One Health: Animal Models of Heritable Human Bleeding Diseases.

Animal models of human and animal diseases have long been used as the lynchpin of experimental and clinical research. With the discovery and implementation of novel molecular and nano-technologies, cellular research now has advanced to assessing signal transduction pathways, gene editing, and gene therapies. The contribution of heritable animal models to human and animal health as related to hemostasis is reviewed and updated with the advent of gene editing, recombinant and gene therapies.

Early Life Vaccination of Companion Animal Pets.

Development of the immune system of mammalian animal species parallels that of humans and involves the innate and adaptive (acquired) immune responses acting together with the thymus gland. Consequently, issues surrounding the adequacy and safety of vaccinations to protect pet animals from their relevant infectious diseases need to be addressed just as they are for humans. Pet animals, especially canines, also have unique needs because of the wide diversity of purebred and mixed breeds that vary greatly in size, type, temperament, and even maturation rates. Furthermore, pets in early life encounter a series of changes that can affect their development and induce stressors including parasite control, new homes and environment, novel foods, and the socialization that is essential at a time when vaccinations need to be given. While recognizing that this overall need is becoming more understood, current vaccination policy guidelines for companion animals are still only adhered to by about 40% of veterinarians worldwide. Clearly, vaccination of pets should no longer be considered as "one size fits all".

Survey of canine use and safety of isoxazoline parasiticides.

A veterinarian and pet owner survey (Project Jake) examined the use and safety of isoxazoline parasiticides given to dogs. Data were received during August 1-31, 2018 from a total of 2,751 survey responses. Forty-two percent (1,157) reported no flea treatment or adverse events (AE), while 58% (1594) had been treated with some parasiticide for flea control, and of those that received a parasiticide, the majority, or 83% (1,325), received an isooxazoline. When any flea treatment was given, AE were reported for 66.6% of respondents, with no apparent AE noted for 36.1%. Project Jake findings were compared to a retrospective analysis of publicly available Food and Drug Administration (FDA) and European Medicines Agency (EMA) reported AE. The number of total AE reported to FDA and EMA were comparable, although a 7 to 10 times higher occurrence of death and seizures was reported from the EMA or from outside the United States (US). Serious AE responses for death, seizures and neurological effects reported in our survey were higher than the FDA but moderately lower than the EMA reports. These sizable global data sets combined with this pre- and post-parasiticide administration survey indicated that isoxazoline neurotoxicity was not flea- and tick-specific. Post-marketing serious AE were much higher than in Investigational New Drug (IND) submissions. Although the labels have recently been updated, dogs, cats and their caregivers remain impacted by their use. These aggregate data reports support the need for continued cross-species studies and critical review of product labelling by regulatory agencies and manufacturers.

Duration of immunity after rabies vaccination in dogs: The Rabies Challenge Fund research study.

A prospective study of 65 research beagles kept in a rabies-free environment was undertaken to determine the duration of immunity after they received licensed rabies vaccines. The eventual goal was to extend mandated rabies booster intervals to 5 or 7 years and help reduce the risk of vaccine-associated adverse events. Three groups of dogs were vaccinated with 1 of 2 commercial rabies vaccines or saline at 12 and 15 weeks of age. Beginning 5 years 5 months later, vaccinated and unvaccinated dogs were challenged with virulent rabies virus and observed for 90 days over a series of 3 trials. Humoral and cellular immune responses were examined by serology and flow cytometry. Brain tissue from all challenged dogs was tested for rabies virus. Challenge trial 1 was confounded due to insufficiently virulent virus. In trials 2 and 3 virulent challenge provided 100% mortality in controls. Vaccinate survival was 80% (4/5) after 6 years 7 months, 50% (6/12) after 7 years 1 month, and 20% (1/5) after 8years 0 months. Antibody responses 12 days post-challenge correlated strongly with survival. In a separate non-challenge trial, administration of either a recombinant or a killed rabies vaccine demonstrated memory antibody responses 6 years 1 month after initial vaccination compared with unvaccinated controls. Our data demonstrated that i) duration of immunity to rabies in vaccinated dogs extends beyond 3 years; ii) immunologic memory exists even in vaccinated dogs with serum antibody titer < 0.1 IU/mL; and iii) non-adjuvanted recombinant rabies vaccine induces excellent antibody responses in previously vaccinated dogs 14 days after administration.

