Editorial: Sequencing technologies in advancing veterinary and zoonotic infection research.
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antimicrobial resistance↗
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Characterization of Listeria monocytogenes isolates from food animal clinical cases: PFGE pattern similarity to strains from human listeriosis cases.
Twenty-one isolates of Listeria monocytogenes from food animal clinical cases that involved meningitis or meningoencephalitis, encephalitis, mastitis and abortion were characterized by serotyping and pulsed-field gel electrophoresis (PFGE) in order to improve our understanding of the genetic links between individual strains and strains recovered from human listeriosis cases. Results showed that five of the isolates were serotype 1/2a, six were 1/2b, nine were 4b, and one was untypeable. A caprine, two bovine and an ovine brain isolate shared identical PFGE patterns indicating that strains of L. monocytogenes are not host specific. Other isolates exhibited distinct patterns that were not shared, indicating a genetic diversity. Dendrogram analysis revealed that PFGE patterns of the isolates clustered primarily according to serotype. We compared the PFGE types obtained for these isolates with PFGE types for human clinical isolates present in the CDC national PulseNet database. Six (29%) of the twenty-one strains had patterns that were indistinguishable from pathogenic human isolates in the database. Our observations offer preliminary evidence that food animals could be significant reservoirs of L. monocytogenes that lead to human infections and support the inclusion of PFGE patterns of veterinary clinical isolates in the national PulseNet database for increased surveillance.
Animals↗
Progress toward molecular characterization of ectoparasite modulation of host immunity.
Ectoparasitic arthropods and vector-borne infectious agents are global medical and veterinary public health concerns. Economic impact due to direct effects of infestation and disease transmission are significant. These problems are increased by development of arthropod resistance to insecticides/acaricides; drug resistance of vector-borne pathogens; and, lack of effective vaccines to prevent many of these diseases. There is much to be gained from understanding the complex array of immunological interactions occurring at the arthropod-host-pathogen interface. One application of that knowledge is the development of novel vaccines for the control of both ectoparasitic arthropods and the diseases they transmit. We now realize that blood-feeding arthropods are not simply flying or crawling hypodermic needles and syringes. Ectoparasitic arthropods are not passive partners in their relationships with the immune systems of their hosts. These clever invertebrates produce numerous pharmacologically active molecules that help them migrate through tissues of their hosts or to successfully obtain blood meals. Arthropod parasites stimulate a spectrum of host immune responses that could potentially impair development, reduce feeding success, or kill the ectoparasite. Not unexpectedly, arthropods have developed sophisticated arsenals of countermeasures that modulate or deviate host immune responses. Not only does arthropod modulation of host immunity facilitate survival in tissues or increase the likelihood of obtaining a blood meal, but it is increasingly recognized as a critical factor in pathogen transmission. Those countermeasures to host immune defenses are the topics of this review. Emphasis is placed on our current understanding of the molecular bases of those changes; the molecules responsible for host immunomodulation; contemporary approaches for studying these complex relationships; and, the potential for using this information to develop innovative vaccine-based control strategies.
Animals↗
[Anaplasmosis in a Swiss dairy farm: an epidemiological outbreak investigation].
In summer of 2002, a case of severe clinical bovine anaplasmosis caused great losses in a dairy farm of an animal trader in Grisons. This article outlines the general approach of an outbreak investigation considering the case of anaplasmosis as an example. The goals of such investigations are to identify and eliminate the source of a disease outbreak in order to avoid additional cases. In addition, recommendations should be developed for preventing or limiting the magnitude of similar outbreaks in the future. In the outbreak presented, the causative agents were probably brought into the dairy farm by animal trade. Due to the large herd size, a missing quarantine for new animals and the coinfection with several pathogens, this case led to a high number of fatalities. The investigations of this outbreak demonstrated the importance of an universal and consistent identification of individual animals for the reconstruction of their movements. The veterinary practitioner should be reminded to act cautiously when facing strange clinical cases and to also consider "exotic diseases" as a possible cause.
Anaplasmosis↗
West Nile virus vaccines.
