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PubMed2004

Staphylococcus aureus: study of genomic similarity of strains isolated in veterinary pathology using amplified fragment length polymorphism (AFLP).

Staphylococcus aureus is a widespread pathogen causing infections in different animal species. The extensive use of antibiotics, particularly methicillin, causes the rise of antibiotic-resistant strains (MRSA). In order to verify the epidemiology and genetic relatedness among MRSA and sensible strains (MSSA), an accurate fingerprinting technique, the amplified fragment length polymorphism (AFLP), was carried out. The isolates were cultured, subdivided on MRSA and MSSA and submitted for the genomic DNA extraction that was utilized for AFLP. The data were analysed for genetic similarity using the Dice coefficient. The results of genomic analysis among MRSA and MSSA and within them revealed that the major component of variation was due to variation within strains (82.12%), while variance among strains was lower (17.88%). The low level of genomic similarity found among S. aureus strains implies high level of genetic diversity. Different similarity was found as well in all strains independently of the source.

Animals↗
PubMed2005

Borna disease virus and the evidence for human pathogenicity: a systematic review.

BACKGROUND: Borna disease is a neurological viral disease of veterinary importance in central Europe, although Borna Disease virus (BDV) has been reported to be present in animals in most continents. The hypothesis that BDV is associated with human illness is controversial. However, should even a small fraction of mental illness be attributable to infection with BDV, this would be an important finding, not least because illness in that sub-population would, theoretically, be preventable. METHODS: We systematically reviewed the evidence: that BDV infects humans; for the role of BDV in human neuropsychiatric illness; to assess the suitability of currently available laboratory methods for human epidemiological studies. RESULTS: We identified 75 documents published before the end of January 2000, describing 50 human studies for BDV. There were five case studies and 44 (sero)prevalence studies, in a variety of patient groups. Nineteen prevalence studies (43%) investigated seroprevalence, 11 (25%) investigated viral prevalence and 14 (32%) investigated both. Seroprevalence ranged from 0% to 48%, and prevalence of virus or viral footprints from 0% to 82%. DISCUSSION: Although agreed gold standard tests and evidence for test specificity are lacking, there is evidence that humans are exposed to the virus. Further epidemiological studies are required to establish whether there are associations with disease.

Antibodies, Viral↗
PubMed2006

Survey of the large-animal diplomates of the American College of Veterinary Internal Medicine regarding knowledge and clinical use of polymerase chain reaction: implications for veterinary education.

A questionnaire was developed to document the knowledge base of large-animal diplomates of the American College of Veterinary Internal Medicine (ACVIM) regarding polymerase chain reaction (PCR) technology and to identify the common use of this technology in equine practice. Ninety-three of the 278 mailed questionnaires were returned, for an overall response rate of 33.4%. Ninety respondents (99%) reported being familiar with the general principles of nucleic acid probe technology; however, only 52 (57%) knew the difference between conventional (traditional) and real-time (second-generation) PCR. The majority of the respondents (88%) emphasized the need for continuing education on molecular diagnostics. Eighty-four (92%) of the respondents regularly use PCR (conventional and/or real-time) for the detection of equine pathogens, and 80 (88%) commonly submit their samples to university/state veterinary laboratories. Blood, nasal swabs, and feces are the three equine specimens most commonly submitted for PCR analysis of Streptococcus equi, Lawsonia intracellularis, Neorickettsia risticii, equine herpesvirus 1/4, Rhodococcus equi, Sarcocystis neurona, and equine influenza virus. Diplomates reported costs associated with molecular diagnostics and unreliability of PCR as the most common limitations of PCR. Didactic training in veterinary curricula and during continuing-education opportunities continues to be necessary to produce veterinarians who have an understanding of the clinical applications of molecular diagnostics.

Animals↗
PubMed2003

Recombinant Newcastle disease virus as a vaccine vector.

