Diagnosis Of Animal Diseases Research Articles

Development and application of a novel Bio–Plex suspension array system for high–throughput multiplexed nucleic acid detection of seven respiratory and reproductive pathogens in swine

The aim of this study was to develop a multiple PCR assay based on the suspension array system for the simultaneous detection of respiratory and reproductive pathogens in swine. Pseudorabies virus (PRV), Japanese encephalitis virus (JEV), classic swine fever virus (CFSV), African swine fever virus (ASFV), porcine circovirus type 2 (PCV–2), porcine reproductive and respiratory syndrome virus (PRRSV) and porcine parvovirus (PPV) are the major respiratory and reproductive viral pathogens in pig farms. Seven pairs of specific primers and probes were designed, and the multiple PCR was performed, with the PCR products hybridized to beads coupled to probes, which were then detected by Bio–Plex suspension array system. The limit of detection, specificity and repeatability of this method was determined. The assay was further tested using 137 clinical samples, and the results were compared with conventional PCR to evaluate the ability of the method to diagnose porcine viruses. The results showed that the assay had a high degree of specificity and repeatability, and the simultaneous detection limit for the seven viruses reached 103 copies/μL. Forty-nine of the clinical samples tested positive for at least one of the viruses, the principal viral infections in the clinical samples were PCV–2 and PRRSV. The suspension method represented a rapid, specific and high–throughput tool for single or mixed detection of the seven porcine viruses simultaneously, and has great significance for the development of liquid chip techniques for the diagnosis of diseases in animals.

Biotechnological research in Ukraine for solving energy, medical-biological and medical-ecological problems in 2009-2015 taking into account the implementation of the association agreement with the EU

This article summarizes the main results of the biotechnological research of the National Academy of Sciences of Ukraine aimed at solving energy, medical-biological and medical-ecological problems during the period from 2009 to 2015. The prospective bioresources and directions of implementation of the latest technologies of bioenergy conversion for the production of liquid biofuels and expansion of their use for biodiesel were determined. The latest medical and biological and bioengineering technologies for human health and the national economy, biologically active substances for human health, ecological and economic mechanisms of rational use, protection and monitoring of natural resources, new technologies for the efficient use of energy resources were presented in the article. The authors systematized the results of the current research of the National Academy of Science of Ukraine aimed at the development of modern methods for the prevention and diagnosis of diseases in humans and animals, modern methods of cell biotechnology and metabolic engineering for the creation of superproducts of biologically active substances, new forms of plants and microorganisms, the genetic design of improved microorganisms and lines of plant and animal cells for the development of medical and agricultural biotechnology. The authors proved that these directions of research fully correspond to the world-wide trends in research in biomedicine.

Three- dimensional reconstruction of the thorax in the mongoose (Herpestes ichneumon)

Three- dimensional (3D) reconstruction obtained by using multidetector computed tomography (MDCT) images have widely been used in anatomical studies. Thorax is one of the most important body cavities necessary for the protection of lungs and heart in mammals. Two adult mongooses (1 male, 1 female) obtained from traffic accidents were used in this study. The images obtained from MDCT were stacked and 3D reconstruction of thorax was performed by overlaying images using a 3D modeling software (Mimics 13.1). Some measurements of thoracic cavity, lungs and sternum were taken from the reconstructive images of mongoose and indexes were calculated from these measurements. The morphometric parameters were recorded for both sexes. From the study, it could be concluded that the thoracic cavity, lungs and sternum imagings and findings revealed by 3D modeling techniques can be utilized for anatomical training of wild animals. This study is expected to help in the diagnosis and treatment of thorax diseases in wild animals.

Antibacterial Efficacy of Silver Nanoparticles on Endometritis Caused by Prevotella melaninogenica and Arcanobacterum pyogenes in Dairy Cattle.

