Animal Pathology Research Articles

Single image super-resolution for whole slide image using convolutional neural networks and self-supervised color normalization.

High-quality whole slide scanners used for animal and human pathology scanning are expensive and can produce massive datasets, which limits the access to and adoption of this technique. As a potential solution to these challenges, we present a deep learning-based approach making use of single image super-resolution (SISR) to reconstruct high-resolution histology images from low-resolution inputs. Such low-resolution images can easily be shared, require less storage, and can be acquired quickly using widely available low-cost slide scanners. The network consists of multi-scale fully convolutional networks capable of capturing hierarchical features. Conditional generative adversarial loss is incorporated to penalize blurriness in the output images. The network is trained using a progressive strategy where the scaling factor is sampled from a normal distribution with an increasing mean. The results are evaluated with quantitative metrics and are used in a clinical histopathology diagnosis procedure which shows that the SISR framework can be used to reconstruct high-resolution images with clinical level quality. We further propose a self-supervised color normalization method that can remove staining variation artifacts. Quantitative evaluations show that the SISR framework can generalize well on unseen data collected from other patient tissue cohorts by incorporating the color normalization method.

Incoherent polarized light in treatment of purulent wounds in pigs

The treatment of wounds in surgery is one of the important problems that must be solved taking into account animal species, its general condition, the stage of the wound process, and also the type of wound. The aim of our research was to study the effectiveness of incoherent polarized light, the source of which is the Bioptron lamp, in combination with other medicines, in the treatment of purulent wounds in pigs and to monitor changes in their blood composition. The studies were carried out on large white pigs who were diagnosed with occasional purulent wounds of soft tissues in different parts of the body in the conditions of LLC “SPhU AGRO” in the village of Grozino, Korosten district of Zhytomyr region. A total of 675 pigs were examined .There were found 11 animals with purulent wounds out of which an experimental and control groups were formed and appropriate treatment regimens were prescribed. It was found that after applying the treatment regimen, which consisted of the use of incoherent polarized light, the source of which is the “Bioptron” lamp, gauze drainage with methyrulacil ointment and Nitacid ointment, as well as intramuscular injections of amoxicillin, the purulent wounds in the experimental pigs completely healed on average by Day 16-17, and in the control, where the traditional method of treatment was used (surgical treatment of the wound, drainage from a 3% of boric acid, 10 % Catosal, 5 % synthomycin emulsion; muscularly 15 % amoksikel), complete healing was observed on the 19-20th day. We attribute more rapid wound healing in observed animals with wound healing activity of Nitacid ointment in combination with irradiation by incoherent polarized light. Changes were found in pigs' blood tests on purulent wounds, in comparison with healthy animals, except: decrease in hemoglobin, erythropenia leukocytosis, significant increase in erythrocyte sedimentation and also decease in biochemical idicators. The use of medicinal treatment and physiotherapeutic procedures are needed to shorten the healing of purulent wounds in pigs. Further research will focus on the use of incoherent polarized light, radiated by the “Bioptron” lamp for the surgical pathology of animals. Key words: incoherent polarized light, “Bioptron” lamp, purulent wounds, pigs, wounds healing, blood.

Hematological indexes of dogs at dirofilariosis

Laboratory diagnostics for any animal pathology refers to special research methods and helps not only in establishing the final diagnosis, but also in expanding data on the course of primary pathology and determining secondary complications based on changes in blood parameters. For this purpose, a clinical study of the blood of dogs with varying degrees of intensity of dirofilariasis invasion was carried out. It was found that the number of erythrocytes decreased in animals of the experimental groups, depending on the intensity of invasion. Severe erythropenia was found in the group of animals with the highest degree of invasion intensity (2.04±0.50 1012/l (p<0.001). It increased with an increasing in the degree of invasion and ESR up to 40.28±12.67 mm/h (p<0.05) and 62.50±14.16 mm/h (p<0.01). Thrombocyto-penia at 199.13±55.31 109/l was found in dogs in the group with the highest degree of invasion. Oligochromemia in animals also depended on the intensity of invasion and was 94.44±12.30 g/l (p<0.05) and 52.00±12.69 g/l (p<0.001). In dogs with the highest number of dirofilaria larvae, the hematocrit was 19.75±6.34 % (p<0.01) and leukocytosis developed (20.53±8.91 109/l). By calculating the leukocyte count in dogs with the highest degree of dirofilariasis invasion, a significant increase in the number of segmented neu-trophils was found to 86.00±10.43 % (p<0.1), eosinophils up to 27.88±5.54 % (p<0.001), basophils up to 1.75±1.39 % (p<0.001), appeared in blood of sick animals young neutro-phils (0.50±1.07 %, p<0.001). The established changes in hematological parameters are associated with the development of hemolytic anemia during invasion and the presence of latent bleeding associated with the destruction of the vascular wall by dirofilariae. The relative leukocytosis, which were diagnosed in sick dogs, is associated with the presence of nematodes and the development of inflammatory processes at the sites of localization and migration of dirofilaria. It is possible that leukocytosis and a shift of the leukocyte formula to the left are associated with intoxication of the body. Key words: dogs, heartworm disease, Dirofilaria im-mitis, blood, erythrocytes, leukocytes, leukocyte formula, hemoglobin, hematocrit, erythrocyte sedimentation rate.

