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Anti-enteropathogenic Escherichia coli immunoglobulin Y isolated from eggs laid by immunised Leghorn chickens.

IgY, the egg yolk immunoglobulin, equivalent to the IgG from mammals, has been used in veterinary practice for passive immunisation against bacterial or viral infectious diseases. Enteropathogenic Escherichia coli (EPEC) is the main etiological agent of infantile diarrhoea in Brazil and other developing countries. Our aims were to isolate immunoglobulin IgY from egg yolk laid by EPEC -immunised Leghorn chickens and to study its reactivity to the antigens from this pathogen, including some virulence factors. Leghorn chickens were immunised with a bacterial suspension intramuscularly (three hens) or intravenously (three hens) or with PBS (two hens). Eggs were collected over a period of 17 weeks. IgY isolation procedures were carried out by salt precipitation (ammonium sulphate, in solid form) followed by centrifugations and dialysis. Final preparations were submitted to sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS - PAGE), enzyme-linked immunosorbent assay (ELISA) and immunoblotting. All immunised animals developed good levels of antibodies reactive to whole bacteria or lipopolysaccharide (LPS), in contrast to the control ones. Immunoblottings allowed the recognition of several antigenic fractions of bacterial antigens, some of which had a molecular weight compatible with bacterial virulence factors, confirming the efficacy of the immunisation and the adequacy of the method.

Animals↗

Mycetes and urban areas.

Mycetes are ubiquitous organisms that can cause mycoses in human and animals. The role of animals in the epidemiology of human mycoses in urban areas is multiform, but here will be discuss only two features: A) animals as vectors of mycoses and B) animal substrates as growth factor of pathogenic fungi. A) Animals as vectors of mycoses: this role is important as zoofilic dermatophytes are very important agents of zoonosis; the urban dermatophytozoonoses are prevalent caused by Microsporum canis which is prevalent in cats and dogs. Cats are often asymptomatic carriers. The pattern of human dermatomycoses has changed in Italy during the past century: at the beginning of the century anthropophilic fungi were prevalent while at present the zoophilic fungi are the most important causes. B) Animal substrata as growth factor of pathogenic fungi: soil "animalization" (i.e., the addition of such debris as hair, skin scales, dropping and other organic matters) creates an optimal substratum for the growth and the multiplication of geophilic or saprophyitic fungi, such as Microsporum gypseum and Cryptococcus neoformans. The present human lifestyle, which favours a an overpopulation of birds, wild animals, domestic mammals and sinanthropic together with man in crowded areas seems to favour the formation of environments adapted to the abundant growth of some pathogenic fungi with consequent infection for man and animals. Finally, an environment heavily populated by fungi can cause allergic pulmonary reactions as well as reactions in other organs and tissues. The control of human and animal fungi, and the efficient use of a monitoring system require ample knowledge of mycological problems both in human and veterinary medicine and of efficient laboratories capable of resolving the needs of both disciplines. Close collaboration between veterinarians, doctors and mycologists is necessary in order to resolve health problems linked to mycosis.

Animals↗

Multidrug resistance in parasites: ABC transporters, P-glycoproteins and molecular modelling.

Parasitic diseases, caused by protozoa, helminths and arthropods, rank among the most important problems in human and veterinary medicine, and in agriculture, leading to debilitating sicknesses and loss of life. In the absence of vaccines and with the general failure of vector eradication programs, drugs are the main line of defence, but the newest drugs are being tracked by the emergence of resistance in parasites, sharing ominous parallels with multidrug resistance in bacterial pathogens. Any of a number of mechanisms will elicit a drug resistance phenotype in parasites, including: active efflux, reduced uptake, target modification, drug modification, drug sequestration, by-pass shunting, or substrate competition. The role of ABC transporters in parasitic multidrug resistance mechanisms is being subjected to more scrutiny, due in part to the established roles of certain ABC transporters in human diseases, and also to an increasing portfolio of ABC transporters from parasite genome sequencing projects. For example, over 100 ABC transporters have been identified in the Escherichia coli genome, but to date only about 65 in all parasitic genomes. Long established laboratory investigations are now being assisted by molecular biology, bioinformatics, and computational modelling, and it is in these areas that the role of ABC transporters in parasitic multidrug resistance mechanisms may be defined and put in perspective with that of other proteins. We discuss ABC transporters in parasites, and conclude with an example of molecular modelling that identifies a new interaction between the structural domains of a parasite P-glycoprotein.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Clinical pharmacokinetics of parenterally administered danofloxacin in cattle.

Danofloxacin is a new fluoroquinolone antibacterial, developed specifically for veterinary use. Its in vitro activity and pharmacokinetic properties have been investigated to assess its potential for use in the therapy of respiratory disease in cattle. The minimum inhibitory concentration of danofloxacin against 90% (MIC90) of contemporary European and North American field isolates of Pasteurella haemolytica, Pasteurella multocida and Haemophilus somnus, the most important bacterial respiratory pathogens of cattle, was 0.125 micrograms/ml. The plasma and lung kinetics of danofloxacin following parenteral administration of 1.25 mg/kg were evaluated in two studies. Danofloxacin was rapidly absorbed following intramuscular and subcutaneous injection and bioavailability was virtually complete (101% and 94% respectively). Plasma concentration profiles of danofloxacin were similar for intramuscular and subcutaneous routes with no significant differences in the area under the plasma concentration-time curves (AUC) following one, three or five consecutive daily doses, although slightly higher peak plasma concentrations were achieved by the intramuscular route. Following intramuscular administration, the mean peak lung concentration of danofloxacin was 4.1 times greater than that of plasma. Similarly, the AUC for lung tissue was 3.7 times greater than that for plasma. These data indicate that danofloxacin should be particularly appropriate for the therapy of bacterial respiratory disease in cattle.

