cell bodies and axons of motor neurons. At the age of 9 and 12 months, significant denervation of the neuromuscular junction in the extensor digitorum longus muscle was observed in Tau 58/4 mice, compared to wild-type mice. Muscle hypotrophy was observed in Tau 58/4 mice at 9 and 12 months. Using electron microscopy, we observed ultrastructural changes in the sciatic nerve of 12-month-old Tau 58/4 mice indicative of the loss of large axonal fibers and hypomyelination (assessed by g-ratio). We conclude that the accumulated hyperphosphorylated tau in the axon terminals may induce dying-back axonal degeneration, myelin abnormalities, neuromuscular junction denervation, and muscular atrophy, which may be the mechanisms responsible for the deterioration of the motor function in Tau 58/4 mice. Tau 58/4 mice represent an interesting neuromuscular degeneration model, and the pathological mechanisms might be responsible for motor signs observed in some human tauopathies.

Purpose. We previously identified an association between CC22 meticillin-resistant Staphylococcus aureus (MRSA) bloodstream infection isolates with an elevated vancomycin MIC (V-MIC) in the susceptible range (1.5–2 mg l−1) and endocarditis. This study explores whether these isolates have a specific phenotype consistent with the clinical findings. Methodology. CC22 and CC30 MRSA isolates with high (1.5–2 mg l−1) and low (≤0.5 mg l−1) V-MICs were tested for fibrinogen and fibronectin binding, virulence in a Galleria mellonella caterpillar model, phenol soluble modulin production and accessory gene regulator (agr) expression. Results. CC22 high V-MIC, but not CC30 high V-MIC isolates, showed sustained fibrinogen binding through a stationary growth phase and increased PSM production, specifically PSMα1, compared with respective low V-MIC isolates. Expression was lower in both CC22 and CC30 high V-MIC isolates compared with respective low V-MIC isolates, although there was no associated reduction in virulence in the caterpillar model. Conclusions. The identification of a distinct phenotype for CC22 high V-MIC isolates supports the hypothesis that bacterial factors contribute to the mechanism underlying their association with endocarditis. Further study of these isolates could shed light on the molecular mechanism of endocarditis in humans.

A physiologically based pharmacokinetic (PBPK) model for di-isononyl phthalate (DiNP) was developed by adapting the existing models for di(2-ethylhexyl) phthalate (DEHP) and di-butylphthalate (DBP). Both pregnant rat and human time-course plasma and urine data were used to address the hydrolysis of DiNP in intestinal tract, plasma, and liver as well as hepatic oxidative metabolism and conjugation of the monoester and primary oxidative metabolites. Data in both rats and humans were available to inform the uptake and disposition of mono-isononyl phthalate (MiNP) as well as the three primary oxidative metabolites including hydroxy (7-OH)-, oxo (7-OXO)-, and carboxy (7-COX)-monoisononyl phthalate in plasma and urine. The DiNP model was reliable over a wide range of exposure levels in the pregnant rat as well as the two low exposure levels in humans including capturing the nonlinear behavior in the pregnant rat after repeated 750 mg/kg/day dosing. The presented DiNP PBPK model in pregnant rat and human, based upon an extensive kinetic dataset in both species, may provide a basis for assessing human equivalent exposures based upon either rodent or in vitro points of departure.

Human exposure to persistent, nonbiological nanoparticles and microparticles via the oral route is continuous and large scale (1012 -1013 particles per day per adult in Europe). Whether this matters or not is unknown but confirmed health risks with airborne particle exposure warns against complacency. Murine models of oral exposure will help to identify risk but, to date, lack validation or relevance to humans. This work addresses that gap. It reports i) on a murine diet, modified with differing concentrations of the common dietary particle, food grade titanium dioxide (fgTiO2 ), an additive of polydisperse form that contains micro- and nano-particles, ii) that these diets deliver particles to basal cells of intestinal lymphoid follicles, exactly as is reported as a "normal occurrence" in humans, iii) that confocal reflectance microscopy is the method of analytical choice to determine this, and iv) that food intake, weight gain, and Peyer's patch immune cell profiles, up to 18 weeks of feeding, do not differ between fgTiO2 -fed groups or controls. These findings afford a human-relevant and validated oral dosing protocol for fgTiO2 risk assessment as well as provide a generalized platform for application to oral exposure studies with nano- and micro-particles.