West Nile virus (WNV) is a mosquito-borne flavivirus that is emerging as a global pathogen. In the last decade, virulent strains of the virus have been associated with significant outbreaks of human and animal disease in Europe, the Middle East and North America. Efforts to develop human and veterinary vaccines have taken both traditional and novel approaches. A formalin-inactivated whole virus vaccine has been approved for use in horses. DNA vaccines coding for the structural WNV proteins have also been assessed for veterinary use and have been found to be protective in mice, horses and birds. Live attenuated yellow fever WNV chimeric vaccines have also been successful in animals and are currently undergoing human trials. Additional studies have shown that immunisation with a relatively benign Australian variant of WNV, the Kunjin virus, also provides protective immunity against the virulent North American strain. Levels of efficacy and safety, as well as logistical, economic and environmental issues, must all be carefully considered before vaccine candidates are approved and selected for large-scale manufacture and distribution.
Animals↗
Comparison of the restriction patterns of equine herpesvirus (EHV-1) strains isolated for eight years in France.
The equine herpes viruses strains (EHV) isolated from organs of aborted foetuses or from nasal swabs have been analysed by comparison of their restriction endonucleases patterns using two enzymes, Bam HI and Pst I. The majority of the clinical samples came from the west part of France ("Normandie") after abortions or respiratory disorders. All the viruses isolated were EHV-1 strains whose patterns show considerable homogeneity although some differences can be described. The genomic DNAs of the same twenty strains have been digested by the Pst I enzyme, which induced a great number of restriction fragments. It allows a more precise epidemiological study between strains isolated in the same studs with different Bam HI patterns or between strains with identical Bam HI profiles but with distinct respiratory or abortigenic pathogenicity. No strain isolated from aborted foetuses or nasal swabs presented the vaccinal pattern.
Abortion, Veterinary↗
Quantitative detection of Neospora caninum in bovine aborted fetuses and experimentally infected mice by real-time PCR.
We report the development of a real-time PCR assay for the quantitative detection of Neospora caninum in infected host tissues. The assay uses the double-stranded DNA-binding dye SYBR Green I to continuously monitor product formation. Oligonucleotide primers were designed to amplify a 76-bp DNA fragment corresponding to the Nc5 sequence of N. caninum. A similar method was developed to quantify the 28S rRNA host gene in order to compare the parasite load of different samples and to correct for the presence of potential PCR-inhibiting compounds in the DNA samples. A linear quantitative detection range of 6 logs with a calculated detection limit of 10(-1) tachyzoite per assay was observed with excellent linearity (R(2) = 0.998). Assay specificity was confirmed by using DNA from the closely related parasite Toxoplasma gondii. The applicability of the technique was successfully tested in a variety of host brain tissues: (i) aborted bovine fetuses classified into negative or positive Neospora-infected animals according to the observation of compatible lesions by histopathological study and (ii) experimentally infected BALB/c mice, divided into three groups, inoculated animals with or without compatible lesions and negative controls. All samples were also tested by ITS1 Neospora nested PCR and a high degree of agreement was shown between both PCR techniques (kappa = 0.86). This technique represents a useful quantitative diagnostic tool to be used in the study of the pathogenicity, immunoprophylaxis, and treatment of Neospora infection.
Abortion, Veterinary↗
Early abortion in cattle induced by experimental intrauterine infection with pure cultures of Actinomyces pyogenes.
Actinomyces pyogenes from a case of endometritis was used to study the effects of infection of the bovine embryo between days 27 and 41 of pregnancy. From 10(9) to 10(10) washed organisms were introduced into the uterine lumen of four pregnant cows. Two pregnant cows were inoculated with sterile saline and four pregnant cows were treated with cloprostenol. Embryonic death and abortion followed 29 to 144 hours after the inoculation of the live bacteria. The aborted embryos were macerated or clearly degenerating and yielded profuse pure cultures of A pyogenes. Abortion was accompanied by a sustained increase in uterine tone, opening of the cervix, presence of vaginal pus and a vulval discharge and the persistence of the corpus luteum for at least eight days after abortion. Intrauterine inoculation with saline did not affect pregnancy, but embryonic death, abortion and regression of the corpus luteum occurred 66 to 72 hours after the treatment with cloprostenol. The results suggest that A pyogenes is a primary pathogen and is capable of causing embryonic death and abortion.
