Proteomic signature profiling in the cortex of dairy cattle unravels the physiology of brain aging.

Introduction: Aging is a physiological process occurring in all living organisms. It is characterized by a progressive deterioration of the physiological and cognitive functions of the organism, accompanied by a gradual impairment of mechanisms involved in the regulation of tissue and organ homeostasis, thus exacerbating the risk of developing pathologies, including cancer and neurodegenerative disorders. Methods: In the present work, for the first time, the influence of aging has been investigated in the brain cortex of the Podolica cattle breed, through LC-MS/MS-based differential proteomics and the bioinformatic analysis approach (data are available via ProteomeXchange with identifier PXD044108), with the aim of identifying potential aging or longevity markers, also associated with a specific lifestyle. Results and discussion: We found a significant down-regulation of proteins involved in cellular respiration, dendric spine development, synaptic vesicle transport, and myelination. On the other hand, together with a reduction of the neurofilament light chain, we observed an up-regulation of both GFAP and vimentin in the aged samples. In conclusion, our data pave the way for a better understanding of molecular mechanisms underlying brain aging in grazing cattle, which could allow strategies to be developed that are aimed at improving animal welfare and husbandry practices of dairy cattle from intensive livestock.

Preliminary evaluation of the proteomic profiling in the hippocampus of aged grazing cattle.

Brain aging is a physiological process associated with physical and cognitive decline; however, in both humans and animals, it can be regarded as a risk factor for neurodegenerative disorders, such as Alzheimer's disease. Among several brain regions, hippocampus appears to be more susceptible to detrimental effects of aging. Hippocampus belongs to limbic system and is mainly involved in declarative memories and context-dependent spatial-learning, whose integrity is compromised in an age-dependent manner. In the present work, taking advantage of liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based proteomics, we sought to identify proteins differentially expressed in the hippocampus of the aged grazing milk cows. Our exploratory findings showed that, out of 707 identified proteins, 112 were significantly altered in old cattle, when compared to the adult controls, and functional clusterization highlighted their involvement in myelination, synaptic vesicle, metabolism, and calcium-related biological pathways. Overall, our preliminary data pave the way for the future studies, aimed at better characterizing the role of such a subcortical brain region in the age-dependent cognitive decline, as well as identifying early aging markers to improve animal welfare and husbandry practices of dairy cattle from intensive livestock.

Description of Twenty-Nine Animal Hoarding Cases in Italy: The Impact on Animal Welfare.

The hoarding of animals is a psychiatric disease, characterized by a compulsive collection of animals, with a relevant impact upon the care and welfare of animals, as well as on human society. In Italy, there are neither substantial reports nor information shared about such a phenomenon, making it difficult to draw a clear picture of the hoarder profile. Therefore, in the present work, we sought to detail 29 cases of animal accumulators in Italy, who lived within two areas of the Lazio region, and accumulated a total of 1080 animals from 2019 to 2022. In line with other international studies, we observed a prevalence of middle-aged (in their fifties) women, who lived mainly alone in a high level of social and health degradation. Most of the hoarded animals exhibited severe signs of dehydration and malnutrition, muscle hypotrophy, dermatological injuries, and behavioral disorders. Animal hoarding is not yet fully understood nor recognized as a psychosocial disorder, although it produces a deep suffering for the hoarder themselves, as well as corresponding family members, and the animals accumulated. Therefore, given the crucial impact of animal hoarding upon human and animal welfare, cross-cultural networks aimed at properly raising awareness of the problem could be established.

Combining α-s1 Casozepine and Fluoxetine Treatment with a Behavioral Therapy Improves Symptoms in an Aggressive Dog: An Italian Case Report.