BACKGROUND: Preventable wounds are a common welfare issue in working donkeys in many countries. In the Bukombe District of Tanzania, there are estimated to be 3000 working donkeys, used primarily to transport loads for direct income generation. For historical reasons, oxen-yoke carts are used; their design is inappropriate for donkeys and results in serious neck wounds. The project aim was to assess the prevalence and nature of wounds in working donkeys.METHODS: In November 2018, 148 donkeys owned by 48 owners were examined, and data were collected.RESULTS: The study revealed that one or more wounds were present in 56.1% of the population and yoke-related, dorsal neck wounds comprised 79.5% of these. These wounds ranged in surface area from 1 cm2 to 300 cm2. Clinically, 96.6% of all wound types were superficial and the majority of these (51.1%) were granulating.CONCLUSION: These data will enable the future evaluation of targeted interventions aimed at reducing the prevalence of these specific wounds.

Cheilostome Bryozoa Anoteropora latirostris, a colonial marine invertebrate, constructs its skeleton from calcite and aragonite. This study presents firstly correlated multi-scale electron microscopy, micro-computed tomography, electron backscatter diffraction and NanoSIMS mapping. We show that all primary, coarse-grained platy calcitic lateral walls are covered by fine-grained fibrous aragonite. Vertical lateral walls separating autozooid chambers have aragonite only on their distal side. This type of asymmetric mineralization of lateral walls results from the vertical arrangement of the zooids at the growth margins of the colony and represents a type of biomineralization previously unknown in cheilostome bryozoans. NanoSIMS mapping across the aragonite-calcite interface indicates an organic layer between both mineral phases, likely representing an organic template for biomineralization of aragonite on the calcite layer. Analysis of crystallographic orientations show a moderately strong crystallographic preferred orientation (CPO) for calcite (7.4 times random orientation) and an overall weaker CPO for aragonite (2.4 times random orientation) with a high degree of twinning (45%) of the aragonite grains. The calculated Young’s modulus for the CPO map shows a weak mechanical direction perpendicular to the colony’s upper surface facilitating this organism’s strategy of clonal reproduction by fragmentation along the vertical zooid walls.

Derivation of Predicted No Effect Concentrations (PNECs) for aquatic systems is the primary deterministic form of hazard extrapolation used in environmental risk assessment. Depending on the data availability, different regulatory jurisdictions apply application factors (AFs) to the most sensitive measured endpoint to derive the PNEC for a chemical. To assess differences in estimated PNEC values, two PNEC determination methodologies were applied to a curated public database using the EnviroTox Platform ( www.EnviroToxdatabase.org ). PNECs were derived for 3647 compounds using derivation procedures based on example US EPA and a modified European Union chemical registration procedure to allow for comparisons. Ranked probability distributions of PNEC values were developed and 5th percentile values were calculated for the entire dataset and scenarios where full acute or full chronic data sets were available. The lowest PNEC values indicated categorization based on chemical attributes and modes of action would lead to improved extrapolations. Full acute or chronic datasets gave measurably higher 5th percentile PNEC values. Algae were under-represented in available ecotoxicity data but drove PNECs disproportionately. Including algal inhibition studies will be important in understanding chemical hazards. The PNEC derivation logic flows are embedded in the EnviroTox Platform providing transparent and consistent PNEC derivations and PNEC distribution calculations.

The susceptibility of Tennessee's Appalachian Mountains to anthropogenic stressors has remained largely uninvestigated likely due to a lack of known point source contamination. However, a growing body of scientific evidence suggests that depositional inputs can lead to concerning levels of contamination, even in remote areas. To investigate potential concerns, water quality parameters, contaminants in water (nitrogen, TSS, and metals), and contaminants in eastern brook trout (mercury, polychlorinated biphenyls [PCBs], organochlorine [OC] pesticides, dioxins, furans, and phthalates) were measured in four Appalachian Mountain streams from 2015 to 2017. Concentrations were compared to literature and/or model-derived (e.g., biotic ligand model) threshold values to determine whether levels exceeded those acceptable for stream health. Dioxins and furans were detectable in fish tissue at all sites with an average 2,3,7,8-tetrachlorodinbenzodioxin toxicity equivalence (TEQ) of 0.0015 ng/kg. Concentrations of PCBs, phthalates, and organochlorine pesticides were never above analytical quantitation limits, although several OC pesticides (e.g., alpha-chlordane) were detectable in fish. Aluminum concentrations in water were found at levels shown previously to cause mortality in brook trout during acidic rain events. The average whole-body methylmercury concentrations in fish among sites were 0.037 ± 0.003 μg/kg and were on average 75 ± 2% of total mercury.