Abortion, Veterinary↗
Prevalence of human pathogens in cat and dog fleas in New Zealand.
AIMS: To provide further information on the prevalence of Rickettsia felis, Bartonella hensela, and B. clarridgeiae in cat and dog fleas in New Zealand and their distribution in the country. METHODS: We used PCR and sequencing with primers for the its and pap 31 (for Bartonella spp.), and the gltA and OmpB (for Rickettsia spp.) genes on DNA from fleas collected from dogs and cats presenting to 3 widely separated veterinary practices on the North Island. RESULTS: DNA of R. felis (19%), B. henselae (11%), and B. clarridgeiae (7%) was found in the 114 cat fleas (Ctenocephalides felis) we studied. The DNA of both B. henselae and B. clarridgeiae was found in 3 fleas (from 2 animals); B. clarridgeiae and R. felis in 1 flea; B. henselae and R. felis in 5 fleas (from 3 animals); and R. felis, B. henselae, and B. clarridgeiae in 2 fleas (from 1 animal). No amplicons were obtained from 3 dog fleas (Ctenocephalides canis). CONCLUSIONS: The emerging human pathogens, R. felis, B. henselae, and B. clarridgeiae, are prevalent and widely distributed in cat fleas in the North Island of New Zealand.
Animals↗
Core particles of hepatitis B virus as carrier for foreign epitopes.
To be effective as vaccines, most monomeric proteins and peptides either require chemical coupling to high molecular weight carriers or application together with adjuvants. More recently, recombinant DNA techniques have been used to insert foreign epitopes into proteins with inherent multimerization capacity, such as particle-forming viral capsid or envelope proteins. The core protein of hepatitis B virus (HBcAg), because of its unique structural and immunological properties, has gained widespread interest as a potential antigen carrier. Foreign sequences of up to approximately 40 amino acid residues at the N terminus, 50 or 100 amino acids in the central immunodominant c/e 1 epitope region of HBcAg, and up to 100 or even more residues at the C terminus, did not interfere with particle formation. The humoral immunogenicity of inserted epitopes is determined by the immunogenicity of the peptide itself and its surface exposure, and is influenced by the route of application. The probably flexible and surface-exposed c/e1 region emerged as the most promising insertion site. When applied together with adjuvants approved for human and veterinary use, or even without adjuvants, such chimeric particles induced B and T cell immune responses against the inserted epitopes. In some cases neutralizing antibodies, cytotoxic T cells and protection against challenge with the intact pathogen were demonstrated. Major factors for the potentiated immune response against the foreign epitopes are the multimeric structure of chimeric HBcAg that results in a high epitope density per particle, and the provision of T cell help by the carrier moiety. Beyond its use as subunit vaccine, chimeric HBcAg produced in attenuated Salmonella strains may be applicable as live vaccine.
Epitopes↗
Are we facing a 'post-antibiotic era'?--a review of the literature regarding antimicrobial drug resistance.
Since the introduction of antibiotics in the 1940s, antibiotic resistance has become an increasing problem. Today, multiple-antibiotic resistance is commonly associated with a number of clinically important pathogens and is therefore an important issue in clinical nursing practice. Epidemiological studies identify a number of important factors associated with increases in antimicrobial resistance. These include patterns of antimicrobial use, changes in medical and veterinary care and social practices affecting the transmission of microbes. Bacterial mechanisms of antibiotic resistance and the genetics of resistance-gene transfer are explored, with the intention of developing nurses' knowledge and understanding of control measures.
Anti-Bacterial Agents↗
Mitigation of bioterrorist threats in the 21st century.