Bovine postpartum diseases remain one of the most significant and highly prevalent illnesses with negative effects on the productivity, survival, and welfare of dairy cows. Antibiotics are generally considered beneficial in the treatment of endometritis; however, frequent usage of each antibiotic drug is reason for the emergence of multidrug resistance (MDR) of the pathogenic microorganisms, representing a major impediment for the successful diagnosis and management of infectious diseases in both humans and animals. We synthesized silver nanoparticles (AgNPs) with an average size of 10 nm using the novel biomolecule apigenin as a reducing and stabilizing agent, and evaluated the efficacy of the AgNPs on the MDR pathogenic bacteria Prevotella melaninogenica and Arcanobacterium pyogenes isolated from uterine secretion samples. AgNPs inhibited cell viability and biofilm formation in a dose- and time-dependent manner. Moreover, the metabolic toxicity of the AgNPs was assessed through various cellular assays. The major toxic effect of cell death was caused by an increase in oxidative stress, as evidenced by the increased generation of reactive oxygen species (ROS), malondialdehyde, protein carbonyl content, and nitric oxide. The formation of ROS is considered to be the primary mechanism of bacterial death. Therefore, the biomolecule-mediated synthesis of AgNPs shows potential as an alternative antimicrobial therapy for bovine metritis and endometritis.

Resolving Ethical Dilemmas in a Tertiary Care Veterinary Specialty Hospital: Adaptation of the Human Clinical Consultation Committee Model

Technological advances in veterinary medicine have produced considerable progress in the diagnosis and treatment of numerous diseases in animals. At the same time, veterinarians, veterinary technicians, and owners of animals face increasingly complex situations that raise questions about goals of care and correct or reasonable courses of action. These dilemmas are frequently controversial and can generate conflicts between clients and health care providers. In many ways they resemble the ethical challenges confronted by human medicine and that spawned the creation of clinical ethics committees as a mechanism to analyze, discuss, and resolve disagreements. The staff of the North Carolina State University Veterinary Hospital, a specialty academic teaching institution, wanted to investigate whether similar success could be achieved in the tertiary care veterinary setting. We discuss the background and rationale for this method, as well as the approach that was taken to create a clinical ethics committee.

Acute phase response in experimentally infected broilers with avian infectious bronchitis virus serotype 4/91

Measurement of acute phase proteins is important for diagnosis of animal diseases. In the present study, effects of avian infectious bronchitis virus infection on acute phase response and acute phase proteins were determined. Thirty one-day-old commercial broiler chicks were reared in experimental facilities and, at the age of 21 days, were challenged intranasally with 0.2 mL of allantoic fluid virus suspension (titre 106.5 EID50 per 0.1 mL). Serum samples were obtained prior to challenge and on days 1, 2, and 5 post-inoculation. Haptoglobin, serum amyloid A and C- reactive protein concentrations were measured. Our results showed that all investigated acute phase proteins increased significantly after infection, with mean maximum concentrations between 24 h and 48 h. No correlation was observed between plasma acute phase proteins in the chickens prior and post inoculation of the virus. Haptoglobin was most sensitive factor to change in the exposed birds.

Serum activities of two bone markers in captive Asian elephants (Elephas maximus) at different ages.

The blood biochemical analysis of bone markers could have a role in the early diagnosis of metabolic bone disease in animals; however, there is limited information on bone markers in captive Asian elephants (Elephas maximus). Serum samples from ten captive Asian elephants were obtained to clarify the relationship between age and the blood bone markers tartrate-resistant acid phosphatase isoform 5b (TRAP5b) and bone specific alkaline phosphatase (BALP). Serum TRAP5b and BALP activities were negatively correlated with age. A positive correlation was observed between TRAP5b activity and BALP activity. These results may contribute to the health management of captive Asian elephants.

