VetBiobank, Vetmeduni Vienna: A bioresource for clinical animal biospecimens

EXECUTIVE SUMMARYIn 2012 and 2013, the World Small Animal Veterinary Association (WSAVA) Vaccination Guidelines Group (VGG) undertook fact‐finding visits to several Asian countries, with a view to developing advice for small companion animal practitioners in Asia related to the administration of vaccines to dogs and cats. The VGG met with numerous first opinion practitioners, small animal association leaders, academic veterinarians, government regulators and industry representatives and gathered further information from a survey of almost 700 veterinarians in India, China, Japan and Thailand. Although there were substantial differences in the nature and magnitude of the challenges faced by veterinarians in each country, and also differences in the resources available to meet those challenges, overall, the VGG identified insufficient undergraduate and postgraduate training in small companion animal microbiology, immunology and vaccinology. In most of the countries, there has been little academic research into small animal infectious diseases. This, coupled with insufficient laboratory diagnostic support, has limited the growth of knowledge concerning the prevalence and circulating strains of key infectious agents in most of the countries visited.Asian practitioners continue to recognise clinical infections that are now considered uncommon or rare in western countries. In particular, canine rabies virus infection poses a continuing threat to animal and human health in this region. Both nationally manufactured and international dog and cat vaccines are variably available in the Asian countries, but the product ranges are small and dominated by multi‐component vaccines with a licensed duration of immunity (DOI) of only 1 year, or no description of DOI. Asian practitioners are largely unaware of current global trends in small animal vaccinology or of the WSAVA vaccination guidelines. Consequently, most practitioners continue to deliver annual revaccination with both core and non‐core vaccines to adult animals, with little understanding that “herd immunity” is more important than frequent revaccination of individual animals within the population.In this paper, the VGG presents the findings of this project and makes key recommendations for the Asian countries. The VGG recommends that (1) Asian veterinary schools review and increase as needed the amount of instruction in small animal vaccinology within their undergraduate curriculum and increase the availability of pertinent postgraduate education for practitioners; (2) national small animal veterinary associations, industry veterinarians and academic experts work together to improve the scientific evidence base concerning small animal infectious diseases and vaccination in their countries; (3) national small animal veterinary associations take leadership in providing advice to practitioners based on improved local knowledge and global vaccination guidelines; (4) licensing authorities use this enhanced evidence base to inform and support the registration of improved vaccine product ranges for use in their countries, ideally with DOI for core vaccines similar or equal to those of equivalent products available in western countries (i.e. 3 or 4 years).The VGG also endorses the efforts made by Asian governments, non‐governmental organisations and veterinary practitioners in working towards the goal of global elimination of canine rabies virus infection. In this paper, the VGG offers both a current pragmatic and future aspirational approach to small animal vaccination in Asia. As part of this project, the VGG delivered continuing education to over 800 Asian practitioners at seven events in four countries. Accompanying this document is a list of 80 frequently asked questions (with answers) that arose during these discussions. The VGG believes that this information will be of particular value to Asian veterinarians as they move towards implementing global trends in small companion animal vaccinology.

Live-attenuated bacterial veterinary vaccines can constitute an infection risk for individuals with any defect in their phagocytic function, including chronic granulomatous disease, leukocyte adhesion deficiency, myeloperoxidase deficiency, as well as Chediak-Higashi syndrome, from accidental acquisition of licenced attenuated live bacterial vaccine, at vaccination or from their vaccinated pet. Ownership of small companion animals, including cats and dogs, is popular within the cystic fibrosis (CF) community. These animals require vaccines as part of their routine care, which may involve live viral and bacterial vaccines, with potential for infection in the CF owner. This report examines the scope of current canine and feline vaccines, with particular emphasis on veterinary vaccination strategies against the Gram-negative pathogen, Bordetella bronchiseptica and describes new vaccine innovations offering protection to both pet and CF owner. The Gram-negative bacterium, Bordetella bronchoseptica, may cause respiratory disease in small companion animals, as well as in certain human vulnerable groups, including those with CF. Live licenced veterinary bacterial vaccines for Bordetella bronchiseptica (Kennel Cough) are available for cats and dogs, which are an infection concern for humans with CF who may come into contact with vaccinated animals. Live licenced veterinary bacterial vaccines for Bordetella bronchiseptica (Kennel Cough) are available for intranasal administration to cats and dogs. These vaccines require a withdrawal period of vaccinated animal from vulnerable owner, ranging from 35days - 11weeks. Recently, a new dead IM vaccine is now available not requiring exclusion of the vaccinated pet from CF owner. CF pharmacists, hospital pharmacists and community pharmacists are important custodians of vaccine-related advice to people with CF, who are frequently consulted for such advice. Pharmacists should be aware of the recent innovations in veterinary medicines, so that they can give appropriate advice to people with CF when asked. Immunocompromised patients, that is those with CF or those with any defect in their phagocytic function (chronic granulomatous disease, leukocyte adhesion deficiency, myeloperoxidase deficiency, Chediak-Higashi syndrome) should avoid exposure to live veterinary bacterial vaccines and seek animal vaccination utilising non-live vaccines. Most importantly, this manuscript highlights the development of a new veterinary vaccine for dogs, which we want to make the CF healthcare community aware of, which is an acellular dead vaccine, so that those patients with dogs needing annual vaccination can select this vaccine pathway, thereby minimising risk of infection from the vaccine strains and avoiding the social exclusion between CF patient and their pet. CF patients should understand the potential infection implications of live-attenuated viral and bacterial strains as vaccines, whether these are small companion animals, exotic animals or large farm animals. Patients should make their veterinarian aware of their CF status, so that a safe and efficacious vaccine strategy is used, both mitigating the potential infection risks from live vaccine components with the CF patient, but simultaneously offering maximum immunological protection to the animal.