Recurrent limbic seizures do not cause hippocampal neuronal loss: A prolonged laboratory study

PurposeIt remains controversial whether neuronal damage and synaptic reorganization found in some forms of epilepsy are the result of an initial injury and potentially contributory to the epileptic condition or are the cumulative affect of repeated seizures. A number of reports of human and animal pathology suggest that at least some neuronal loss precedes the onset of seizures, but there is debate over whether there is further damage over time from intermittent seizures. In support of this latter hypothesis are MRI studies in people that show reduced hippocampal volumes and cortical thickness with longer durations of the disease. In this study we addressed the question of neuronal loss from intermittent seizures using kindled rats (no initial injury) and rats with limbic epilepsy (initial injury). MethodsSupragranular mossy fiber sprouting, hippocampal neuronal densities, and subfield area measurements were determined in rats with chronic limbic epilepsy (CLE) that developed following an episode of limbic status epilepticus (n = 25), in kindled rats (n = 15), and in age matched controls (n = 20). To determine whether age or seizure frequency played a role in the changes, CLE and kindled rats were further classified by seizure frequency (low/high) and the duration of the seizure disorder (young/old). ResultsOverall there was no evidence for progressive neuronal loss from recurrent seizures. Compared with control and kindled rats, CLE animals showed increased mossy fiber sprouting, decreased neuronal numbers in multiple regions and regional atrophy. In CLE, but not kindled rats: 1) Higher seizure frequency was associated with greater mossy fiber sprouting and granule cell dispersion; and 2) greater age with seizures was associated with decreased hilar densities, and increased hilar areas. There was no evidence for progressive neuronal loss, even with more than 1000 seizures. ConclusionThese findings suggest that the neuronal loss associated with limbic epilepsy precedes the onset of the seizures and is not a consequence of recurrent seizures. However, intermittent seizures do cause other structural changes in the brain, the functional consequences of which are unclear.

The effectiveness of “Iodozol” in combination therapy in the treatment of postpartum vulvitis in goats

In the nosology of obstetric pathology of goats a special place is occupied by postpartum lesions of the genitals by pathogenic microflora, of which the lion's share are postpartum vulvitis. The aim of the study was to test a new treatment regimen for postpartum vulvitis in goats and to study the therapeutic efficacy of the drug “Iodozol”, “Celexib” and “Dextrodev 50” in this pathology. Clinical and anamnestic data showed that the symptoms of postpartum vulvitis of goats were characterized by inflammatory edema, high pain, local fever, impaired tissue integrity, hematomas and discharge from the genital tract. Microbiological studies of secretions from the genitals revealed the presence of E. coli, S. aureus, less often – S. saprophyticus and E. faecalis. High resistance of microorganisms to fluoroquinolones and penicillins is noted. E. coli was sensitive to only 35.7 % of antimicrobial drugs, including ceftriaxone, ceftiofur, and iodine. S. aureus is sensitive to 49.9 % of antimicrobial drugs, including ceftriaxone, cephalexin, gentamicin, ceftiofur, iodine (highest sensitivity) and chlorhexidine. S. saprophyticus was insensitive to a number of antibiotics (oxytetracycline, gentamicin, etc.), except iodine. Taking into account microbiological researches the complex scheme of treatment of postpartum vulvitis at goats is developed. Evaluation of the effectiveness of complex therapy of this pathology in animals showed that treatment with “Iodozole”, “Celexib” and “Dextrodev 50” in a short time eliminates inflammatory symptoms (pain, infiltration, redness, discharge). At the end of the course of treatment in the blood was observed an increase in hemoglobin concentration by 13.7 %, a decrease in the number of erythrocytes by 1.7 and leukocytes by 2.2 times compared with treatment. The topical intrauterine drug “Iodozol”, a non-steroidal injectable drug “Celexib”, has been found to have a broad spectrum of antimicrobial activity, and is effective and safe to use. The drugs have an anti-edematous and analgesic effect, and symptomatic treatment with intravenous solution “Dextrodev 50” is a mandatory component of complex therapy and is aimed at replenishing the body with nutrients, as well as helps to neutralize and remove toxins from the body. Further research will be to study the clinical efficacy and develop regulations for the use of this scheme in vulvitis and cervicitis of other ruminants.