The effective management of stray dogs is critically important in any rabies vaccination programme. In many rabies-endemic countries, stray dogs represent a significant proportion of both the free-roaming and total dog populations, and to ensure that rabies elimination programmes are successful, it is essential that this portion of the dog population (stray dogs) is vaccinated at high coverage. However, there are a number of challenges to managing and delivering rabies vaccinations to stray dogs. This paper will review the most pertinent issues relating to the problem of stray dogs and rabies elimination, focusing on the challenges relating to the estimation of population size, vaccination administration and vaccine coverage assessment in stray dog populations. The authors will highlight how these challenges can be overcome, notably by reviewing the census techniques that have been employed to assess stray dog populations, which then facilitate the planning and design of vaccination programmes. In addition, they discuss the range of vaccination strategies that are available and that have been used to vaccinate stray dogs and review how vaccination coverage can be assessed in this population. Finally, the benefits of population management in stray dogs through neutering programmes are reviewed. In short, this paper highlights the importance of ensuring that stray dogs, as well as owned dogs, are included in vaccination programmes and emphasises that many of the challenges related to protecting stray dogs from rabies can be overcome.L’efficacité de la gestion des populations de chiens errants est un aspect essentiel de tout programme de vaccination contre la rage. Dans de nombreux pays où la rage est endémique, les chiens errants constituent un segment important des populations de chiens laissés en liberté et plus généralement de la population canine totale, de sorte que le succès des programmes d’élimination de la rage repose sur l’impératif d’assurer une forte couverture vaccinale au sein de cette partie de la population canine. Toutefois, s’agissant des chiens errants la gestion de la vaccination et l’administration des vaccins antirabiques présentent des difficultés particulières. Les auteurs font le point sur les principales questions posées par la problématique des chiens errants dans une perspective d’élimination de la rage, qui concernent notamment la difficulté d’évaluer la taille des populations concernées, d’administrer les vaccins dans ce cadre et d’évaluer la couverture vaccinale obtenue. Les auteurs soulignent les réponses qui peuvent être apportées à ces problèmes, en particulier en revoyant les techniques de recensement appliquées jusqu’à présent pour évaluer les populations de chiens errants, ce qui permettra ensuite de faciliter la planification et la conception des programmes de vaccination. En outre, ils examinent les différentes stratégies de vaccination mises en oeuvre en la matière et font le point sur les moyens d’évaluer la couverture vaccinale de ces populations. Enfin, ils analysent les avantages apportés par les programmes de stérilisation pour maîtriser les populations de chiens errants. En résumé, cet article met l’accent sur l’importance d’inclure les chiens errants au même titre que les chiens ayant un maître dans les programmes de vaccination et souligne que la plupart des difficultés liées à la protection des chiens errants contre la rage peuvent être résolues.Uno de los aspectos capitales de todo programa de vacunación antirrábica es la gestión eficaz de las poblaciones de perros vagabundos. En muchos países con rabia endémica, estos perros representan un porcentaje sustancial tanto de la población canina total como de los perros en libertad. El éxito de todo programa de eliminación de la rabia pasa necesariamente por lograr una elevada cobertura de vacunación en este segmento de la población canina (los perros vagabundos). Sin embargo, la gestión y la vacunación antirrábica de los perros vagabundos presentan una serie de dificultades. Los autores pasan revista a los aspectos que más inciden en el problema de los perros vagabundos y la eliminación de la rabia, prestando especial atención a las dificultades que presentan la estimación del tamaño de la población, la administración de vacunas y el cálculo de la cobertura de vacunación de las poblaciones de perros vagabundos. Después explican la manera de superar estas dificultades, en particular describiendo las técnicas de censo empleadas para calcular el tamaño de las poblaciones, lo que a su vez facilita la concepción y planificación de programas de vacunación. Además, describen el arsenal existente de procedimientos de vacunación que se han utilizado para vacunar a los perros vagabundos y explican cómo estimar en estas poblaciones la cobertura de vacunación. Por último exponen los beneficios de gestionar las poblaciones de perros vagabundos con programas de esterilización. En resumidas cuentas, los autores recalcan aquí la importancia de que los programas de vacunación canina lleguen a los perros vagabundos, y no solo a los animales con dueño, e insisten en que muchos de los problemas que se plantean para proteger de la rabia a los perros vagabundos tienen solución.