ABSTRACTRecent years have seen significant developments in the ability to continuously propagate organoids derived from intestinal crypts. These advancements have been applied to mouse and human samples providing models for gastrointestinal tissue development and disease. We adapt these methods for the propagation of intestinal organoids (enteroids) from various large farm and small companion (LF/SC) animals, including cat, dog, cow, horse, pig, sheep and chicken. We show that LF/SC enteroids propagate and expand in L-WRN conditioned media containing signaling factors Wnt3a, R-spondin-3, and Noggin (WRN). Multiple successful isolations were achieved for each species, and the growth of LF/SC enteroids was maintained to high passage number. LF/SC enteroids expressed crypt stem cell marker LGR5 and low levels of mesenchymal marker VIM. Labeling with EdU also showed distinct regions of cell proliferation within the enteroids marking crypt-like regions. The ability to grow and maintain LF/SC enteroid cell lines provides additional models for the study of gastrointestinal developmental biology as well as platforms for the study of host-pathogen interactions between intestinal cells and zoonotic enteric pathogens of medical importance.

The International Society for Biological and Environmental Repositories Presents Abstracts from Its Annual Meeting Due North: Aligning Biobanking Practice with Evolving Evidence and Innovation May 9–12, 2017 Toronto, Canada

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This paper examines the turn toward the small companion animal that occurred in British veterinary medicine in the twentieth century. The change in species emphasis is usually attributed to post-war socioeconomic factors, however this explanation ignores the extensive small animal treatment that was occurring outwith the veterinary profession in the interwar period. The success of this unqualified practice caused the veterinary profession to rethink attitudes to small animals (dogs initially, later cats) upon the decline of horse practice. This paper argues that a shift toward seeing the small animal as a legitimate veterinary patient was necessary before the specialty could become mainstream in the post-war years, and that this occurred between the wars as a result of the activities of British animal welfare charities, especially the People's Dispensary for Sick Animals of the Poor.

Antimicrobial Resistance

Spontaneous mammary tumors are very common in bitches. The involvement of oxidative stress and the function of antioxidant enzymes in cancerogenesis have been studied in depth in human medicine, while data in veterinary medicine are still fragmentary. The main aim of this study was to evaluate the activity and the expression of superoxide dismutases (Cu-ZnSOD and MnSOD) and the activity of catalase (CAT) in canine mammary tumors in comparison with the adjacent healthy tissue. Six female dogs (mean age 10.4 years) were included in this study. After surgery, fresh tumor and healthy tissue samples were immediately frozen in dry ice and stored at −80°C for biochemical analyses, while the remaining parts were used for histopathological analysis. Enzyme activity was measured by spectrophotometric assays and protein expression by western blotting. In canine mammary tumors, Cu-ZnSOD activity and expression increased significantly compared with healthy control tissues (p=0.03). MnSOD showed a significantly lower activity in tumoral tissues at stage 2 (p<0.05), while a significant increase of expression was measured in tumors. CAT activity was significantly higher in healthy tissues respect to tumors (p=0.015). These variations of antioxidant enzymes activities and expression could be related to an increase of oxidative stress in breast cancer tissues and could be considered as biomarker candidates for neoplastic transformation.

Vaccination against infectious diseases is a cornerstone of veterinary medicine in the prevention of disease transmission, illness severity, and often death in animals. In North American equine medicine, equine vaccines protecting against tetanus, rabies, Eastern and Western equine encephalomyelitis, and West Nile are core vaccines as these have been classified as having a heightened risk of mortality, infectiousness, and endemic status. Some guidelines differ from the label of vaccines, to improve the protection of patients or to decrease the unnecessary administration to reduce potential side effects. In North America, resources for the equine practitioners are available on the American Association of Equine Practitioners (AAEP) website. Conversely, in small companion animals, peer review materials are regularly published in open access journals to guide the vaccination of dogs and cats. The aims of this review are to present how the vaccine guidelines have been established for small companion animals and horses in North America, to review the equine literature to solidify or contrast the current AAEP guidelines of core vaccines, and to suggest future research directions in the equine vaccine field considering small companion animal strategies and the current available resources in equine literature.

