Multidisciplinary approach to Cheetahs affected by gastrointestinal disease: study of the immune profile and of the fecal proteome

“One Health” concept links human health, animal health and the environment with a great potential to mutually influence the health of all species. Infectious diseases associated with wildlife can worse negative public perceptions of wild animals and public support for conservation requiring conscientious effort and urgency to avoid an undesirable outcome 2 . The exponential growth of the human population led to ecologic drastic changes related to human encroachment on wildlife habitats 3 . During the 20th century, growth of human population, invasion of wildlife habitats, changes in agricultural practices, domestication of wild animals and ecotourism have damaged the conservation of wildlife 4,5 . Establishing and maintaining protected areas are key tools for biodiversity conservation. However, this approach is insufficient for wide-ranging species. Protected areas, created for conservation purposes, do not always adequately conserve biodiversity, many terrestrial protected areas within human-dominated systems are isolated from one another increasing the risk of species extinctions 6 . The relationship between isolation and extinction is the basis of the “metapopulation theory” that many spatially distinct subpopulations reconnected by movement of individuals, leading to genetic exchange and the possibility of re- establishing extirpated subpopulations 7 . Numerous emerging infectious diseases have arisen from, or have been identified in, wildlife with implications for human health. For this reason, wildlife conservation can be considered an important resource of protection8 . Since 1960 the IUCN Red List has made multiple lists for a compendium of animals and plants conservation. To date, more than 134000 species have been assessed for The IUCN Red List, of these 26% are mammals 9 . In thisthesis, different aspectsrelated to an animal classified as vulnerable by the IUCN Red List, the cheetah, will be focused. The aim is to investigate the gastrointestinal physiopathology of cheetahs, with a morbidity rate of 95% in captive animals, correlating it with the immune response. Different studies compared diseases in captive and free-ranging cheetahs showing the prevalence of stress-induced pathologies 10. The reduction of the population has led several cheetahs to be housed in zoos, in conditions that do not allow carrying the physiological behaviors of the species 10. For this reason, some pathologies are defined as probably stress-induced. The present research project is based on a multidisciplinary approach to the physiopathology of “cheetahs” GI diseases. In the first part of the study, starting from a common viral disease of cats (feline infectious peritonitis - FIP) in which the immune system plays a key role in disease progression, methods of monocytes’ isolation from peripheral blood were investigated, and then macrophages were studied regarding phagocytosis and respiratory burst activities. The method was firstly performed from samples of cats with feline infectious peritonitis (FIP) and cats’ samples positive for Feline coronavirus but not FIP, and then performing the same evaluations in samples from healthy cheetahs and from subjects with GI disease associated to Helicobacter spp. In the second part of project, the study of the cytokines profile on the same samples was performed, comparing results between animal populations (healthy vs diseased cheetahs). In the third line of research, fecal proteomics to investigate possible differences in healthy and diseased cheetahs was performed

Study of Macrophage Activity in Cats with FIP and Naturally FCoV-Shedding Healthy Cats

Coronavirus frequently infects humans and animals, showing the ability to recombine and cross over to different species. Cats can be considered a model for studying coronavirus infection, in which feline coronavirus (FCoV) represents a major enteric pathogen related to gastroenteric disease. In this animal, the virus can acquire tropism for macrophage cells, leading to a deadly disease called feline infectious peritonitis (FIP). In this study, monocyte-derived macrophages were isolated by CD14-positive selection in venous whole blood from 26 cats with FIP and 32 FCoV-positive healthy cats. Phagocytosis and respiratory burst activities were investigated and compared between the groups. This is the first study comparing macrophage activity in cats affected by FIP and healthy cats positive for FCoV infection. Our results showed that in cats with FIP, the phagocytic and respiratory burst activities were significantly lower. Our results support the possible role of host immunity in Coronaviridae pathogenesis in cats, supporting future research on the immune defense against this systemic disease

Therapeutic approaches to coronavirus infection according to “One Health” concept

Coronaviridae constantly infect human and animals causing respiratory, gastroenteric or systemic diseases. Over time, these viruses have shown a marked ability to mutate, jumping over the human-animal barrier, thus becoming from enzootic to zoonotic. In the last years, numerous therapeutic protocols have been developed, mainly for severe acute respiratory syndromes in humans. The aim of this review is to summarize drugs or other approaches used in coronavirus infections focusing on different roles of these molecules or bacterial products on viral adhesion and replication or in modulating the host's immune system. Within the "One Health" concept, the study of viral pathogenic role and possible therapeutic approaches in both humans and animals is essential to protect public health