Veterinary vaccines remained conventional for more than fifty years. Recent advances in the recombinant genetic engineering techniques brought forward a leap in designing vaccines for veterinary use. A novel approach of delivering protective immunogens of many different pathogens in a single virus vector was made possible with the introduction of a "reverse genetics" system for nonsegmented negative-sense RNA viruses. Newcastle disease virus (NDV), a nonsegmented negative-sense virus, is one of the major viruses of economic importance in the poultry industry throughout the world. Despite the availability of live virus vaccines of good potency, the intrinsic ability of attenuated strains to revert in virulence makes control of this disease by vaccination difficult. Armed with the knowledge of virulence factors of this virus, it is now possible to produce genetically stable vaccines and to engineer mutations that enhance immunogenicity. The modular nature of the genome of this virus facilitates engineering additional genes from several different pathogens or tumor-specific antigens to design contemporary vaccines for animals and humans. This review will summarize the developments in using NDV as a vaccine vector and the potential of this approach in designing next generation vaccines for veterinary use.

Animals↗
PubMed1992

Immunostimulants added to injected Aeromonas salmonicida bacterin enhance the defense mechanisms and protection in rainbow trout (Oncorhynchus mykiss).

Immunostimulants were given to rainbow trout for assaying effects on modulating non-specific defense mechanisms, specific immune response, and protection levels against pathogen challenge with Aeromonas salmonicida. Three drugs, levamisole (an approved veterinary drug in the USA), a quaternary ammonium compound (QAC), and a short-chain polypeptide (ISK) were found to affect the non-specific defense mechanism activities, which were measured by changes in circulatory neutrophil and phagocytic activity levels, and the specific immune response factors, which were measured by numbers of plaque-forming cells, and circulatory antibody levels. When given alone, the immunostimulants elevated the non-specific factors. When injected in combination with an A. salmonicida O-antigen bacterin, the non-specific factors were further elevated, and the specific response was raised over samples taken from fish given the bacterin without the immunostimulants. Challenge tests with the virulent pathogen, A. salmonicida, showed a 5-6 day delay in the onset of mortalities in the fish given the immunostimulants alone, and a 12-14 day delay when immunostimulants given were combined with the bacterin. In the groups given the QAC or ISK with the bacterin, there was a 20% and 40% survival rate, respectively.

Adjuvants, Immunologic↗
PubMed2002

Impact of antimicrobial resistance on regulatory policies in veterinary medicine: status report.

Increasing resistance to antimicrobial agents is of growing concern to public health officials worldwide. The concern includes infections acquired in hospitals, community infections acquired in outpatient care settings, and resistant foodborne disease associated with drug use in food-producing animals. In the United States, a significant source of antimicrobial-resistant foodborne infections in humans is the acquisition of resistant bacteria originating from animals. The US Food and Drug Administration's (FDA's) goal in resolving the public health impact arising from the use of antimicrobial drugs in food-producing animals is to ensure that significant human antimicrobial therapies are not compromised or lost while providing for the safe use of antimicrobials in food animals. The FDA's approach to the problem is multipronged and innovative. The strategy includes revision of the pre-approval safety assessment for new animal drug applications, use of risk assessment to determine the human health effect resulting from the use of antimicrobials in food animals, robust monitoring for changes in susceptibilities among foodborne pathogens to drugs that are important both in human and veterinary medicine, research, and risk management.

Animals↗
PubMed2026

Genomic and phenotypic characterization of mcr-9 carrying Enterobacter oligotrophicus recovered from bovine mastitis.