Metachromatic Leukodystrophy: Diagnosis, Modeling, and Treatment Approaches.

Metachromatic leukodystrophy is a lysosomal storage disease, which is characterized by damage of the myelin sheath that covers most of nerve fibers of the central and peripheral nervous systems. The disease occurs due to a deficiency of the lysosomal enzyme arylsulfatase A (ARSA) or its sphingolipid activator protein B (SapB) and it clinically manifests as progressive motor and cognitive deficiency. ARSA and SapB protein deficiency are caused by mutations in the ARSA and PSAP genes, respectively. The severity of clinical course in metachromatic leukodystrophy is determined by the residual ARSA activity, depending on the type of mutation. Currently, there is no effective treatment for this disease. Clinical cases of bone marrow or cord blood transplantation have been reported, however the therapeutic effectiveness of these methods remains insufficient to prevent aggravation of neurological disorders. Encouraging results have been obtained using gene therapy for delivering the wild-type ARSA gene using vectors based on various serotypes of adeno-associated viruses, as well as using mesenchymal stem cells and combined gene-cell therapy. This review discusses therapeutic strategies for the treatment of metachromatic leukodystrophy, as well as diagnostic methods and modeling of this pathology in animals to evaluate the effectiveness of new therapies.

Simulation of Acute Destructive Pancreatitis with Damage to the Adjacent Adipose Tissue in Pigs

The aim of the study was to develop a model of acute destructive pancreatitis with damage to the extra-organ adipose tissue. Materials and methods. Simulation of acute destructive pancreatitis with damage of the extra-organ adipose tissue was performed in 22 white breed boars aged 3 months. The animals were divided into groups in accordance with two series of studies, where the control (first) group included 15 animals with the pancreas and extra-organ adipose tissue remained intact. Acute destructive pancreatitis was simulated in 7 pigs of the experimental (second) group based on the proposed technique (patent for an invention No. 2668201). Ultrasound examination of the abdominal cavity and retroperitoneal region was applied to confirm the development of expected pathology in experimental animals. Intensity of blood supply was assessed using contrast radiography by injecting a contrast medium into the superior mesenteric artery. Complete blood count and biochemical assay were performed. In addition, during autopsy tissue samples were taken for histological and immunohistochemical investigations to assess changes in the internal organs. Results. Acute destructive pancreatitis with damage to the extra-organic adipose tissue was regarded as achieved when clinical manifestations of apathy, lethargy, feed refusal were registered in animals. Biochemical blood tests revealed a significant increase in the concentration of alkaline phosphatase and alpha-amylase enzymes. Nitrogenous toxins, in particular, creatinine, were accumulated in the body of the experimental animals. Complete blood count revealed leukocytosis, the fact supporting the onset of the expected inflammatory process in the pancreas and adjacent adipose tissue. The development of necrotic and inflammatory processes in the organ was evidenced by a sharp increase in the level of amylase, creatinine and alkaline phosphatase in the blood of animals. Ultrasound examination revealed a picture of the blurring of pancreatic contours, heterogeneity of the organ structure, increased echogenicity, and enlargement of the duct. Intensification of the vascular pattern and increase in pancreatic contours were detected during the contrast examination of the abdominal vascular system. At autopsy, there were signs of enzymatic peritonitis, namely, serous-hemorrhagic exudate in the abdominal cavity and the cavity of the omental bursa, increased pneumatization of the intestinal tube with a thinning of its wall and foci of small hemorrhages. The pancreatic tissue in all cases was edematous, full-blooded, with multiple foci of fatty necrosis and purulent inflammation. Similar changes were found in the adjacent adipose tissue. At the light-optical level, vascular congestion, small foci of steatonecrosis and micro-abscesses were determined in the pancreas. Immunohistochemical studies largely evidenced the onset of acute destructive pancreatitis with damage to the extra-organ adipose tissue.Conclusion. Thus, the proposed technique allows simulating acute destructive pancreatitis with damage to the extra-organ adipose tissue in pigs, the event frequently occurring in the clinical practice. In animals the disease is accompanied by clear clinical manifestations correlating with findings of instrumental and laboratory investigations, and autopsy results. Investigations of the pancreatic and extra-organ adipose tissue samples at the light-optical and molecular levels evidence the fact that the authors have simulated a model of acute destructive pancreatitis with involvement of the adipose tissue adjacent to the organ in the pathological process.