Behavioral dysfunctions in dogs represent a critical issue of the human-animal relationship. In particular, aggression can make interspecific coexistence quite complicated within family units, thus exposing all members to greater health risks. In this present study, we documented multiple aggression episodes against one of the two family members caused by a 4-year-old neutered male pure breed Lagotto Romagnolo dog. To minimize impulsivity and anxiety-like behaviors of the patient as much as possible and improve his relationship with the adopting family, we used an interdisciplinary approach, employing specific skilled personnel, including a veterinary behaviorist and a rehabilitating dog instructor. Nine months after fluoxetine treatment (0.8 mg/kg, SID), in combination with oral α-s1 casozepine administration, and behavioral rehabilitation, the owners reported a significant reduction in aggressive events in terms of intensity and frequency. Collectively, our promising data pave the way toward a more detailed characterization of α-s1-casozepine to better evaluate the potential involvement of such a compound in the modulation of aggressive behaviors in dogs affected by relational dysfunctions.

Combining cannabis and melatonin treatment with a rehabilitation program improved symptoms in a dog with compulsive disorder: A case report.

Compulsive disorder in dogs (CD) is characterized by constant and time-consuming repetition of behaviors, emancipated from the environment, that definitely compromise their everyday life activities. Here, we documented the efficacy of a novel approach to counteract the negative symptoms of CD in a 5-year-old mongrel dog, previously found to be resistant to the conventional antidepressant. The patient underwent an integrated and interdisciplinary approach, based on the cannabis and melatonin co-administration, together with a tailored 5-month-lasting behavioral program. Observational findings showed a lower rate of compulsive episodes and better management of the dog as well, when compared to the previous paroxetine treatment. We followed him for an additional four months of therapy, and the owners reported easier management of the dog, as reduction of abnormal behaviors to a level acceptable to the owners. Overall, our data so far collected in the CD dog may allow us to test more deeply the feasibility and safety of such an off-label approach, at both preclinical and clinical levels.

Puppies Raised during the COVID-19 Lockdown Showed Fearful and Aggressive Behaviors in Adulthood: An Italian Survey.

During the COVID-19 pandemic, the Prime Minister's decree issued by the Italian government on 9 March 2020, known as "#Iorestoacasa" (I stay at home), required everyone to stay indoors, with a few exceptions, from 11 March to 3 May 2020. This decree had a substantial impact on the mental health of both people and dogs. We carried out a national survey to compare the personalities of adult dogs who were puppies in lockdown (from March to May 2020) with those of adult dogs born after that time (from June 2020 to February 2021). Our results highlighted a significant increase in personality traits related to fear and aggression in dogs who experienced lockdown restrictions during their socialization period, further confirming that the COVID-19 pandemic strongly impacted the behavioral development of dogs. Therefore, it may be advantageous for these dogs to be closely monitored by veterinary behaviorists and receive specialized behavioral rehabilitation therapy to lower the risk of episodes of aggression and fearfulness as well as to increase the wellbeing of dogs raised under social restrictions.

Relationship among Milk Conductivity, Production Traits, and Somatic Cell Score in the Italian Mediterranean Buffalo.

The measurement of milk electrical conductivity (EC) is a relatively simple and inexpensive technique that has been evaluated as a routine method for the diagnosis of mastitis in dairy farms. The aim of this study was to obtain further knowledge on relationships between EC, production traits and somatic cell count (SCC) in Italian Mediterranean Buffalo. The original dataset included 5411 records collected from 808 buffalo cows. Two mixed models were used to evaluate both the effect of EC on MY, PP and FP and EC at test-day, and the effect of EC on somatic cell score (SCS) by using five different parameters (EC_param), namely: EC collected at the official milk recording test day (EC_day0), EC collected 3 days before official milk recording (EC_day3), and three statistics calculated from EC collected 1, 3 and 5 days before each test-day, respectively. All effects included in the model were significant for all traits, with the only exception of the effect of EC nested within parity for FP. The relationship between EC and SCS was always positive, but of different magnitude according to the parity. The regression of EC on SCS at test-day using different EC parameters was always significant except when the regression parameter was the slope obtained from a linear regression of EC collected over the 5-day period. Moreover, in order to evaluate how well the different models fit the data, three parameters were used: the Average Information Criteria (AIC), the marginal R2 and the conditional R2. According to AIC and to both the Marginal and Conditional R2, the best results were obtained when the regression parameter was the mean EC estimated over the 5-day period.