Evaluation of the dog population in two Italian shelters in Central Italy (Marche region) as potential blood donors

Background: In recent years, the field of transfusion medicine for dogs has advanced significantly, becoming a crucial aspect of veterinary clinical practice. Nowadays, blood still remains a fundamental biological source and the welfare and health status of eligible species-specific blood donors are essential for veterinary transfusion medicine. Aim: This study focused on evaluating two shelters in Central Italy (Marche region), located in Tolentino (TS) and in Camerino (CS), in order to assess the potential of the shelter dogs as canine blood donors. Methods: We evaluated a total of 45 dogs from these shelters based on physical (age and size), clinical, behavioural, and blood analysis criteria described in the Italian Ministerial Guideline for Veterinary Transfusion Medicine (2016). Results: At the TS shelter, out of 206 resident dogs, 125 met the donation criteria (60.68%), with 28 (13.59%) selected for the study due to the impossibility to collect the samplings or other exclusion causes. In the CS shelter, of the 149 dogs, 17 (11.41%) were identified as potential blood donors and included in the study. Among these, seven dogs (25%) from TS and five dogs (29.41%) from CS were found to have DEA1 negative blood group. High percentages (TS = 25.24%, CS = 40.27%) of dogs were excluded for seniority and 29.53% in CS for behavioural issues. Notable findings included reduced erythrocyte mean corpuscular volume [mean TS = 63.93 fl; CS = 64.00 fl] and Reticulocyte Hemoglobin [mean TS = 22.39 pg; CS = 21.38 pg]. Additionally, in both shelters’ dogs showed a modest increase in eosinophils levels [mean TS = 1.59 K/μl; CS = 1.02 K/μl]. Conclusion: Shelter dogs can fulfill the blood donation requirements set by the Italian Ministerial Guideline on Veterinary Transfusion Medicine. They are generally in good health and present a low risk of transmitting parasitic diseases; however, many are ineligible for donation due to behavioural pathologies or temperament issues and seniority

Canine Blood Group Prevalence and Geographical Distribution around the World: An Updated Systematic Review

“Blood group” or “blood type” refers to the blood group system comprising red blood cell antigens and a specific pattern. Many studies have characterized dog blood groups by the prevalence of the Dog Erythrocyte Antigen (DEA), Kai, and Dal antigens in different geographic areas and by using a variety of methods. Some pioneering studies on blood groups, upon which others were subsequently based, were conducted by Bowdler, Colling, and Hall in the 1970s and 1980s. Our results show that most relevant studies covered the European and American continents, and the methods used to identify DEA, Kai, and Dal groups evolved from 1999 to 2020, albeit without a single method based upon specificity and sensitivity. However, the existence of rapid commercial kits for the determination of the DEA 1 group makes this group the most widely used in clinical practice. Through a systematic review, our aim was to illustrate the countries in the world where different blood groups have been identified with reference to the different methods used and the prevalence of those groups among dog breeds

Updates on Laboratory Evaluation of Feline Cardiac Diseases

Laboratory tests can be altered in cardiovascular diseases and the investigation of specific tests or biomarkers may provide additional information about myocardial damage. Traditional laboratory tests, such as cell blood count, serum biochemistry, and coagulation, can be useful in investigating patients, but are not specific. However, markers like Troponin and Natriuretic Peptides may possibly furnish further data on myocardium damage and can be used in both studying and monitoring cats with cardiac disease. Moreover, the evaluation of the thyroid profile is very important as hyperthyroid cats concomitant cardiovascular diseases are very common and they can also be a direct consequence of endocrinopathy. The purpose of this manuscript is to provide the widest possible overview of what is present in the literature about the feline clinical pathology of heart diseases through a rational division of the main alterations of traditional tests and biomarkers

Evaluation of the Fecal Proteome in Healthy and Diseased Cheetahs (Acinonyx jubatus) Suffering from Gastrointestinal Disorders

Fecal proteomics allows for the identification of proteins and peptides present in stools and is useful in finding possible new biomarkers for diagnosing and/or monitoring gastrointestinal (GI) disorders. In the present study, we investigated the fecal proteome in healthy and diseased cheetahs (Acinonyx jubatus). Captive individuals of this species frequently show gastrointestinal disorders characterized by recurrent episodes of diarrhea, rare episodes of vomiting and weight loss, associated with Helicobacter spp. infection. Fecal proteomic evaluation has been performed by two-dimensional electrophoresis followed by liquid chromatography-tandem mass spectrometry. In healthy cheetahs, the results showed the presence of the following proteins: collagen alpha-1 (II) chain, transthyretin, IgG Fc-binding protein, titin, dystonin, isopentenyl-diphosphate Delta-isomerase 1, sodium/potassium-transporting ATPase subunit alpha-1 and protein disulfide-isomerase A6. The presence of albumin isoforms was found only in diseased cheetahs. The present paper reports the study of the fecal proteome in the cheetah, evidences some differences between healthy and diseased patients and confirms, once again, the potential of fecal proteomics for the study of the GI environment, with promising developments regarding the identification of new diagnostic/monitoring markers