There is a raising level of awareness and concern that agriculture and food supplies might serve as targets for bioterrorists. To minimize such threats a number of new initiates are urgently needed: 1. Greater levels of financial commitment from federal, state, and international organizations for research on highly infectious diseases and for emergency response infrastructure; 2. Wellorchestrated emergency response plans based on inter-agency, inter-departmental and stakeholder working groups; 3. Teams prepared for risk assessment and risk communication; 4. Modern systems of animal identification and accurate trace-back for animal movement; 5. Increased biosecurity in intensive production operations; 6. Accurate intelligence as to what pathogens pose the highest risks for economic and social impact; 7. Establishment of new international animal and plant disease research networks and partnerships; 8. Strengthen international disease surveillance networks for early detection and intervention; 9. New generations of rapid diagnostic tests for pathogen detection that are practical for field and diagnostic laboratories; 10. Increased level of commitment to pathogen genomics research for molecular epidemiology and vaccine development; 11. New chemical and immunological intervention strategies to prevent or control disease outbreaks; 12. Increased level of training on exotic and emerging animal diseases in basic veterinary education and through continuing education for veterinarians, state and federal field personnel and laboratory diagnosticians.
Agriculture↗
Vaccine potential of attenuated mutants of Corynebacterium pseudotuberculosis in sheep.
Corynebacterium pseudotuberculosis, a gram-positive facultative intracellular bacterial pathogen, is the etiological agent of the economically important disease caseous lymphadenitis (CLA) in both sheep and goats. Attenuated mutants of C. pseudotuberculosis have the potential to act as novel vaccines against CLA and as veterinary vaccine vectors. In this report, we have assessed the virulence of both aroQ and pld mutants of C. pseudotuberculosis in sheep and concurrently their capacity to act as vaccines against homologous challenge. The results suggest that aroQ mutants of C. pseudotuberculosis are attenuated with regard to both lymph node persistence and vaccination site reactogenicity. Immunologically, aroQ mutants failed to elicit detectable specific gamma interferon (IFN-gamma)-secreting lymphocytes and induced low levels of antibodies to C. pseudotuberculosis culture supernatant antigens. Following subcutaneous vaccination, the immune responses induced by aroQ mutants did not protect sheep from infection with the wild-type strain but did appear to reduce the clinical severity of disease resulting from challenge. Conversely, an attenuated C. pseudotuberculosis strain expressing an enzymatically inactive phospholipase D exotoxin, when used as a vaccine, elicited a protective immune response. Protection appeared to correlate with in vivo persistence of the vaccine strain, the induction of IFN-gamma-secreting lymphocytes, and relatively high levels of antibodies to culture supernatant antigens. The results suggest that aroQ mutants of C. pseudotuberculosis may be overly attenuated for use as a CLA vaccines or as vaccine vectors.
Animals↗
Microbial forensics for natural and intentional incidents of infectious disease involving animals.
Microbial forensics is a relatively new scientific discipline dedicated to analysing microbiological evidence from a crime for attribution purposes. It builds on traditional microbiology and epidemiology but within a legal framework. Important motives for forensic investigations include interdiction of criminals, prosecution of justice, and ideally, deterrence of others from committing similar acts. Forensic capabilities in animal health should focus on building capacity for detection and reporting of increases in infectious disease morbidity and mortality among animals that might reflect a covert release of a pathogen. Suspicion should be raised when epidemiological patterns are different from those expected for the animal population and the pathogen in question. Existing capacities for the detection and reporting of epidemic and even endemic diseases should be an international priority for the prevention of catastrophic losses in animal and potentially in human life. The veterinary community needs to be more aware of the legal requirements related to forensic investigations so that veterinarians will be prepared to handle evidence properly within their own fields.
Animal Diseases↗
West Nile virus in the vertebrate world.