Immunolocalization of Nesfatin-1 in the Gastrointestinal Tract of the Common Bottlenose Dolphin Tursiops truncatus.

First identified as an anorexigenic peptide, in the last decades, several studies have suggested that Nesfatin-1 (Nesf-1) is a pleiotropic hormone implicated in numerous regulatory processes in peripheral organs and tissues. In vertebrates, Nesf-1 is indeed expressed in the central nervous system and peripheral organs. In this study, we characterized the pattern of Nesf-1 distribution within the digestive tract of the common bottlenose dolphin (Tursiops truncatus), composed of three gastric chambers and an intestine without a clear subdivision in the small and large intestine, also lacking a caecum. Our results indicated that Nesf-1 is widely distributed in cells of the mucosal epithelium of the gastric chambers. Most of the immunoreactivity was observed in the second chamber, compared to the first and third chambers. Immunopositivity was also found in nerve fibers and neurons, scattered or/and clustered in ganglion structures along all the examined gastrointestinal tracts. These observations add new data on the highly conserved role of Nesf-1 in the mammalian digestive system.

Serological Evidence of Q Fever among Dairy Cattle and Buffalo Populations in the Campania Region, Italy.

Due to its economic impact on livestock and its zoonotic effect, Q fever is a public and animal health problem. Information on this infection in Italy is presently supported by reports of reproductive problems in livestock farms and is, therefore, insufficient to properly understand the impact of the disease. This study aimed to describe for the first time the seroprevalence of Q fever in dairy cows and water buffalos in the Campania region (Southern Italy). A total of 424 dairy cattle and 214 water buffalo were tested using a commercial indirect ELISA kit. An overall seroprevalence of 11.7% confirmed the wide distribution of C. burnetii in this region. Several factors were positively associated with higher seroprevalence, such as species (higher in cattle than in water buffalo), age, and coexistence with other ruminant species. The final model of logistic regression included only age (older) and species (cattle), which were positively associated with the presence of Q fever antibodies. Our findings support the widespread presence of Coxiella burnettii in Campania and show a seroprevalence similar to that observed in previous studies in other Italian regions and European countries. Since human cases are typically linked to contact with infected ruminants, there is a need to improve surveillance for this infection.

The Key Role of Lysosomal Protease Cathepsins in Viral Infections.

Cathepsins encompass a family of lysosomal proteases that mediate protein degradation and turnover. Although mainly localized in the endolysosomal compartment, cathepsins are also found in the cytoplasm, nucleus, and extracellular space, where they are involved in cell signaling, extracellular matrix assembly/disassembly, and protein processing and trafficking through the plasma and nuclear membrane and between intracellular organelles. Ubiquitously expressed in the body, cathepsins play regulatory roles in a wide range of physiological processes including coagulation, hormone secretion, immune responses, and others. A dysregulation of cathepsin expression and/or activity has been associated with many human diseases, including cancer, diabetes, obesity, cardiovascular and inflammatory diseases, kidney dysfunctions, and neurodegenerative disorders, as well as infectious diseases. In viral infections, cathepsins may promote (1) activation of the viral attachment glycoproteins and entry of the virus into target cells; (2) antigen processing and presentation, enabling the virus to replicate in infected cells; (3) up-regulation and processing of heparanase that facilitates the release of viral progeny and the spread of infection; and (4) activation of cell death that may either favor viral clearance or assist viral propagation. In this review, we report the most relevant findings on the molecular mechanisms underlying cathepsin involvement in viral infection physiopathology, and we discuss the potential of cathepsin inhibitors for therapeutical applications in viral infectious diseases.