PROTECTIVE EFFECT OF SOME SALTS 2-AMINOETHANESULFONIC ACID IN AN EXPERIMENTAL MODEL OF DEGENERATIVE SPINAL CORD INJURY

In this work, we studied the pharmacological activity of zinc and magnesium salts of 2-aminoethanesulfonic acid in white non-linear male rats with amyotrophic lateral sclerosis, which was modeled by neurotoxicantsimplication into the pelvic part of spinal cord. After the reproduction of the pathology in animals, the indices of motor activity were recorded in the Rotarod test, and morphological studies of spinal cord sections stained according to Nisl in the Belshovsky modification were carried out. It was shown that the magnesium salt of 2-aminoethanesulfonic acid (compound LHT-317) to a greater extent reduces the development of motor disorders in experimental animals compared with the control group on the 4th day of observation. The course of intravenous administration of the studied compounds of 2-aminoethanesulfonic acid did not inhibit morphological changes in the spinal cord that develop in degenerative-dystrophic pathology of the central nervous system: connections. Moreover, if, against the background of treatment with zinc salt, the total area of motor zones in animals of the experimental group exceeded that of control rats, then the number of motoneurons did not differ from the control.

Loss of TMEM106B potentiates lysosomal and FTLD-like pathology in progranulin-deficient mice.

Single nucleotide polymorphisms (SNPs) in TMEM106B encoding the lysosomal type II transmembrane protein 106B increase the risk for frontotemporal lobar degeneration (FTLD) of GRN (progranulin gene) mutation carriers. Currently, it is unclear if progranulin (PGRN) and TMEM106B are synergistically linked and if a gain or a loss of function of TMEM106B is responsible for the increased disease risk of patients with GRN haploinsufficiency. We therefore compare behavioral abnormalities, gene expression patterns, lysosomal activity, and TDP‐43 pathology in single and double knockout animals. Grn −/−/Tmem106b −/− mice show a strongly reduced life span and massive motor deficits. Gene expression analysis reveals an upregulation of molecular signature characteristic for disease‐associated microglia and autophagy. Dysregulation of maturation of lysosomal proteins as well as an accumulation of ubiquitinated proteins and widespread p62 deposition suggest that proteostasis is impaired. Moreover, while single Grn −/− knockouts only occasionally show TDP‐43 pathology, the double knockout mice exhibit deposition of phosphorylated TDP‐43. Thus, a loss of function of TMEM106B may enhance the risk for GRN‐associated FTLD by reduced protein turnover in the lysosomal/autophagic system.

Functional recreation of age-related CD8 T cells in young mice identifies drivers of aging- and human-specific tissue pathology

Mitigating effects of aging on human health remains elusive because aging impacts multiple systems simultaneously, and because experimental animals exhibit critical aging differences relative to humans. Separation of aging into discrete processes may identify targetable drivers of pathology, particularly when applied to human-specific features. Gradual homeostatic expansion of CD8 T cells dominantly alters their function in aging humans but not in mice. Injecting T cells into athymic mice induces rapid homeostatic expansion, but its relevance to aging remains uncertain. We hypothesized that homeostatic expansion of T cells injected into T-deficient hosts models physiologically relevant CD8 T cell aging in young mice, and aimed to analyze age-related T cell phenotype and tissue pathology in such animals. Indeed, we found that such injection conferred uniform age-related phenotype, genotype, and function to mouse CD8 T cells, heightened age-associated tissue pathology in young athymic hosts, and humanized amyloidosis after brain injury in secondary wild-type recipients. This validates a model conferring a human-specific aging feature to mice that identifies targetable drivers of tissue pathology. Similar examination of independent aging features should promote systematic understanding of aging and identify additional targets to mitigate its effects on human health.