Research-based biobanks will create new synergies between industry and public research structures, strengthening the competitiveness of our country for health industries. In addition to the ultimate objective of prevention and treatment of complex diseases, the short-term benefit will come from the development of new and more powerful diagnostic agents. In fact, molecular diagnostics, a new discipline based on “-omics” technologies, a powerful tool to understand diseases and assist individuals at risk, is one of the fastest growing segments in the healthcare industry. In order to collect the socio-economic benefits summarised above, many European countries, have underway, or are planning, large, well organised biobanks. However, the existing samples collection, resources, technologies and expertise have been developed under different ethical and legal landscapes across Europe. The existing collections suffer from fragmentation, variable access rules and the lack of commonly applied standards. This lack of standardization prevents the effective use of biological samples and data from different biobanks, which is a prerequisite to achieve sufficient statistical power for genomics research in humans. By initiating the Biobanking and Biomolecular Research Infrastructure (BBMRI) [All Member States and the Commission agreed on a shared vision of how the European Research Area (ERA) should develop by 2020 (http://ec.europa.eu/research/era/index_en.htm). This vision was adopted by the Council of Ministers in December 2008, and implemented for the needed infrastructures by ESFRI, the European Strategy Forum on Research Infrastructures, a already existing strategic instrument with the task of developing the scientific integration of Europe and to strengthen its international outreach. Since it was formed in 2002, at the behest of the European Council, ESFRI has witnessed significant advances towards unity and international impact in the field of research infrastructures. The publication of the first Roadmap for pan-European research infrastructures in 2006, and its update in 2008 (http://ec.europa.eu/research/infrastructures/index_en.cfm?pg=esfri) was a key contributing factor, and several projects are now in the realization phase like BBMRI (http://www.bbmri.eu/). BBMRI is a large scale project building a research infrastructure structured as a federated network of biological resources centres including all types of biobanks: disease oriented, population-based biobanks, biomolecular-resources; it includes all fields of medical research: rare diseases, cancer, complex diseases, etc.] on February 1st, 2008, Europe addressed the need to: harmonise standards for sample collection, storage and analysis; harmonise data collection and database infrastructure; provide ethical and legal guidance; develop a sustainable funding model for biobanks. This article describes some of the aspects of this European infrastructure.

Breast cancer is one of the most common cancers known among women. This study aimed to investigate the level of vitamin D receptor gene expression in two tumoral and healthy breast tissues in breast cancer patients and its association with prognostic factors. This descriptive cross-sectional study was conducted in 2022 on 50 patients with high suspicion of breast cancer who were candidates for mastectomy and lumpectomy in a learning hospital. From the patients, two tissue samples were prepared, and there was a total of 100 samples. The samples were subjected to H/E staining and evaluated by a pathologist. The presence or absence of malignancy in each sample was confirmed by two pathologists, and HER2/ER/PR indices were determined. Descriptive and analytical statistical methods and SPSS version 22 software were used. The average age of the patients was 51.60 ± 11.22 years old, and the average tumor size was 3.17 ± 1.28. Most tumors were grade 2 (48%). The expression of HER2, ER, and PR was positive in 24, 64, and 54%, respectively. The largest number of cases were in stage 2A. The expression level of vitamin D receptor (VDR) gene in healthy tissue (2.08 ± 1.01) was higher than tumoral tissue (0.25 ± 1.38) (P = 0.001). In tumoral and healthy tissue, VDR expression was not significant according to tumor grade, HER2, ER, PR, LVI, LN, disease stage, age, and tumor size. The expression level of VDR in healthy tissue was significantly higher than tumoral tissue. However, there was no significant relationship between VDR and tumor grade, HER2, ER, PR, LVI, LN, disease stage, age, and tumor size.