Bovine mastitis (BM) is a leading cause of economic loss in the dairy industry, driven by decreased milk yields, involuntary culling, and substantial veterinary costs. A single Gram-negative isolate recovered from BM was characterized in this study. For this, antimicrobial susceptibility testing (AST) was performed using the Neg-Urine-Combo 98 panel. Whole-genome sequencing (WGS) was employed to identify antimicrobial resistance genes (ARG), virulence factors (VF) genes, plasmid replicons and prophage sequences. Comparative genomic analysis was performed through phylogenetic analysis. The BM isolate was phenotypically identified as Citrobacter rodentium, however, WGS analysis reclassified the isolate as Enterobacter oligotrophicus. Phenotypic AST revealed a resistance profile of 12%, with the isolate exhibiting resistance to β-lactams antimicrobials, specifically amoxicillin-clavulanate, ampicillin and the cephalosporin-cefoxitin. Conversely, susceptibility was demonstrated for the remaining tested antimicrobials. Genomic profiling identified 31 ARG, 10 VF genes and 6 prophage sequences within the E. oligotrophicus genome. This study provides the first evidence of E. oligotrophicus as a causative agent of BM, expanding the known spectrum of pathogens affecting the dairy industry by delivering the second complete genome of E. oligotrophicus available globally. The identification of 31 ARG, 10 VF, and 6 prophage sequences underscore the potential pathogenic risk and environmental resilience of this isolate. These findings highlight the critical role of WGS-based surveillance in identifying non-conventional mastitis pathogens and underscore the need for targeted mitigation strategies in veterinary medicine.

Animals↗
PubMed1975

Pathogenic mechanisms not operating in Eimeria necatrix infections.

Having investigated certain aspects of Eimeria necatrix coccidiosis in chickens, workers of the Hannover Veterinary School postulated "that death following a single inoculation of a large number of oocysts is due to an alarm reaction and not a specific pathogenic action of the parasites". Because this hypothesis is somewhat revolutionary in its consept, several pieces of evidence on which it is based and several logical deductions which can be made from it have been examined. It has been confirmed that injection of chichens with cysteamine or 5-hydroxytryptamine 30 min before inoculation of the birds with a lethal dose of E. necatrix oocysts reduces subsequent mortality; the reason for this, however, appears not to be the neutralisation of the proposed shock reaction, but rather an inhibition of the excystation process, brought about indirectly through the host. Inoculation of chickens with a non-lethal dose of E. necatrix oocysts 30 min before inoculation with a lethal dose of oocysts was followed by increased mortality rather than the decreased mortality which the hypothesis would predict. Treatment of chickens with sulphadimidine starting 48 h after inoculation resulted in survival of the birds rather than death which would ensue if in fact mortality was due to a shock reaction irreversibly initiated at the time of inoculation. A direct effect of sulphadimidine on the parasite has been shown both in vivo and in vitro.

Adrenocorticotropic Hormone↗
PubMed2002

Measuring tubulin content in Toxoplasma gondii: a comparison of laser-scanning confocal and wide-field fluorescence microscopy.

Toxoplasma gondii is an intracellular parasite that proliferates within most nucleated cells, an important human pathogen, and a model for the study of human and veterinary parasitic infections. We used a stable yellow fluorescent protein-alpha-tubulin transgenic line to determine the structure of the microtubule cytoskeleton in T. gondii. Imaging of living yellow fluorescent protein-alpha-tubulin parasites by laser-scanning confocal microscopy (LSCM) failed to resolve the 22 subpellicular microtubules characteristic of the parasite cytoskeleton. To understand this result, we analyzed sources of noise in the LSCM and identified illumination fluctuations on time scales from microseconds to hours that introduce significant amounts of noise. We confirmed that weakly fluorescent structures could not be imaged in LSCM by using fluorescent bead standards. By contrast, wide-field microscopy (WFM) did visualize weak fluorescent standards and the individual microtubules of the parasite cytoskeleton. We therefore measured the fluorescence per unit length of microtubule by using WFM and used this information to estimate the tubulin content of the conoid (a structure important for T. gondii infection) and in the mitotic spindle pole. The conoid contains sufficient tubulin for approximately 10 microtubule segments of 0.5-microm length, indicating that tubulin forms the structural core of the organelle. We also show that the T. gondii mitotic spindle contains approximately 1 microtubule per chromosome. This analysis expands the understanding of structures used for invasion and intracellular proliferation by an important human pathogen and shows the advantage of WFM combined with image deconvolution over LSCM for quantitative studies of weakly fluorescent structures in moderately thin living cells.