Intraosseous epidermoid cysts of adjacent digits in a dog

BackgroundIntraosseous epidermoid cyst (IEC) is a rare, non-neoplastic, pathology in animals and humans that most commonly affects the distal phalanx. In dogs, it is important to differentiate this lesion from malignant digital tumours causing bone lysis. In previous reports, IEC has been described to affect only a single digit at the time of diagnosis which is usually based on histopathology. This is the first case report to describe immunohistochemically confirmed IECs affecting simultaneously multiple digits.Case presentationA 4-and-a-half-year-old female spayed Great Dane was presented with a 2-month history of progressive swelling of the distal phalanx (PIII) of digits IV and V of the right pelvic limb. Eleven weeks prior to presentation, the dog had a low-grade cutaneous mast cell tumour completely excised from the craniolateral base of its left pinna. A history of trauma to 1 of the nails of the same pes 4 years prior to referral was also reported. Examination of the right pelvic limb identified firm non-painful swelling of PIII of digits IV and V, with concurrent deformation of the nails. Radiographs of the right pes obtained by the primary veterinarian identified an expansile lesion of PIII of digits IV and V. Computed tomography identified large expansile lesions of PIII of digits IV and V, with associated cortical thinning and soft tissue swelling. Neoplasia was considered the most likely radiographic diagnosis. Histopathology of Jamshidi bone biopsies was consistent with intraosseous epidermoid cyst, which was confirmed with immunohistochemistry. Amputation of PIII of digits IV and V at the level of mid-PII was performed as definitive treatment. No recurrence of the lesion occurred during the 10-month follow-up period.ConclusionsIntraosseous epidermoid cysts should be included in the differential diagnosis for expansile lesions affecting the canine digit. It is important to differentiate them from other digital lesions, with bone involvement, such as malignant digital tumours, which often require more extensive surgery for definitive treatment. The case herein highlights that this lesion can affect simultaneously multiple digits. Definitive diagnosis can be achieved by identification of keratin-producing epithelial cells on histopathology and confirmed by pancytokeratin labelling.

Zika virus-induced neuro-ocular pathology in immunocompetent mice correlates with anti-ganglioside autoantibodies

ABSTRACT A severe consequence of adult Zika virus (ZIKV) infection is Guillain-Barré Syndrome (GBS), where autoreactive antibodies attack peripheral and central nervous systems (CNS) resulting in neuro-ocular pathology and fatal complications. During virally induced GBS, autoimmune brain demyelination and macular degeneration correlate with low virus neutralization and elevated antibody-mediated infection among Fcγ-R bearing cells. The use of interferon-deficient mice for ZIKV studies limits elucidation of antibody-dependent enhancement (ADE) and long-term pathology (≥120 days), due to high lethality post-infection. Here we used immunocompetent BALB/c mice, which generate robust humoral immune responses, to investigate long-term impacts of ZIKV infection. A high infectious dose (1x106 FFU per mouse) of ZIKV was administered intravenously. Control animals received a single dose of anti-IFNAR blocking monoclonal antibody and succumbed to lethal neurological pathology within 13 days. Immunocompetent mice exhibited motor impairment such as arthralgia, as well as ocular inflammation resulting in retinal vascular damage, and corneal edema. This pathology persisted 100 days after infection with evidence of chronic inflammation in immune-privileged tissues, demyelination in the hippocampus and motor cortex regions of the brain, and retinal/corneal hyperplasia. Anti-inflammatory transcriptional responses were tissue-specific, likely contributing to differential pathology in these organs. Pathology in immunocompetent animals coincided with weakly neutralizing antibodies and increased ADE among ZIKV strains (PRVABC59, FLR, and MR766) and all Dengue virus (DENV) serotypes. These antibodies were autoreactive to GBS-associated gangliosides. This study highlights the importance of longevity studies in ZIKV infection and confirms the role of anti-ganglioside antibodies in ZIKV-induced neuro-ocular disease.

Thoughts on Convergence Science of high-risk animals responsible for zoonotic epidemics

Since the beginning of the 21st Century, large viral outbreaks have threatened human health, economies, and biosecurity. On April 20, 2020, World Health Organization(WHO) reported more than 2.2 million confirmed cases and 150000 deaths from a novel coronavirus (Coronavirus disease 2019, COVID-19; strain SARS-CoV-2). According to a macroeconomic forecast from Standard & Poor’s Global Ratings, the global GDP will fall 2.4% this year, and the economic impact of COVID-19 is sure to be far-reaching. It is suspected that COVID-19 has a wildlife origin. Indeed, most emerging human infectious diseases, such as SARS, Ebola, and H1N1, originate in animals. However, animals that host infectious diseases not only play critical roles in disease transmission, control, and prevention, but also serve the basis for the maintenance and stability of natural ecosystems. Although the COVID-19 and other epidemic diseases remind us that information on animals and their pathogenic microbes is necessary to control the spread of zoonotic diseases before they turn into epidemics or pandemics, we have little knowledge on the behavior, ecology, life history, and the pathology of infected animals. Therefore, comprehensive and solid research should be performed on animals that carry pathogens (e.g., viruses, bacteria, parasites, fungi, and prions). We searched published articles on infectious, endemic, and epidemic diseases in the Web of Science database and found that only 11.75% of articles (2930727 in total) were in the field of zoology. In contrast, more than half of the articles were in infectious diseases (biomedicine). Chinese researchers have published 221105 papers, 26.72% of the output of the United States. Even in the field of zoology, the US has published 86233 articles (2.83 times more than Chinese researchers). As solutions to the challenges of zoonotic epidemics require a public and comprehensive approach, we propose to develop a Convergence Science approach (transdisciplinary science) to study high-risk animals responsible for zoonotic epidemics. It aims to answer and resolve basic but essential questions. For example, what is the biological background of these animals, and how many ways can zoonotic diseases spillover? This effort is not limited to species classification, biological characters, evolutionary footprints, hosted microbes, behavior, and spillover approaches, nor field investigation, genetics, molecular biology, physiological, microbiology, mathematics, management, and veterinary medicine/science. While we appreciate the efforts of the medical research community to develop vaccines and medications to mitigate the spread of emerging diseases and to reduce human mortality rates, we argue that generating a dynamic list of pathogenic microbes and their wildlife hosts also represents an urgent and critical tool in ecological epidemiology. We call for a global and cooperative effort to create, maintain, and update this information in order to reduce the severity of future pandemics. A Convergence Science approach to the study of animals that host potential infectious diseases may allow the prediction and prevention of zoonotic epidemics, reduce or ablate the risk of zoonotic infection, and ensure biosecurity and public health.