Експрес-оцінка репродуктивної здатності псів в умовах розплідника

In recent decades doctors of veterinary medicine have devoted particular attention to dogs and, in particular, to the questions of physiology and pathology of their reproduction. The result is the publication of a significant number of manuals and monographs devoted to the actual issues of diagnosis and treatment of small animal diseases. Little remains the problems of andrological pathology of dogs, while the requirements for quality and number of offspring in high-value breeds of dogs and, consequently, reproductive health dogs. An important direction in solving these issues is the holding of andrological dispensary, which allows to diagnose the form of impotence, to identify the causes, to develop medical and preventive measures. However, in the conditions of large nurseries to study a significant number of dogs in limited time, there is a need for an accelerated, but at the same time, objective assessment of reproductive capacity of males. Proceeding from this, the aim of the work was to validate the method developed by us for express evaluation of the reproductive capacity of dogs in the conditions of a particular nursery.We have developed an express assessment of the reproductive capacity of dogs, which involves conducting a general clinical trial of males, a survey of the genitourinary system, determining the manifestation of sexual reflexes and the withdrawal of spermograms. The obtained results of research were distributed on a scale: «G» (good) – pathological changes were not found, all indicators of homeostasis and spermograms within the norm. The pedigree can be used without restrictions; «S» (satisfactory) – slight deviations from the norm, which have a minor effect on the body and partly reduce the reproductive potential of the dog. The herb is allowed to reproduce with a restriction on the careful control of its reproductive use; «U» (unsatisfactory) – a pathological state of the body, hypotension or impotence. Breeding is not allowed.According to the results of the research, it was determined that express assessment of the reproductive capacity of the dogs makes it possible to carry out a quick and fairly objective assessment of the reproductive potential of a large number of dogs for a limited period, to identify diseased animals, to develop and implement the necessary preventive or curative measures.It was established that from the total number of evaluated dogs of the «Zvezdnoye schastye» nursery, 12 dogs (or 76%) were rated «good» and recommended for breeding, 4 ‒ were «satisfactorily» (18%) and could be allowed to be restrained with careful control their reproductive use, 1 ‒ «unsatisfactory» (6%) with the recommendation to carry out the treatment of concomitant pathologies and pass a repeat test in a month.

Simultaneous analysis of uric acid, xanthine and hypoxanthine using voltammetric sensor based on nanocomposite of palygorskite and nitrogen doped graphene

Abnormal level of purines, especially uric acid (UA), xanthine (XA) and hypoxanthine (HX) in organisms is an important marker for early diagnosis of animal (human) disease. In this paper, an simultaneous analysis of UA, XA and HX in biological samples was successfully realized using the fabricated voltammetric sensor based on the nanocomposite of both nano-rod clay mineral palygorskite (Pal) and two-dimensional nano-layered nitrogen doped graphene (NG). A water-soluble carboxymethyl cellulose (CMC) salt was simultaneously decorated both Pal and NG to improve the water dispersibility, and their nanocomposite was characterized. Under optimized conditions, The as-prepared nanocomposite electrode (CMC-Pal-NG/GCE) could realize their simultaneous voltammetric analysis in linear ranges from 0.5 to 60μM for UA, 0.2 to 20μM for XA, and 0.6 to 55μM for HX. The limit of detection of three purines (UA,XA and HX) is calculated to be 0.1 for UA, 0.05 for XA and 0.4μM for HX, and their possible detailed mechanisms for electro-catalytic oxidation and chemical reaction was discussed, which was successfully empolyed for simultaneous voltammetric analysis of three purines in real samples with good reproducibility, repeatability and stability. Satisfactory results indicated that the obtained nanocomposite will provide a promising candidate for the simultaneous electrochemical determination of UA, XA and HX in analysis of practical samples.

Perfil hematológico del burro criollo (Equus asinus) colombiano

El diagnóstico de las enfermedades en los animales, depende en gran medida de los exámenes clínicos y resultados de laboratorio, en los cuales se reflejan variaciones biológicas. Los valores hematológicos son muy importantes para la evaluación del estado fisiológico normal de un animal. Sin embargo, son muy pocos los trabajos que se han llevado a cabo en este aspecto en asnos criollos. El objetivo de esta investigación fue determinar los valores medios de un hemograma para burros criollos del tópico bajo colombiano. Se realizó un estudio descriptivo de corte transversal con muestro no probabilístico por conveniencia. Se tomaron muestras de 85 burros machos entre 4 y 15 años clínicamente sanos y alimentados con pasturas de Brachiaria humidicula. Estos parámetros se obtuvieron por medio de un analizador hematológico (ABX Micros ESV 60). Los valores obtenidos fueron: leucocitos 7,6±1,6 x103/mm3, linfocitos 39,69±10,87%, monocitos 5,89±1,63%, granulocitos 54,41±10,75%, eosinófilos 7,08±2,77%, eritrocitos 4,48±0,85 x106/mm3, hemoglobina 8,62±1,3 g/dL, hematocrito 30,24±5,1%, volumen corpuscular medio 67,71±5,91 µm3, hemoglobina corpuscular media 19.45±1.94 pg, concentración de hemoglobina corpuscular media 28,48±1,36 g/dL y plaquetas 354,20±136,24 x103/mm3. Los resultados confirman que existen diferencias entre los valores en la mayoría de los parámetros hematológicos de burros criollos comparados con los de otras razas y equinos. Estos resultados pueden ser útiles en la rutina clínica veterinaria para el diagnóstico, seguimiento y control de enfermedades.