Une étude prospective sur 65 chiens beagle de recherche gardés dans un environnement exempt de rage fut entreprise afin de déterminer la durée de l'immunité après qu'ils reçurent un vaccin homologué contre la rage. Le but éventuel était d'allonger l'intervalle requis du rappel du vaccin contre la rage à 5 ou 7 ans et aider à réduire le risque associé aux réactions adverses au vaccin. Trois groupes de chiens furent vaccinés avec un des deux vaccins commerciaux contre la rage ou de la saline à 12 et 15 semaines d'âge. Débutant 5 ans et 5 mois plus tard, les chiens vaccinés et non-vaccinés furent soumis à une infection défi avec un virus de la rage virulent et observés pendant 90 jours lors d'une série de trois essais. Les réponses immunitaires humorale et cellulaire furent examinées par sérologie et cytométrie de flux. Du tissu cérébral de tous les chiens infectés fut testé pour la présence du virus rabique. L'essai 1 était décevant étant donné la quantité insuffisante de virus virulent. Lors des essais 2 et 3, l'infection défi a entrainé 100 % de mortalité chez les témoins. Le taux de survie des animaux vaccinés était de 80 % (4/5) après 6 ans et 7 mois, 50 % (6/12) après 7 ans et 1 mois et 20 % (1/5) après 8 ans 0 mois. La réponse en anticorps 12 jours post-infection corrélait fortement avec la survie. Dans un essai séparé sans infection défi, l'administration de soit un vaccin recombinant ou un vaccin tué a mis en évidence une réponse anamnestique en anticorps 6 ans et 1 mois après la vaccination initiale comparativement aux témoins non-vaccinés. Nos résultats démontrent que (1) la durée de l'immunité contre la rage chez les chiens vaccinés va au-delà de 3 ans, (2) une mémoire immunologique existe même chez les chiens vaccinés avec des titres d'anticorps sériques < 0,1 IU/mL, et (3) un vaccin antirabique recombinant sans adjuvant induit d'excellentes réponses en anticorps chez des chiens préalablement vaccinés 14 jours après son administration.(Traduit par Docteur Serge Messier).

Diagnosis of Feline Food Sensitivity and Intolerance Using Saliva: 1000 Cases.

This prospective study assessed the efficacy of a novel saliva-based immunoassay of IgA- and IgM-antibodies in predicting feline food sensitivities and intolerances. Clinical samples were obtained from 1000 cats proven or suspected to have food intolerances. Most were of domestic shorthair breed type, over 10 years of age, and weighed around 5 kg; they were equally distributed between spayed females and neutered males. Saliva was collected after at least an 8-h fast with a dental cotton rope, placed in a double-sleeved saliva collection tube, and sent to the laboratory. Salivary antibodies elicited by 24 common foods were measured with goat anti-canine IgA and IgM. Low reacting foods were lamb, cow milk, pork, turkey, wheat (lowest) and white-colored fish, whereas high reacting foods were millet, white potato, rice (highest) and salmon. Thus, the novel salivary-based food sensitivity and intolerance test, described previously for canines, also provided a reliable and clinically predictive alternative to food elimination trials, serum-based food allergy testing, and skin patch testing in cats. Manufacturers of commercial cat foods and treats, as well as those making homemade diets and treats for cats, should consider avoiding the more highly reactive foods as determined by the present study.

Testing for food-specific antibodies in saliva and blood of food allergic and healthy dogs.