Physiological and biochemical studies as a necessary component of the algorithm for assessing the effectiveness of food minor biologically active substances

Preclinical physiological and biochemical studies of the effectiveness of the tested biologically active substances in the conditions of simulated pathology in laboratory animals are an important stage preceding the assessment of the clinical effectiveness of the created specialized foods for therapeutic or preventive nutrition. The aim of this work is a brief review of the developed algorithm for preclinical assessment of the effectiveness of specialized foods, which includes a set of sequential stages of testing the safety, stability, bioavailability and effectiveness of biologically active substances in experiments on laboratory animals. Results. The paper presents a brief review of methodological approaches for in vivo preclinical assessment of the effectiveness of minor plant biologically active substances - polyphenols from bilberry fruits and leaves and phytoecdysteroids extracted from spinach. A beneficial effect of bilberry leaves' polyphenols on carbohydrate and lipid metabolism of male Zucker and Wistar rats was shown, which determined the necessity to increase the effectiveness of the developed polyphenolic ingredient by sorption on a food carrier. When evaluating the in vivo effects of polyphenols from blueberries and leaves sorbed on buckwheat flour, a pronounced hypoglycemic effect was found. The experiments aimed to the assessment of the adaptogenic effects of phytoecdysteroids from spinach leaves showed a significant decrease in the content of the main biomarkers of the stress system, which indicates a «smoothing» of the response of the animal organism to a strong stress impact. Conclusion. The inclusion of minor biologically active substances into the composition of foods for preventive and therapeutic nutrition is the necessary condition to improve their effectiveness. The scientific and practical significance of the preclinical evaluation to a decisive extent depends on the presence of a correctly selected biomodel. Thus, the adequacy of the assessment of the effects of minor biologically active substances as functional food ingredients in the composition of the specialized food products depends on the reproducibility of pathological processes (clinical, biochemical and morphological disorders) characteristic for the studied disease on the selected in vivo model.

Effect of aging on immunogenicity and efficacy of inactivated influenza vaccines in cotton rats Sigmodon hispidus

ABSTRACT Inactivated influenza vaccines are known to be less immunogenic in human elderly in regards to serologic antibody response induced by vaccination. Accumulating evidence, however, points to a comparable effectiveness of influenza vaccines in the young and the elderly individuals. In the current study, we assessed immunogenicity and effectiveness of trivalent inactivated vaccine FluLaval in young and aged cotton rats Sigmodon hispidus and found that while serologic response to immunization was indeed reduced in older animals, comparable protection against influenza infection was afforded by prime-boost vaccination in both young and aged cotton rats. Both hemagglutination inhibition (HAI) titers and seroconversion rates were lower in the aged animals compared to the young ones. Reduction of viral load in the lung and nose, however, was comparable between young and aged animals vaccinated twice. One-time immunization with FluLaval was less efficacious at protecting the nose of aged animals, indicating that boosting of preexisting immunity can be particularly important for nasal protection in the elderly. Coincidentally, a one-time immunization with FluLaval had a detrimental effect on pulmonary pathology in the young animals, suggesting that boosting of immunity is essential for the young as well. Overall, these results suggest that reduced antibody response to and sufficient efficacy of influenza vaccines in the elderly are not two irreconcilable phenomena and that incomplete immunity to influenza can be detrimental at any age.