Plasma proteome of buffaloes.

The proteomic approach has aroused the interest of veterinary medicine researchers, especially regarding the production of biopharmaceuticals and diagnosis of diseases in farm animals. Water buffaloes have gained prominence in the world economy due to the quality of their milk, meat, and leather, in addition to being an important donor of blood components. This work aimed to identify and characterize the proteins present in the blood plasma of Murrah buffaloes (Bubalus bubalis) through 2D electrophoresis, in gel protein digestion followed by mass spectrometry technique and for albumin depletion, in solution protein digestion followed by shotgun analysis. Our results showed the identification of 112 protein spots and 35 individual proteins, respectively. The abundant proteins were represented by albumin, fibrinogen-α, fibrinogen-β, fibrinogen-γ, immunoglobulins in general, α-1-antiproteinase, α-1B-glycoprotein, α-2-HS-glycoprotein, α-macroglobulin, apolipoprotein A1, antithrombin-III, endopin 2B, fetuin-B, retinol-binding protein, serotransferrin, transthyretin and vitamin D-binding protein. Most of these proteins are related to the signaling pathways of the complement system and coagulation cascade. The results allowed a better understanding of the protein composition of these blood components, thus promoting studies on animal health in the search for molecular markers of zoonotic diseases in buffaloes.

Livestock metabolomics and the livestock metabolome: A systematic review.

Metabolomics uses advanced analytical chemistry techniques to comprehensively measure large numbers of small molecule metabolites in cells, tissues and biofluids. The ability to rapidly detect and quantify hundreds or even thousands of metabolites within a single sample is helping scientists paint a far more complete picture of system-wide metabolism and biology. Metabolomics is also allowing researchers to focus on measuring the end-products of complex, hard-to-decipher genetic, epigenetic and environmental interactions. As a result, metabolomics has become an increasingly popular “omics” approach to assist with the robust phenotypic characterization of humans, crop plants and model organisms. Indeed, metabolomics is now routinely used in biomedical, nutritional and crop research. It is also being increasingly used in livestock research and livestock monitoring. The purpose of this systematic review is to quantitatively and objectively summarize the current status of livestock metabolomics and to identify emerging trends, preferred technologies and important gaps in the field. In conducting this review we also critically assessed the applications of livestock metabolomics in key areas such as animal health assessment, disease diagnosis, bioproduct characterization and biomarker discovery for highly desirable economic traits (i.e., feed efficiency, growth potential and milk production). A secondary goal of this critical review was to compile data on the known composition of the livestock metabolome (for 5 of the most common livestock species namely cattle, sheep, goats, horses and pigs). These data have been made available through an open access, comprehensive livestock metabolome database (LMDB, available at http://www.lmdb.ca). The LMDB should enable livestock researchers and producers to conduct more targeted metabolomic studies and to identify where further metabolome coverage is needed.