An elimination diet (ED) followed by re-challenge has been the reference standard to diagnose adverse food reactions (AFR) in dogs, but can be challenging to conduct. This study investigated the accuracy of a saliva-based test for food-specific IgA and IgM and an ELISA serum test for food-specific IgE. Three groups of dogs were tested. Group 1 (n=11) included dogs with previously diagnosed and controlled AFR; group 2 (n=15) comprised dogs with allergic dermatitis at the beginning of their ED; and group 3 (n=16) was composed of clinically healthy research dogs. Saliva samples were collected from all groups and blood samples from group 1 and group 3. The results of clinical re-challenges with individual food components were compared with the test results. Specificity, sensitivity, positive and negative predictive values and likelihood ratios were determined. Forty-one dogs completed the study; one dog was lost to follow up. There was a total of 163 re-challenges. Sensitivity, positive predictive value and likelihood ratio, specificity, negative predictive value and likelihood ratios were unsatisfactory for both tests in most instances, except for IgM testing in group 2, which had moderate specificity. There was no clear difference in the number of positive reactions between the allergic dogs and healthy dogs from a research population. Based on these results, the saliva test for food specific IgA and IgM and the ELISA serum test for food specific IgE were not reliable to diagnose adverse food reactions in dogs. Until more data are available, elimination diets remain the reference standard in the diagnosis of this disease.

Relationships Between Land Use and Stream Nutrient Concentrations in a Highly Urbanized Tropical Region of Brazil: Thresholds and Riparian Zones.

Nutrient enrichment in streams due to land use is increasing globally, reducing water quality and causing eutrophication of downstream fresh and coastal waters. In temperate developed countries, the intensive use of fertilizers in agriculture is a main driver of increasing nutrient concentrations, but high levels and fast rates of urbanization can be a predominant issue in some areas of the developing world. We investigated land use in the highly urbanized tropical State of Rio de Janeiro, Brazil. We collected total nitrogen, total phosphorus, and inorganic nutrient data from 35 independent watersheds distributed across the State and characterized land use at a riparian and entire watershed scales upstream from each sample station, using ArcGIS. We used regression models to explain land use influences on nutrient concentrations and to assess riparian protection relationships to water quality. We found that urban land use was the primary driver of nutrient concentration increases, independent of the scale of analyses and that urban land use was more concentrated in the riparian buffer of streams than in the entire watersheds. We also found significant thresholds that indicated strong increases in nutrient concentrations with modest increases in urbanization reaching maximum nutrient concentrations between 10 and 46% urban cover. These thresholds influenced calculation of reference nutrient concentrations, and ignoring them led to higher estimates of these concentrations. Lack of sewage treatment in concert with urban development in riparian zones apparently leads to the observation that modest increases in urban land use can cause large increases in nutrient concentrations.

Remineralization of early caries by chewing sugar-free gum: a clinical study using quantitative light-induced fluorescence.

PURPOSE: To determine whether sugar-free gum can provide remineralization and caries control of active enamel caries lesions compared to baseline (before gum chewing) and to a no-gum group, following daily chewing for 12 weeks by school children; to determine whether chewing frequency can affect the extent of remineralization. METHOD: A pragmatic cluster-randomized controlled clinical trial with schools as the unit of randomization was employed. Three schools in Chengdu, PR China comprised the clusters. The study was approved by the Internal Review Board of Sichuan University. 177 school children, 8-13 years old, with at least one visible white-spot lesion were enrolled in the study. Each of the three clusters was randomly assigned to one of three groups: (1) no gum; (2) chew 2 pieces of sugar-free gum for 20 minutes, 3x per day; (3) chew 2 pieces of sugar-free gum for 12 minutes, 5x per day. White-spot lesions were examined by quantitative light-induced fluorescence (QLF) at baseline and after 4, 8, and 12 weeks of treatment. RESULTS: 155 subjects completed the study. Of them, the mean values of fluorescence loss at baseline were 9.52, 9.83 and 9.17 for no-gum group, 3x per day group and 5x per day group, respectively. For the area, the mean values at baseline were 2.52, 2.61 and 2.57 mm2 for no-gum group, 3x per day group and 5x per day group, respectively. For AQ, the mean values at baseline were -27.91, -28.29 and -29.67 for no-gum group, 3x per day group and 5x per day group, respectively. To adjust for differences in groups at baseline, ANCOVA was used. After 12-weeks, for all QLF metrics, the absolute values of 5x per day group were the lowest and the no gum group was the highest; the differences among three groups were statistically significant (P < 0.05). For AQ, which was accepted as the most useful metrics of QLF system, the adjusted mean values at 12 weeks were -26.35, -19.81 and -17.58 for no-gum group, 3x per day group and 5x per day group, respectively. There were significant differences between groups.