ROLE OF XENOSIALIZATION IN THE PATHOGENESIS OF COLITIS IN THE CANINE MODEL: POSSIBLE POSITIVE EFFECTS OF NEW PROBIOTICS BLENDS

Inflammatory bowel disease (IBD) is a heterogeneous state of chronic intestinal inflammation with no exact known cause. In dogs, IBD is expressed histologically with lymphoplasmacytic inflammation (LPL) of gastrointestinal tract. The study evaluates the possible correlation between microbiota, dietary absorption,enteric expression of N-glycolylneuraminic acid (Neu5Gc) (xenosialization) and intestinal inflammation (xenosialitis) in dogs with IBD. Neu5Gc is indeed synthesized from its N-acetyl precursor (Neu5Ac) by cytidine-5′-monophospho-N acetylneuraminic acid hydroxylase (CMAH), absent in humans, and polymorphic in dogs

The probiotic SLAB51 as agent to counteract BPA toxicity on zebrafish gut microbiota -liver-brain axis

A plethora of studies have so far described the toxic effects of bisphenol A (BPA) on organism health, highlighting the urgent need to find new strategies not only to reduce the presence of this toxicant but also to counteract its adverse effects. In this context, probiotics emerged as a potential tool since they promote organism welfare. Using a multidisciplinary approach, this study explores the effects of SLAB51 dietary administration to counteract BPA toxicity using zebrafish as a model. Adult males and females were maintained under standard conditions (control group; C), exposed for 28 days via the water to an environmental relevant dose of BPA (10 mu g/L; BPA), dietary treated with SLAB51 (109 CFU/g of body weight; P) and co -treated with BPA plus SLAB51 (BPA + P). In the gut, exposure to BPA resulted in altered architecture in both males and females, with females also experiencing an increase of pathogenic bacterial species. Co -administration of BPA + P led to the restoration of normal gut architecture, favored beneficial bacteria colonization, and decreased the abundance of pathogenic species. In the liver, male BPA exposure led to steatosis and glycogen depletion, which was partially mitigated by SLAB51 co -administration. In contrast, in females exposed to BPA, the lack of steatosis along with the greater glycogen depletion, suggested an increase in energy demand as supported by the metabolomic phenotype. The analysis of liver metabolites in BPA + P males revealed increased levels of anserine and reduced levels of glutamine, which could lie behind the counteraction of the brain histopathological damage caused by BPA. In BPA + P females, a reduction of retinoic acid was found in the liver, suggesting an increase in retinoids responsible for BPA detoxification. Overall, these results demonstrate that SLAB51 exerts its beneficial effects on the gut microbiota-brain-liver axis through distinct molecular pathways, effectively mitigating the pleiotropic toxicity of BPA

Faecal proteome in clinically healthy dogs and cats: Findings in pooled faeces from 10 cats and 10 dogs

Background: In the scientific literature, there are only a few manuscripts available on small animal faecal proteomics. Methods: In the present pilot study, this evaluation was performed using pooled faecal samples from 10 clinically healthy dogs and, for the first time, in 10 clinically healthy cats by mean of two-dimensional electrophoresis followed by liquid chromatography-tandem mass spectrometry. Results: Our results showed the presence of nine (albumin, alkaline phosphatase, chymotrypsin-C-like, cytosol aminopeptidase, elastase-3B/proteinase E, immunoglobu-lins and nuclear pore membrane glycoprotein 210) and 14 (albumin, caspase recruitment domain-containing protein, chymotrypsin-like, deleted in malignant brain tumours 1 protein-like, hypothetical protein LOC107375, immunoglobulin, kallikrein-1, superox-ide dismutase, transthyretin precursor, interstitial collagenase-like) different proteins in canine and feline faeces, respectively. Conclusion: These preliminary findings document the presence of a range of proteins in the faeces of apparently healthy dogs and cats and may serve as a basis for larger, prospective studies to establish reference proteomic data against which diseased popu-lations can be compared