Аналіз результтів лабораторних досліджень щодо бактеріозів у Харківській області

In the structure of gross agricultural output, livestock production is more than 38%. One of the main tasks of Veterinary Medicine is the implementation of the state veterinary and sanitary control and supervision over the observance of sanitary legislation, animal health and welfare, security and individual indicators of quality of food, non-food products of animal origin, reproductive material, etc. One of the most important links in the maintenance of epizootic well-being, animal health and indirectly people is the timely diagnosis of infectious animal diseases in order to prevent economic losses and disease outbreaks among consumers of livestock products. The article presents data on the results of laboratory studies of biological material from different species, and data on the pathogens of bacterial diseases in the territory of the Kharkiv region. The attention is focused on substantially reducing the amount of biological material coming for diagnostic studies, which correlates with the negative dynamics of the number of productive animals. Most of the isolated biological material Escherichia coli (colibacillosis exciter and swine edema disease) – 61.7% of cases, Salmonella spp. (The causative agent of salmonellosis and pulloroza birds) – 23.3% of cases, Streptococcus pneumoniae (pathogen diplokokoza / streptococcosis) – 5.1% of the cases, Pseudomonas aeruginosa (the causative agent psedomonoza) – 3.4% of cases. Furthermore, by bacteriological method was diagnosed: swine erysipelas, infectious epididymitis sheep, pasteurellosis, leptospirosis, campylobacteriosis manufacturers bulls stafilokokkoz, аmerican foulbrood bees, bees еuropean foulbrood listeriosis, braxy, malignant edema, tuberculosis poultry, fish and аeromonas, anthrax.

Molecular typing and diagnosis of Anaplasma spp. closely related to Anaplasma phagocytophilum in ruminants from Tunisia

Accurate diagnosis of animal and zoonotic diseases, such as granulocytic anaplasmosis, is crucial to estimate risk during control programs. In this study, 16S rRNA nested PCR and RFLP assay were combined to investigate the presence of Anaplasma phagocytophilum and genetically related strains (namely A. phagocytophilum-like 1 and 2) in 936 Tunisian ruminants. By using this method, A. phagocytophilum was not detected in any of the tested animals, while A. phagocytophilum-like 1 and A. phagocytophilum-like 2 were detected at variable prevalence rates in sheep, goats and cattle at coinfection rates respectively of 3.9, 2.5 and 0.5%. Sequence analysis validated RFLP data, and confirmed the co-occurrence of two potentially novel species closely related to A. phagocytophilum in Tunisian ruminants. Phylogeny indicated the presence of genetic variants shared by different ruminant species for each type of A. phagocytophilum-like strains. Results raise concern on the use and interpretation of indirect and direct tests traditionally employed for detecting pathogenic A. phagocytophilum strains in ruminants and in other vertebrates’ species, and provide additional background to improve classification of bacterial species closely related to A. phagocytophilum, and to reconstruct their evolutionary history.

A Novel Capillary Electrophoresis-Based High-Throughput Multiplex Polymerase Chain Reaction System for the Simultaneous Detection of Nine Pathogens in Swine.

Here we aimed to develop a capillary electrophoresis-based high-throughput multiplex polymerase chain reaction (PCR) system for the simultaneous detection of nine pathogens in swine. Nine pairs of specific primers and a set of universal primers were designed; the multiplex PCR was established. The specificity and cross-reactivity of this assay were examined, and the detection limit was determined using serial 10-fold dilutions of plasmids containing the target sequences. The assay was further tested using 144 clinical samples. We found that the nine specific amplification peaks were observed, and the assay had a high degree of specificity, without nonspecific amplification. The simultaneous detection limit for the nine viruses reached 10000 copies μL−1 when all of the premixed viral targets were present. Seventy-seven of the clinical samples tested positive for at least one of the viruses; the principal viral infections in the clinical samples were porcine circovirus type 2 and porcine reproductive and respiratory syndrome virus. This approach has much potential for further development of high-throughput detection tools for the diagnosis of diseases in animals.

Increasing importance of genomics in recognition of infectious animal diseases taking into account OIE requirements

Currently, the diagnosis methods applied in bacteriology and virology are frequently focused at modern molecular methods such as polymerase chain reaction (PCR), real-time PCR, loop-mediated isothermal amplification (LAMP) or confirmatory methods for already identified infectious agents of animals such as high throughput sequencing- HTS, or next-generation sequencing (NGS). A number of these methods are officially recommended by the World Organisation for Animal Health (OIE) for the routine diagnosis of major animal diseases with the greatest economical impact. The aim of this paper is to provide to a reader an overview on recent genomic methods used in diagnosis of infectious animal diseases along with their advantages and limitations. These methods facilitate efficient and reliable identification of animal pathogens and allow to characterize and sometimes to identify a newly recognized species of bacteria and viruses.

Ultrasonographic evaluation of the renal dimensions in captive tigers.