Serological evidence of H3N8 canine influenza-like virus circulation in USA dogs prior to 2004.

H3N8 canine influenza virus (H3N8 CIV) was first reported as a novel canine respiratory pathogen in racing greyhounds and shelter dogs in the U.S.A. in 2004. Phylogenetic analyses determined that this host-adapted pathogen originated from interspecies transmission of an equine influenza virus (EIV), but it is unknown when the transmission occurred prior to discovery in 2004. The objective of this study was to determine if racing greyhound and shelter dog sera collected from 1984 to 2004 had serological evidence of exposure to H3N8 CIV or EIV. Archived sera from 702 racing greyhounds and 1568 shelter dogs were tested for H3 antibodies to the original 2004 CIV isolate, as well as EIV isolates from 1991 to 1999. None of the racing greyhounds from 1984 and 1985 had detectable H3 antibodies. One of the shelter dogs, which entered a north Florida shelter in 2004, was seropositive. For racing greyhounds sampled from 1999 to 2004, 133/520 (26%) dogs had antibodies to both CIV and EIV H3 proteins. The annual seroprevalence was 27% in 1999, 28% in 2000, 10% in 2001, 1% in 2002, 41% in 2003, and 28% in 2004. The odds of H3 seropositivity were greater among dogs that raced > or =6 months, raced on > or =2 tracks, and raced in 1998, 2002, and 2003. Many of the seropositive dogs raced at tracks that were involved in 'kennel cough' epidemics in 1998-1999 and 2002-2003. Based on serological evidence, a H3N8 canine influenza-like virus was circulating in racing greyhounds in the U.S.A. as early as 1999.

Association of canine hypothyroidism with a common major histocompatibility complex DLA class II allele.

Dogs exhibit a range of immune-mediated conditions including a lymphocytic thyroiditis which has many similarities to Hashimoto's thyroiditis in man. We have recently reported an association in Doberman Pinschers between canine hypothyroidism and a rare DLA class II haplotype that contains the DLA-DQA1*00101 allele. We now report a further series of 173 hypothyroid dogs in a range of breeds where a significant association with DLA-DQA1*00101 is shown.

Implementing the 2-week wait rule for cancer referral in the UK: general practitioners' views and practices.

The 2-week wait rule for cancer referrals became effective in December 2000 for all cancers treated by the National Health Service in the UK. Attainment of this target depends initially on appropriate and timely referral by general practitioners (GPs). General practitioners' views and referral practices under the 2-week wait rule were examined based on a postal survey of 508 GPs in an inner London area (65% response). Data on mode of referral indicated that 90% of GPs used the urgent suspected cancer form, although 38% also sent a letter with further information. General practitioners generally regarded the 2-week wait rule as working well in terms of improving patients' initial access, and 50% thought communication with the hospital had improved. However, 46% expressed some concerns, including problems arising from the different sets of forms required by local cancer networks, the lack of a dedicated referral form for breast cancer, and feelings of a loss of autonomy. General practitioners also acknowledged an element of over-referral under this rule due to the effects of clinical uncertainty and patient pressure and their concerns about increased waiting times for non-target referrals. The survey therefore indicates that GPs are generally positive about the 2-week wait rule but identified some problems of implementation including a need for standardized national cancer referral forms.