Algorithms↗
PubMed2003

Molecular genetic methods in the veterinary clinical bacteriology laboratory: current usage and future applications.

In the last 5 years, numerous molecular methods have been published for the detection and characterization of bacteria in the field of veterinary medicine. PCR has been the most commonly used technology. Although not currently used for clinical veterinary diagnosis, new technologies such as liquid-phase hybridization, real-time PCR, pathogen load determination and DNA/protein microarray have been described and have many possible applications in the clinical bacteriology laboratory because of their sensitivity and efficiency. This review describes the basic principles and application of recently published DNA-based molecular techniques for the purpose of veterinary clinical bacteriological diagnosis. It covers advances in probe hybridization technology, DNA/RNA amplification techniques and other molecular detection methods, including 16S rRNA analysis for bacterial characterization and DNA microarrays for bacterial detection. The review briefly summarizes the application of molecular methods for the diagnosis of specific important bacterial infections of animals, and for other animal pathogens that are slow or difficult to isolate in the clinical bacteriology laboratory. In addition, the molecular detection of antimicrobial resistance genes and of bovine mastitis pathogens is briefly described and current commercially available tests are listed.

Animals↗
PubMed1999

[Infections and bacterial resistance in the community].

The frequency of resistance to antibiotics among community acquired pathogens and the number of drugs to which they are resistant are increasing world wide. Antimicrobial resistance in the Netherlands is still low. Resistance to antimicrobial drugs is clearly linked to consumption of antibiotics within and outside of the hospital. Use of antibiotics in veterinary medicine may also contribute to the occurrence of antimicrobial resistance in human pathogens. Strategies to limit the spread of resistant strains should include encouraging the judicious use of antimicrobial agents. Guidelines for antibiotic therapy should be based on results derived from well designed surveillance studies.

Animals↗
PubMed2004

The enteric toxins of Clostridium perfringens.

The Gram-positive pathogen Clostridium perfringens is a major cause of human and veterinary enteric disease largely because this bacterium can produce several toxins when present inside the gastrointestinal tract. The enteric toxins of C. perfringens share two common features: (1) they are all single polypeptides of modest (approximately 25-35 kDa) size, although lacking in sequence homology, and (2) they generally act by forming pores or channels in plasma membranes of host cells. These enteric toxins include C. perfringens enterotoxin (CPE), which is responsible for the symptoms of a common human food poisoning and acts by forming pores after interacting with intestinal tight junction proteins. Two other C. perfringens enteric toxins, epsilon-toxin (a bioterrorism select agent) and beta-toxin, cause veterinary enterotoxemias when absorbed from the intestines; beta- and epsilon-toxins then apparently act by forming oligomeric pores in intestinal or extra-intestinal target tissues. The action of a newly discovered C. perfringens enteric toxin, beta2 toxin, has not yet been defined but precedent suggests it might also be a pore-former. Experience with other clostridial toxins certainly warrants continued research on these C. perfringens enteric toxins to develop their potential as therapeutic agents and tools for cellular biology.

Amino Acid Sequence↗
PubMed1988

Loa loa - a neglected filaria.

There has been considerable research on the biochemistry and immunology of many human, veterinary and laboratory animal filariae, with a view to understanding pathogenic and protective immune mechanisms and developing ways to boost them(1-3). But, apart from a few isolated reports (eg. Refs 4,5), these aspects have been largely ignored for the human filarial parasite Loa loa. Obviously not all filariae can be studied in equal depth - almost 100 different life cycle patterns are documented(6) - but, as Margaret Pinder shows here, L. loa is an important human pathogen of considerable biological interest.

Journal Article↗
PubMed2001

Detection of Babesia caballi and Babesia equi in Dermacentor nuttalli adult ticks.