California wildfires are expected and they are followed by landslides and floods. A changing culture in traditional responders has co-evolved with a culture of pet owner disaster preparedness in California. This is demonstrated by the documentation of the in California wildfires from 2003–2009. The 2003 wildfires in San Diego County involved a massive relocation of people, small companion animals and over 700 horses. It was a milestone event in allowing co-located human and animal shelters. The intent was for animals to be maintained in adjacent shelters, but the result was humans taking up residence with their animal companions. Pet owners sought to keep their family, including their pets, together. It appeared that pets were providing comfort to their owners. In the same region during 2007, pet owners mobilized rapidly. They were clearly more prepared than they were in 2003. During 2008, a record 1400 fires were burning in California on July 1st. The Santa Barbara Gap Fire mobilized an expert Santa Barbara Equine evacuation team. The human Red Cross evacuation center allowed small animal cohabitation. At the same time, the Butte Lightning Complex fires (37 fires) involved a unique cohort of canine evacuees for an extended duration and requiring unprecedented veterinary volunteers. In 2009, the Santa Barbara Jesusita fire threatened an urban area and evacuation of 35,000 people. It included a vulnerable human population with health disabilities that required ambulance evacuation assistance. Small companion animals were allowed to evacuate in the front cab of the ambulance. Ambulance drivers remarked that they dreaded forcing patients to leave behind their pets and it was a relief to bring the pets along. In summary, the response to repeated California wildfires from 2003 to 2009 has demonstrated an evolving culture of animal disaster preparedness for both traditional responders and companion animal owners.

BackgroundClostridium difficile infections (CDI) in humans range from asymptomatic carriage to life-threatening intestinal disease. Findings on C. difficile in various animal species and an overlap in ribotypes (RTs) suggest potential zoonotic transmission. However, the impact of animals for human CDI remains unclear.MethodsIn a large-scale survey we collected 1,447 fecal samples to determine the occurrence of C. difficile in small companion animals (dogs and cats) and their owners and to assess potential epidemiological links within the community. The Germany-wide survey was conducted from July 2012-August 2013. PCR ribotyping, Multilocus VNTR Analysis (MLVA) and PCR detection of toxin genes were used to characterize isolated C. difficile strains. A database was defined and logistic regression used to identify putative factors associated with fecal shedding of C. difficile.ResultsIn total, 1,418 samples met the inclusion criteria. The isolation rates for small companion animals and their owners within the community were similarly low with 3.0% (25/840) and 2.9% (17/578), respectively. PCR ribotyping revealed eight and twelve different RTs in animals and humans, respectively, whereas three RTs were isolated in both, humans and animals. RT 014/0, a well-known human hospital-associated lineage, was predominantly detected in animal samples. Moreover, the potentially highly pathogenic RTs 027 and 078 were isolated from dogs. Even though, C. difficile did not occur simultaneously in animals and humans sharing the same household. The results of the epidemiological analysis of factors associated with fecal shedding of C. difficile support the hypothesis of a zoonotic potential.ConclusionsMolecular characterization and epidemiological analysis revealed that the zoonotic risk for C. difficile associated with dogs and cats within the community is low but cannot be excluded.

Purpose: The purpose of this pilot study was to assess the differences in breast tissue microbiota composition by breast side (left versus right) within an individual woman and compare the microbiota of healthy and breast tumor tissue between women. We further aimed to determine whether certain bacterial taxa may be associated with breast tumors. Methods: Bilateral healthy breast tissue samples (n=36) were collected from ten women who received routine mammoplasty procedures at the University of Florida Department of Surgery. Archived breast tumor samples (n=10) were obtained from an established biorepository. Bacterial DNA was extracted from tissues, amplified via polymerase chain reaction (PCR), and sequenced on the Illumina MiSeq platform. Microbiota data were analyzed using QIIME and RStudio. Results: The most abundant phyla in both tumor and healthy tissues were Bacteroidetes, Firmicutes, Proteobacteria, and Actinobacteria. A total of 38 operational taxonomic units (OTUs) were found to be significantly different in terms of differential abundance between tumor and healthy tissues (absolute effect size range: 0.761-3.98). The OTUs with the largest absolute effect size associated with higher relative abundance in breast tumors were of Flavobacterium species (R2=3.98), Acinetobacter species (R2=3.64), [Mogibacteriaceae] family (R2=3.34), and Clostridiales order (R2=3.21). Alpha diversity (Shannon Diversity Index) was similar in healthy and tumor tissue (4.98 vs. 4.84; p=0.350). Based on unweighted UniFrac measures, breast tumor samples clustered distinctly from healthy samples (R2=0.10; p=0.001). Microbiota composition in healthy samples clustered within women (R2=0.20; p=0.012) and by breast side (left or right) within a woman (R2=0.36; p=0.001). Conclusion: We identified significant differences in microbiota composition between women and between breasts of the same woman. These results warrant further investigation to elucidate the potential relationship between breast tissue microbiota and breast cancer. Citation Format: Emily M. Klann, Jessica M. Williamson, Massimiliano S. Tagliamonte, Maria Ukhanova, Jaya Ruth Asirvatham, Harvey Chim, Lusine Yaghjyan, Volker Mai. Microbiota composition in bilateral healthy breast tissue and breast tumors [abstract]. In: Proceedings of the AACR Special Conference on the Microbiome, Viruses, and Cancer; 2020 Feb 21-24; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2020;80(8 Suppl):Abstract nr B31.

Arsenic, cadmium, chromium and nickel in cancerous and healthy tissues from patients with head and neck cancer