The Potential Impact of a Dog Training Program on the Animal Adoptions in an Italian Shelter.

One of the main concerns of the human-dog relationship is today associated with the quality life inside the kennels, which are very often regarded as animal dump where dogs are exiled, representing a burden on society. In the present study we sought to investigate the importance of performing an appropriate behavioral program on the adoption chances within an Italian shelter, near Naples (Ottaviano). In this respect, we enrolled 555 adopted dogs of different ages, who followed a tailored-4-month lasting training program between 2018 and 2020. Once entered there, they were carefully examined by the veterinary behaviorist, and directed towards a suited training program, to improve living conditions. We documented a higher number of both adult and senior dogs who left the kennel and were adopted, compared to the age-matched untrained animals (n = 479), housed in the same kennel from 2015 to 2017. Taken together, the present data highlight an important role for training in improving the natural attitudes of the companion dogs, thus pointing towards a better human-animal bond.

An Interdisciplinary Approach for Compulsive Behavior in Dogs: A Case Report.

Compulsive disorder is a debilitating condition affecting both humans and animals, characterized by intrusive thoughts and recurring out-of-place behaviors. Among them, tail chasing might represent one of the most common traits in compulsive dogs. Herein, we reported the case of a 7-year-old intact male German Shepherd mixed-breed dog, presenting with tail chasing behavior. He underwent a first behavioral evaluation 1 year before (at the age of 6), when he injured himself with severe wounds at level of the tail and left thigh. To avoid any specific suffering and increase his physical health, of course, the study was carried out through an interdisciplinary approach, employing a veterinary behaviorist and a rehabilitating dog instructor. Three months after pharmacological treatment with fluoxetine and α-s1 casozepine, associated with a behavioral recovery program, the owner reported an improvement of compulsive events in his dog, in terms of intensity and frequency. Interestingly, over the following 3 months, the dog did not experience any new tail chasing episodes.

Retrospective analysis of dog bites in Southern Italy / Análise retrospectiva de mordidas de cães no sul da Itália

Dog bite is one of the major public health problems involving people worldwide. Although, several studies have investigated this phenomenon in different countries, little information about the incidence of dog bite episodes in Italy is available. We analysed data about dog biting events between 2010 and 2019 provided by the CRIUV, the Regional Reference Centre for Veterinary Urban Hygiene in the largest city of Southern Italy, namely. Naples. We observed severe and profound inconsistencies in the data collection that reveal structural and significant weaknesses of the current data collection system. Given the multifactorial nature of dog bite, we highlight the need to improve the gathering of all the information related to the factors affecting the occurrence of biting episodes for an accurate assessment of the biting phenomenon.

A mordida de cães é um dos maiores problemas de saúde pública que envolve pessoas em todo o mundo. Embora vários estudos tenham investigado esse fenômeno em diferentes países, poucas informações sobre a incidência de episódios de mordidas por cães na Itália estão disponíveis. Analisamos dados sobre eventos de mordidas de cães entre 2010 e 2019 fornecidos pelo CRIUV, Centro de Referência Regional para Higiene Veterinária Urbana na maior cidade do sul da Itália, Nápoles. Observamos inconsistências graves e profundas na coleta de dados que revelam fragilidades estruturais e significativas do sistema de coleta de dados atual. Dada a natureza multifatorial da mordida canina, destaca-se a necessidade de melhorar a coleta de todas as informações relacionadas aos fatores que afetam a ocorrência de episódios de mordida para uma avaliação precisa deste fenômeno.

Human-Dog Relationship during the First COVID-19 Lockdown in Italy.