Obituary for Professor Michael J. Day

On 10th May 2020, Michael Day passed away at the age of 59 after a terminal illness with cancer. There have been many messages and tributes to him. He had many interests and activities as a veterinarian and was involved with the British Veterinary Small Animal Association (BSAVA), the World Small Animal Veterinary Association (WSAVA) as well as being the senior editor of the Journal of Comparative Pathology. There is an online book of condolence with many tributes at http://www.tolbc.com/MichaelDay. Michael trained as a veterinarian at Murdoch University, Perth, Western Australia. He had a profound interest in small animal immunology, infectious diseases and pathology which was focused in his residency and PhD studies performed at Murdoch. Having completed his PhD he held postdoctoral positions in Bristol and Oxford before joining the University of Bristol in 1990 as a lecturer in veterinary pathology in the Veterinary School. In 2017 he left Bristol Veterinary School as emeritus professor in veterinary pathology. Michael had been taught at Murdoch by Susan Shaw, who also joined the University of Bristol in 1990, and they forged a long-term collaborative friendship. For example, they co-authored a chapter entitled “Recent developments in atopic dermatitis of companion animals” (In: Williams HC, ed., Atopic dermatitis: the epidemiology, causes and prevention of atopic eczema. Cambridge: Cambridge University Press, 2000; 233–244). Michael was renowned for his ability to teach immunology to veterinary undergraduates and postgraduates by making the complex seem accessible. This ability was put to use helping residents of the European College of Veterinary Dermatology in their Immunology workshops in Split and ESVD members at a workshop in York. He lectured all over the world at international conferences about small animals and their immune-mediated and infectious diseases. He co-authored, with Ronald Schulz, a successful and popular textbook that has been widely used for teaching veterinary students (Veterinary immunology: principles and practice, 2nd edition. Boca Raton, FL: CRC Press, 2013). He also wrote two other books published by CRC Press, Clinical immunology of the dog and cat and (co-authored with Susan Shaw) Arthropod-borne infectious diseases of the dog and cat. Michael was extraordinarily hard-working and meticulously organised. For example, when compiling the book Veterinary Allergy (Chichester, UK: Wiley, 2014), the editors asked him to write the introductory chapter on “The immunological basis of allergic diseases”. His was, as expected, the first manuscript submitted, on time and complete with no revisions required. Michael worked on various techniques during his residency and PhD studies on the immune system of companion animals, and this is reflected in one of his earliest publications entitled “Immunodiagnosis of autoimmune skin disease in the dog, cat and horse” (Day MJ, Penhale WJ, Aust Vet J 1986; 63: 65–68). His research training in Murdoch led to the establishment of a diagnostic laboratory at the University of Bristol that enabled clinicians to investigate various diseases and conditions; it included testing for immunoglobulins, ANA titres and interpretation of immunohistochemical results for various tissues. This helped to bring him into contact with many clinicians within and outside Bristol Veterinary School and with whom he collaborated, this work helped to contribute to Michael’s large list of over 340 publications. While at Bristol Veterinary School, Michael helped to develop the application of reagents in detecting canine immunoglobulin G subclasses, and this led to a series of studies of immune responses in various skin diseases in dogs including pyoderma, hypersensitivity disorders and immune-mediated diseases. He also published detailed substantial retrospective histopathological studies looking at demodicosis, auto-immune skin diseases, pyoderma and cutaneous lymphoma. Michael will be remembered by many for his engaging and collaborative approach to all things concerning the study of infectious and immune-mediated diseases in companion animals, as well as a substantial portfolio of work on experimental autoimmunity. He supervised numerous PhD students and postdoctoral fellows, and was an external examiner for many. In later years Michael explored his family history and, interestingly, it became apparent that his family had hailed from the southwestern England, from villages in the county of Somerset, before they moved out to Australia in the 19th Century. Consequently, when he worked for the University of Bristol and made his home in Cheddar in the Mendip Hills area of Somerset, he was not far from where his ancestors had set out for Australia. Our thoughts and sympathies are extended to his wife Mary and to his family.