Ticks play an important role in human and veterinary medicine particularly due to their ability to transmit protozoan pathogens. In this study we have demonstrated that polymerase chain reaction (PCR) and nested PCR methods enabled detection of Babesia caballi and Babesia equi in field isolates of Dermacentor nuttalli adult ticks from Mongolia. Primers specific for 218 bp fragment merozoite antigen 1 (EMA-1) gene of B. equi successfully amplified products from all samples of D. nuttalli adult ticks while primers for the 430 bp fragment product from BC48 gene of B. caballi amplified products from seven of the 54 samples. Using PCR and nested PCR methods we have found mixed infections with B. equi and B. caballi in the tick vector. The amplified DNA fragment from D. nuttalli ticks was inserted into the EcoRV site of pBluescript SK and sequenced. The sequence of the 430 bp fragment was completely identical to the nucleotide sequence of the USDA strain of B. caballi. These results suggest that D. nuttalli may play an important role as a vector of both B. caballi and B. equi and also may be important in maintaining endemicity of equine piroplasmosis in Mongolia.

Animals↗
PubMed2003

Impact of travel on international spread of antimicrobial resistance.

Antimicrobial resistance, an escalating problem worldwide, affects a broad range of human diseases. Excessive and inappropriate drug usage is the key driver for the emergence of resistant organisms. Travel, trade and mass migration form an important mode for their spread. The use of molecular biology provides the means of understanding the genesis and spread of the genes for drug resistance. Antimicrobial use in veterinary practice as food additives causes selection of resistant zoonotic pathogens that may spread to humans. Comprehensive surveillance systems should be designed and implemented at local and national levels and a national resistance surveillance database operationalized. There is also need for better regulation of the use of antibiotics and education of the medical fraternity, veterinarians and the public in the appropriate use of antimicrobials.

Animals↗
PubMed2004

Emerging diseases and implications for global trade.

Emerging diseases could have a devastating impact on international trade unless there is a change in the traditional approach to disease control and new holistic prevention and control strategies are adopted. The impact that emerging diseases will have on international trade will depend on several factors, such as the nature of the pathogen, the degree of co-ordination and integration between Veterinary Services and Public Health authorities, the ability to rapidly detect and respond to a disease appearance, and the existing trade relationship between countries. Strategies to control emerging diseases will be more effective if competent authorities and veterinary infrastructures in particular, integrate their public and animal health objectives into a single strategy. The co-ordinating role of international standard setting organisations such as the World Organisation for Animal Health (OIE) will be crucial. This paper discusses the events contributing to the appearance and spread of new emerging diseases and examines the changing roles of Veterinary Services and international organisations. Reference is also made to measures that can be taken towards minimising potential trade disruptions caused by the appearance of an emerging disease.

Animals↗
PubMed2006

International organisations and their role in helping to protect the worldwide community against natural and intentional biological disasters.

Preventing the spread of disease through international movements is one of the key objectives of the World Organisation for Animal Health (OIE). One of the ways it seeks to achieve this is by publishing international standards and guidelines aimed at, inter alia, preventing the importation of pathogens that are dangerous for animals and humans and strengthening Veterinary Services so that they can improve their surveillance and response systems. The OIE works in close partnership with the Food and Agriculture Organization of the United Nations (FAO), and together the two organisations have developed a joint initiative - the Global Framework for the Progressive Control of Transboundary Animal Diseases (GF-TADs). Member Countries of these organisations could increase their capacity to manage the risks of disease occurrences, whether natural or deliberately introduced, if they would all strictly implement existing OIE international standards. Compliance with these standards greatly depends on the political willingness of national policy-makers and on a successful transfer of resources to developing countries in support of good governance and appropriate policy implementation. A United Nations Resolution obliging its Member Countries to implement OIE standards could prove invaluable in this respect.

Animal Diseases↗
PubMed2006

[The three types of influenza].

The unprecedented spread of the highly pathogenic avian influenza type A H5N1 is a threat to veterinary public health. However, the virus does not in its present form constitute a major threat to human public health. Only if the virus mutates to spread effectively from person to person will there be a risk of a new pandemic. It is not possible to predict if this will happen, when it may happen or how serious it may be. At present, seasonal influenza represents the greatest burden of illness among the three types of influenza A illness: seasonal, avian, and pandemic influenza.

Animals↗