This focus article has been prepared by

The combination of soft nanoscale organic components with inorganic nanograins hierarchically designed by natural organisms results in highly ductile structural materials that can withstand mechanical impact and exhibit high resilience on the macro- and nano-scale. Our investigation of nacre deformation reveals the underlying nanomechanics that govern the structural resilience and absorption of mechanical energy. Using high-resolution scanning/transmission electron microscopy (S/TEM) combined with in situ indentation, we observe nanoscale recovery of heavily deformed nacre that restores its mechanical strength on external stimuli up to 80% of its yield strength. Under compression, nacre undergoes deformation of nanograins and non-destructive locking across organic interfaces such that adjacent inorganic tablets structurally join. The locked tablets respond to strain as a continuous material, yet the organic boundaries between them still restrict crack propagation. Remarkably, the completely locked interface recovers its original morphology without any noticeable deformation after compressive contact stresses as large as 1.2 GPa., 4 figures

Complementation-dependent fluorescence is a powerful way to study co-localization or interactions between biomolecules. A split-GFP variant, involving the self-associating GFP 1–10 and GFP 11, has previously provided a convenient approach to measure recombinant protein titers in cell supernatants. A limitation of this approach is the slow chromophore formation after complementation. Here, we alleviate this lag in signal generation by allowing the GFP 1–10 chromophore to mature on a solid support containing GFP 11 before applying GFP 1–10 in analyses. The pre-maturated GFP 1–10 provided up to 150-fold faster signal generation compared to the non-maturated version. Moreover, pre-maturated GFP 1–10 significantly improved the ability of discriminating between Chinese hamster ovary (CHO) cell lines secreting GFP 11-tagged erythropoietin protein at varying rates. Its improved kinetics make the pre-maturated GFP 1–10 a suitable reporter molecule for cell biology research in general, especially for ranking individual cell lines based on secretion rates of recombinant proteins.

We recently investigated an outbreak of botulism in a duck ( Anas platyrhynchos ) and pheasant ( Phasianus colchicus ) farm where 30,000 mallard ducks and 35,000 pheasants are reared from one day old for shooting. The farm set up is that ducks are kept on several ponds with the water refreshed using pumps. The pheasants are initially reared in small pens and later in large runs with adjacent cover provided by maize. The birds each receive commercial pelleted feed. From the middle of August increased losses were identified in the ducks on one of the ponds and in the pheasants in one of the runs, which was nearest to the affected pond. Ducks also wandered into this run. The ducks and pheasants were reported to show similar clinical signs: weakness, inability to stand, becoming recumbent on their keels, with wings extended and resting their beaks on the ground. Six dead mallard ducks and two live pheasants were submitted for examination to the APHA Shrewsbury Veterinary Investigation Centre. Five of the ducks had been …

Wounds in working donkeys are a common and preventable welfare problem in many countries. Mutilations, iatrogenic injuries carried out by owners, are a particularly distressing welfare issue. However, little is known about their nature and prevalence. The project aim was to assess the general health and establish the prevalence, nature and severity of mutilations and other skin wounds in donkeys from several communities in Tamil Nadu, India, that were part of a donkey welfare initiative run by the Worldwide Veterinary Service charity. Five hundred and eighty-two donkeys were examined from five locations and data collected using a predesigned, smartphone app between October 2016 and July 2017. The study revealed that 298 wounds were present in a total of 227 donkeys (39.0 per cent of the population). Mutilations, comprising nose-splitting, ear-splitting and branding, were the most common type of skin wound, comprising 62.8 per cent of all wounds. Poorly fitting harnesses and hobbles were also a common cause of injury. These data can be used to design future-targeted and focused, educational interventions to reduce mutilation practices which are a common and important welfare issue. Further outreach initiatives are urgently needed to improve welfare of donkeys in these regions.

Biomagnification of methylmercury in aquatic systems can cause elevated tissue mercury (Hg) and physiological stress in top predators. Mercury is known to affect stress hormone levels in mammals, birds and fish. In this study, the effects of maternally-transferred methylmercury on the stress physiology of Northern Water Snake (Nerodia sipedon) neonates were tested. Gravid females were dosed via force-fed capsules during late gestation with 0, 0.01, or 10 µg methylmercury per gram of body mass. Plasma corticosterone levels and leukocyte differentials were analyzed in baseline and confinement-stressed neonates from all dose levels. Neither Hg nor confinement stress had a significant effect on leukocyte differentials nor was Hg related to corticosterone levels. However, stress group neonates showed lower heterophil/lymphocyte ratios and this study was the first to show that neonate N. sipedon can upregulate CORT in response to stress. These results indicate that N. sipedon may be somewhat tolerant to Hg contamination.