The SARS-CoV2 pandemic forced an abrupt interruption of social contacts and interpersonal affective relationships all over the world, according to national directives. Many considerable inconveniences occurred with important repercussions also on the emotional state of people and their pets. We carried out a national survey to evaluate the human-dog relationship in a social isolation context using an adapted version of Monash Dog Owner Relationship Scale, the perception of the dogs' discomfort by their human owners, and the resilience of the dog through the quantification of symptoms, in time of the first lockdown of the COVID-19 pandemic. The results highlighted that the human-dog interaction was similar during quarantine; however, there was lower owner's perception of a dog's cost during the quarantine than before it.

Antioxidants in assisted reproductive technologies: An overview on dog, cat, and horse.

Assisted reproductive technologies (ARTs) are widely used as a tool to improve reproductive performance in both humans and animals. In particular, in the veterinary field, ARTs are used to improve animal genetics, recover endangered animals, and produce offspring in the event of subfertility or infertility in males or females. However, the use of ARTs did not improve the fertilization rate in some animals due to various factors such as the difficulty in reproducing an anatomical and humoral substrate typical of the natural condition or due to the increase in catabolites and their difficult elimination. The in vitro environment allows the production and increase in the concentration of substances, including reactive oxygen species (ROS), which could be harmful to gametes. If produced in high concentration, the ROS becomes deleterious, both in vitro and in vivo systems. It has been seen that the use of antioxidants can help neutralize or counteract the production of ROS. The present study aims to report the latest findings regarding the use of antioxidants in ARTs of some domestic species, such as dogs, cats, and horses, compared to other animal species, such as cattle, in which ARTs have instead developed more widely.

Cortisol Levels of Shelter Dogs in Animal Assisted Interventions in a Prison: An Exploratory Study.

Previous studies regarding the Animal Assisted Interventions (AAI) have mainly focused on the beneficial effects of human-animal interactions on human health; whereas the impact of such activities on the welfare of the animals involved has received limited attention. So far, few studies have addressed this issue by evaluating the physiological and behavioral reactions of therapy dogs during the interventions. The aim of this study was to evaluate the potential effect of AAI on the cortisol levels of shelter dogs. Five dogs participated in weekly AAI working activities with adult inmates held at a prison of the South of Italy for two months. Saliva samples were collected every two weeks in three conditions: at the kennel (baseline), after transportation and at the end of the working sessions. The results revealed a significant decrease in the cortisol baseline at the end of the AAI program, suggesting that the activities carried out with humans and in a different environment could improve the welfare of dogs housed in kennels. Moreover, we found that transportation significantly increased subjects' cortisol levels, suggesting that it is a critical phase that deserves particular care.

Do Animals Play a Role in the Transmission of Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2)? A Commentary.

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) belongs to the Beta-coronavirus genus. It is 96.2% homologous to bat CoV RaTG13 and 88% homologous to two bat SARS-like coronaviruses. SARS-CoV-2 is the infectious agent responsible for the coronavirus disease (COVID-19), which was first reported in the Hubei province of Wuhan, China, at the beginning of December 2019. Human transmission from COVID-19 patients or incubation carriers occurs via coughing, sneezing, speaking, discharge from the nose, or fecal contamination. Various strains of the virus have been reported around the world, with different virulence and behavior. In addition, SARS-CoV-2 shares certain epitopes with some taxonomically related viruses, with tropism for the most common synanthropic animals. By elucidating the immunological properties of the circulating SARS-CoV-2, a partial protection due to human-animal interactions could be supposed in some situations. In addition, differential epitopes could be used for the differential diagnosis of SARS-CoV-2 infection. There have been cases of transmission from people with COVID-19 to pets such as cats and dogs. In addition, wild felines were infected. All These animals were either asymptomatic or mildly symptomatic and recovered spontaneously. Experimental studies showed cats and ferrets to be more susceptible to COVID-19. COVID-19 positive dogs and felines do not transmit the infection to humans. In contrast, minks at farms were severely infected from people with COVID-19. A SARS-Cov-2 variant in the Danish farmed mink that had been previously infected by COVID-19 positive workers, spread to mink workers causing the first case of animal-to-human infection transmission that causes a moderate decreased sensitivity to neutralizing antibodies. Thus, more investigations are necessary. It remains important to understand the risk that people with COVID-19 pose to their pets, as well as wild or farm animals so effective recommendations and risk management measures against COVID-19 can be made. A One Health unit that facilitates collaboration between public health and veterinary services is recommended.