Ultrasonographic measurements of kidney size are useful in the practical diagnosis of kidney diseases in animals. In tigers, there is a lack of information regarding the ultrasonography methods used to measure the kidney size of the tiger. Thirty-three healthy captive tigers (Panthera tigris) were placed in lateral recumbency for ultrasonography. The measurements obtained from the ultrasonography were computed, and the results showed that there was a statistically significant difference between genders in terms of body weight and renal length. The length of the right kidney was significantly different from that of the left kidney (10.23 ± 0.76 cm in males versus 9.94 ± 0.80 cm in females; P<0.05). Interestingly, this study demonstrated that kidney length was statistically significantly associated with the body weight, and it also had a positive linear relationship with the body weight. Therefore, ultrasonographic renal dimensions could prove to be beneficial and modality for use in the evaluation of kidneys in unconscious tigers. However, kidney size evaluation must be performed using not only ultrasound but other clinical forms of technology and parameters.

Comparison of artificial neural network and K-means for clustering dairy cattle

Artificial neural network models (ANN's) are machine-learning systems, a type of artificial intelligence. They have been inspired by and developed along the working principles of the human brain and its nerve cells. ANN's are especially used in the modelling of nonlinear systems. With the information learned through repeated experience, similar to human learning, ANN's can provide classification, pattern recognition, optimisation and the realisation of forward-looking forecasts. Artificial neural network studies have been performed in animal husbandry in recent years. They have been used for the prediction of yield characteristics and classification, animal breeding, quality assessment, and disease diagnosis. In this study, classification of dairy cattle using artificial neural networks and cluster analysis are compared. Artificial neural networks models were determined to be more successful than cluster analysis.

Application of Microarray in Animal Disease Pathogenesis and Diagnosis

Microarray is a plethora of multiple genetic data. It is a collection of microscopic spots contains picomoles (10-12 moles) of a specific sequences (DNA, cDNA, cRNA, Oligonucleotides), known as probes. These probes hybridize with specific nucleic acid sequence called targets which are labelled with a fluorescent dye. Hybridization property between nucleotides were utilized in this and the signals produced are scanned and image data is converted to raw data using various softwares. Microarray has number of applications in the area of evolution biology, pathway analysis, toxicogenomics, pharmacogenomics, forensics, oncology and disease diagnostics and characterization. The genes responsible for several stages of cell to cell interaction of pathogens from adhesion to entry into the host cell and evading the host immune mechanism can be studied. Comparative genomic hybridization, not only yield information of individual genes in different tissues (normal or diseased, cancerous or non cancerous cells), but also on the regulation of different genes regulated pathways. Studying the molecular signature of pathogens paves an easy way for their detection. Moreover, these informations are utilized for better diagnostics, vaccine and drug development. Use of microarrays during outbreaks and/or disease surveillance would save time and help in early decisions to control the spread of disease. Thus, microarray proved to be a perfect drive in animal disease diagnostic. In veterinary field only few studies were done to interrogate the gene expression profile and detection of pathogens. While they have the potential, detection microarrays are yet to be used as routine surveillance tools in veterinary because of high sophistication, and cost per test is still high compared to other diagnostic methods. With all these advances many novel techniques born out of this technology with less cost and minimum training will play increasing role in disease diagnosis in near future.

PCR quantitativa (qPCR) no diagnóstico de doenças infecciosas em cães e gatos – revisão de literatura

Quantitative PCR (qPCR) has recently replaced conventional PCR and is becoming the molecular technique of choice, offered by the major laboratories of veterinary and human diagnostics worldwide. The aim of this review is to describe the usefulness of this technique for the diagnosis of infectious diseases in small animals. Through qPCR, one can investigate disease by either confirming the involvement of a specific agent or by testing the clinical sample for detection of a variety of agents by means of multiple-agent panels that are elaborated with the most likely pathogens for certain clinical signs of affected systems. Considering that a large number of infectious agents may be present in co-infections, understanding and detecting such agents may allow for adequate prognosis, treatment planning and prevention strategies in the clinical setting. Advantages and limitations of molecular tests are described in comparison to the routinely performed tests in the small animal clinics for diagnosis of infectious diseases in dogs and cats.