West Nile virus (WNV), an arthropod-borne virus belonging to the family Flaviviridae, had been recognized in Africa, Asia and the south of Europe for many decades. Only recently, it has been associated with an increasing number of outbreaks of encephalitis in humans and equines as well as an increasing number of infections in vertebrates of a wide variety of species. In this article, the data available on the incidence of WNV in vertebrates are reviewed. Moreover, the role of vertebrates in the transmission of WNV, the control of WNV infections in veterinary medicine as well as future perspectives are discussed. A wide variety of vertebrates, including more than 150 bird species and at least 30 other vertebrate species, are susceptible to WNV infection. The outcome of infection depends on the species, the age of the animal, its immune status and the pathogenicity of the WNV isolate. WNV infection of various birds, especially passeriforms, but also of young chickens and domestic geese, results in high-titred viremia that allows arthropod-borne transmission. For other vertebrate species, only lemurs, lake frogs and hamsters develop suitable viremia levels to support arthropod-borne transmission. The role of vertebrates in direct, non-arthropod-borne transmission, such as via virus-contaminated organs, tissues or excretions is less well characterized. Even though direct transmission can occur among vertebrates of several species, data are lacking on the exact amounts of infectious virus needed. Finally, the increased importance of WNV infections has led to the development of killed, live-attenuated, DNA-recombinant and chimeric veterinary vaccines.
Animal Diseases↗
How to modify conditions limiting resistance in bacteria in animals and other reservoirs.
Antimicrobial agents in veterinary medicine are used for three purposes: therapy, prophylaxis, and nutrition. The major public health risk is that selection pressure leads to an increase in the pool of resistance genes. Since 1987, the nutritional use of antimicrobials in Europe has been regulated by a council directive, which demands special investigations into the potential of antimicrobials to increase rates of drug resistance. However, the prophylactic and therapeutic use of antimicrobials has sometimes led to the emergence of resistant bacteria. For example, the selective effect of the prophylactic use of gentamicin and the therapeutic use of quinolones led to the emergence of resistant salmonellae. To prevent the spread of resistant microorganisms from animals to humans, it should be recognized that antibiotics are not suitable as a compensation for poor hygiene standards or for the eradication of a pathogen from a certain environment. They should be used only by doctors or veterinarians.
Animals↗
Pathological and clinical aspects of the diseases caused by Malassezia species.
From veterinary point of view Malassezia pachydermatis has the greatest significance. It has been standing in the focus of interest since the early 1990s, mostly because of the frequency of otitis externa and dermatitis caused by this yeast in dogs. This is the only lipid-independent species in the genus Malassezia. It can be found in very large proportion on the skin of healthy animals, but can be isolated in much greater number from diseased dogs. It often causes illness together with other pathogens (e.g. Staphylococcus intermedius). Some breeds are predisposed. In addition to the treatment of the accidental concurrent diseases, therapy consists of systemic and/or topical antimicrobial treatment. Ketoconazole is used most frequently. Malassezia pachydermatis plays also a role in the skin disorders of other carnivores. It has little zoonotic potential, it can be dangerous to immunocompromised humans. The other Malassezia species have little veterinary importance, although M. sympodialis and M. globosa were isolated from asymptomatic animals (mostly cats) and from mixed infections.
Animals↗
Mystery swine disease in The Netherlands: the isolation of Lelystad virus.
In early 1991, the Dutch pig-industry was struck by the so-called mystery swine disease. Large-scale laboratory investigations were undertaken to search for the etiological agent. We focused on isolating viruses and mycoplasmas, and we tested paired sera of affected sows for antibodies against ten known pig viruses. The mycoplasmas M. hyosynoviae, M. hyopneumoniae, and Acholeplasma laidlawii, and the viruses encephalomyocarditis virus and porcine enterovirus types 2 and 7 were isolated from individual pigs. An unknown agent, however, was isolated from 16 of 20 piglets and from 41 of 63 sows. This agent was characterised as a virus and designated Lelystad virus. No relationship between this virus and other viruses has yet been established. Of 165 sows reportedly afflicted by the disease, 123 (75 per cent) seroconverted to Lelystad virus, whereas less than 10 per cent seroconverted to any of the other virus isolates or to the known viral pathogens. Antibodies directed against Lelystad virus were also found in pigs with mystery swine disease in England, Germany, and in the United States. We conclude that infection with Lelystad virus is the likely cause of mystery swine disease.
Abortion, Veterinary↗