Triple-Negative Breast Cancer Comparison With Canine Mammary Tumors From Light Microscopy to Molecular Pathology.

Many similar characteristics in human and dog cancers including, spontaneous development, clinical presentation, tumor heterogeneity, disease progression, and response to standard therapies have promoted the approval of this comparative model as an alternative to mice. Breast cancer represents the second most frequent neoplasm in humans after lung cancer. Triple-negative breast cancers (TNBC) constitute around 15% of all cases of breast cancer and do not express estrogen receptor (ER), progesterone receptor (PR), and do not overexpress human epidermal growth factor receptor 2 (HER2). As a result, they do not benefit from hormonal or trastuzumab-based therapy. Patients with TNBC have worse overall survival than patients with non-TNBC. Lehmann and collaborators described six different molecular subtypes of TNBC which further demonstrated its transcriptional heterogeneity. This six TNBC subtype classification has therapeutic implications. Breast cancer is the second most frequent neoplasm in sexually intact female dogs after skin cancer. Canine mammary tumors are a naturally occurring heterogeneous group of cancers that have several features in common with human breast cancer (HBC). These similarities include etiology, signaling pathway activation, and histological classification. Molecularly CMTs are more like TNBCs, and therefore dogs are powerful spontaneous models of cancer to test new therapeutic approaches, particularly for human TNBCs. More malignant tumors of the breast are more often ER and PR negative in both humans and dogs. Promising breast cancer biomarkers in both humans and canines are cancer-associated stroma (CAS), circulating tumor cells and tumor DNA (ctDNA), exosomes and miRNAs, and metabolites.

Human-Animal Relationship Dysfunction: A Case Study of Animal Hoarding in Italy.

"Animal hoarding" or "compulsive hoarding of animals" is a psychiatric disease, which has important social implications and a profound influence on animal welfare. To date, this phenomenon has been little investigated and largely unexplored. The present study aims to systematically describe a case of animal hoarding, which remains unresolved. The report refers to a case of a woman suffering from animal hoarding that emerged in 2005. From March 2014 to December 2019, 450 animals were seized over nine different occasions. This disease had significant implications on the welfare of the animals collected, which lived in poor housing and hygiene conditions that frequently led to their death. Since animal hoarding cases involve sanitary, legal, and veterinary aspects, we believe that a multidisciplinary approach is necessary in order to prevent a recurrence and a new accumulation of animals. A holistic approach should be taken according to the One Health principle that involves different stakeholders at every level in order to adopt an efficient solution.

Geographical information system analysis on road accidents involving wandering dogs in the urban area of Naples.

In this study, the spatial distribution of road accidents involving wandering dogs on urban roads of Naples was assessed using kernel density estimation (KDE). The study included 423 dogs victim of road collisions in the period 2012-2015 collected from the medical records of Interdepartmental Centre of Veterinary Radiology, of Pathological Anatomy Service, of Veterinary Teaching Hospital, "Federico II" University of Naples, and of Veterinary Hospital Attendance of the Local Public Health Unit Napoli1. There were a significant prevalence of young male dogs and the KDE demonstrated the presence of five hotspots in five city neighbourhoods. All areas with the highest KDE were found to be peripheral zones in which wide green areas, uncultivated or cultivated, merged with built areas and crossed by large straight roads with a lot of intersections. Wandering dogs as other animals represent a mounting problem on urban roads in Italy, despite of the effort to reduce these populations. Better knowledge of this phenomenon is important with reference to taking countermeasures and improve road safety. It is also important with reference to One Health and the Decade of Action for Road Safety 2011- 2020 of the United Nations.