Combined Effect of Nicotine and Caffeine on Orthodontic Tooth Movement in Rats

Background: The individual effects of nicotine and caffeine have been reported in previous studies but their combined effect on tooth movement needs to be elucidated. The objective of this study was to evaluate the combined effect of nicotine and caffeine on the magnitude of orthodontic tooth movement (OTM) in rats.
 Material and Methods: This experimental study was conducted on Sprague-Dawley rats (Animal House and Pathology Laboratory; Post Graduate Medical Institute, Lahore) in the department of Orthodontics, de’Montmorency College of Dentistry, Lahore from 8th July 2014 to 8th January 2015. Forty male Sprague-Dawley rats were divided into four equal groups: Control group (CR), nicotine group (NT), caffeine group (CF) and combined nicotine and caffeine group (CNC). Closed coil nickel titanium (NiTi) spring was placed between incisor and maxillary molar. Nicotine group (NT) was treated by intraperitoneal injections of nicotine. Caffeine was given to caffeine group and Combined nicotine and caffeine group (CNC) was treated in the same way as individual nicotine and caffeine groups daily for 14 days. All the rats were sacrificed on 15th day. Magnitude of the orthodontic tooth movement was measured using digital Vernier caliper. Means and standard deviation were calculated for orthodontic tooth movement. One-way ANOVA was used to determine the mean difference in OTM. Post hoc Tukey test was used for multiple comparisons among the groups.
 Results: The mean orthodontic tooth movement (OTM) was 0.32 mm ± 0.05 in control group, 0.56 mm ± 0.04 in nicotine group, 0.52 mm ± 0.034 in caffeine group and 0.8 mm ± 0.06 in combined NC group, respectively. The difference between mean OTM among the groups was statistically significant (P-value <0.001). The mean OTM in CNC group was significantly higher as compared to other groups (CR, NT, CF, NT) (P-value <0.001).
 Conclusions: In rats, the combined use of nicotine and caffeine results in greater orthodontic tooth movement as compared to their individual use.

Variation in Gut Microbiota of Captive Bengal Slow Lorises.

Gastrointestinal microbiome plays an important role in animal metabolism, immune system and pathology associated with health and disease. Many wild slow lorises were confiscated from illegal trade into captivities and experienced a range of changes in living environment and diet. Microbiome analysis contributes to improving captive management by identifying the alteration in their gastrointestinal microbial communities and aiding in determining the factors affecting the health of captive slow lorises. The fecal samples of eighteen Bengal slow lorises (Nycticebus bengalensis) were used to compare gut microbiota from four rescue centers located in Dehong, Gejiu, Nanning and Puer cities of China. The results showed a significant site-dependent difference in microbial community diversity. Similar to other Lorisinae species, the Phyla including Bacteroidetes, Firmicutes and Proteobacteria dominated their gut microbiome composition. The Gejiu group exhibited a higher overall diversity and the unique OTUs, which is resulted from long-term isolated husbandry and heavy human disturbances. The scarcity of gums in the captive diet was likely to cause a lower abundance of Prevotella associated with soluble fiber degradation. The variation of intestinal microbiota in different environments highlights the necessity to improve feed preparation and husbandry management for the captive Bengal slow lorises.

Oral Fucoidan Attenuates Lung Pathology and Clinical Signs in a Severe Influenza a Mouse Model.

Fucoidans are known to be effective inhibitors of inflammation, and of virus binding and cellular entry. Undaria pinnatifida-derived fucoidan (UPF) was assessed in a severe influenza A (H1N1, PR8) infection model in mice. Initially, UPF was gavaged at 3.52 mg daily in a treatment model. Gross lung pathology (consolidation) was significantly reduced as compared to controls. UPF was then presented as a feed supplement at a rate of either nil, 3.52 mg/day or 7.04 mg/day in a prophylactic model, dosed three days before infection. A significant improvement was observed in the clinical signs of ill-health, as well as a reduction in gross lung pathology in animals treated with the higher dose, although there was no significant reduction in lung viral titres.

Vitamin D effectiveness and pathology in humans and domestic animals

Vitamin D is a fat-soluble vitamin that involved mainly for intestinal absorption of calcium with the regulation of calcium/phosphorus hemostasis, bone formation and remodeling, and anti-inflammatory with immune-modulating advantages. Insufficient solar UVB irradiation and/or lack of vitamin D in diet could induce a deficiency of vitamin D in animals and humans that manifested by poor appetite and growth and consequently leads to osteomalacia and osteodystrophy. A wide range of non-skeletal disorders, neoplastic, cardiovascular, and auto-immune diseases are attributed to vitamin D deficiency. This current review is presenting an overview of the origin/source, metabolism and activation, biology, physiology and functions, and the pathology of vitamin D in human and domestic animals. Special topics of vitamin D deficiency, toxicity, and rickets are also detailed in this review. In conclusion, maintenance of a normal serum level of active form of vitamin D prevents the occurrence of broad ranges of disorders and diseases. Beside enhance the weight gain and the growth of the growing animals, normal levels of vitamin D may also play a crucial role as immune-modulating factors for making animals and people resistant against contagious diseases such as hemorrhagic septicemia and coronavirus (COVID-19).