Animals↗

Antimicrobial resistance and class 1 integrons in pathogenic Escherichia coli from dairy farms.

The goal of this study was to assess the prevalence of antimicrobial resistance and class 1 integrons, including integron-associated genes, in 24 Escherichia coli isolates from dairy farms. Escherichia coli isolates (n = 14) from dairy cows with mastitis (ECDM), Shiga toxin-producing (STEC) O157:H7 from cull dairy cow fecal samples (n = 9) and bulk tank milk (n = 1) were evaluated for sensitivity to 19 antimicrobial agents used commonly in human and/or veterinary medicine. Multiplex PCR was used to determine presence of genes associated with class 1 integrons (intI1, qacEDelta1, and sulI1). Class 1 integrons were found only in eight of 10 isolates (one STEC O157:H7 and seven ECDM) that demonstrated antimicrobial resistance, and seven of these were resistant to two or more antimicrobial agents. Eight of 10 STEC O157:H7 and six of 14 ECDM were susceptible to all commonly used antibiotics. Five ECDM demonstrated multiple resistances to four or more antibiotics. Most of the 24 isolates examined exhibited resistance against sulfamethoxazole, followed by streptomycin and tetracycline. STEC O157:H7 strains had less prevalence of antibiotic resistance and integron carriage than ECDM. The multiplex PCR method developed for detection of intI1, qacEDelta1, and sulI1 can be used routinely for monitoring presence of these genes. Class 1 integrons were found in eight of 10 E. coli strains that demonstrated antimicrobial resistance; seven of these were resistant to two or more antibiotics. It appears that integrons played a role in the incidence of antimicrobial resistance of the strains used in this study.

Animals↗

Backgrounding beef cattle.

Stocker operations differ significantly in management techniques and health programs. Critical control points should be identified for each farm and used to ascertain the biggest areas for potential improvement so as to allow proper allocation of resources. A customized wellness program couples medicine and management to minimize the negative impact of disease. A complete program involves animal selection, disease prevention, and management. Calves should be evaluated and classified into appropriate risk categories regarding treatment, management, and performance expectations. Disease prevention strategies should involve increasing immune responses while decreasing pathogen exposure and stress in cattle. Disease management includes the methods used to minimize the impact of illness on the population

Animal Husbandry↗

Pattern recognition receptors in companion and farm animals - the key to unlocking the door to animal disease?

The innate immune system is essential for host defence and is responsible for early detection of potentially pathogenic microorganisms. Upon recognition of microbes by innate immune cells, such as macrophages and dendritic cells, diverse signalling pathways are activated that combine to define inflammatory responses that direct sterilisation of the threat and/or orchestrate development of the adaptive immune response. Innate immune signalling must be carefully controlled and regulation comes in part from interactions between activating and inhibiting signalling receptors. In recent years, an increasing number of pattern recognition receptors (PRRs), including C-type lectin receptors and Toll-like receptors (TLRs), has been described that participate in innate recognition of microbes, especially through the so called pathogen-associated molecular patterns (PAMPs). Recent studies demonstrate strong interactions between signalling through these receptors. Whereas useful models to study these receptors in great detail in the murine and human system are now emerging, relatively little is known regarding these receptors in companion and farm animals. In this review, current knowledge regarding these receptors in species of veterinary relevance is summarised.

Animals↗

Management of avian flock emergencies.

Managing avian flock emergencies requires a thorough history, physical examination, and record review before instituting appropriate therapy. Although the general thought processes are similar between approaching a sick individual bird and a "diseased" aviary, the actual steps are different. Infectious diseases are often a component of aviary emergencies but are rarely the primary cause of flock "disease." Managing flock emergencies typically goes beyond addressing individual pathogens. Recognizing, evaluating, and treating flock emergencies are discussed with the intent to provide a better understanding of flock health medicine principles.

Animal Husbandry↗

Pharmacological approaches towards rationalizing the use of endoparasitic drugs in small animals.

Parasitic diseases are an important health concern to small animal veterinarians worldwide, and their zoonotic potential is also of relevance to human medicine. The treatment and control of such conditions relies heavily on pharmaceutical intervention using a range of antiparasitic drugs and/or their biologically active metabolites. Broad spectrum agents have been produced, although narrow and even monospecific drugs are used in some situations. Their efficacy may depend on dosage, the target pathogen(s), the host species and/or the site of infection. Optimal use of antiparasitics requires a detailed consideration of the pharmacokinetic and pharmacodynamic properties of the drugs in specific clinical contexts. This review summarizes the present status of knowledge on the metabolism, and physicochemical and pharmacological properties of the major antiparasitic drugs currently used in small animal veterinary practice. In addition, data relevant to therapeutic dosage, efficacy and clinical indication/contraindication, particularly in relation to combination drug therapy, are included.

Animals↗

Characterization of avian influenza virus isolates submitted to the National Centre for Foreign Animal Disease between 1997 and 2001.

The National Centre for Foreign Animal Disease (NCFAD) in Winnipeg, Manitoba, is the Canadian Food Inspection Agency's (CFIA) newest high biocontainment laboratory. One of the functions of the NCFAD is to serve as a national reference laboratory for avian influenza. Between 1997 and 2001, 15 avian influenza virus isolates were characterized. These isolates originated from domestic poultry, imported caged birds held in quarantine, and wild birds. Diagnostic specimens were submitted to the NCFAD by CFIA field veterinarians, provincial veterinary diagnostic laboratories, and veterinary colleges. Characterization of isolates included the determination of H and N subtypes: H1, H6, H7, and H10 subtypes were isolated from domestic poultry; H3, H4, and three H13 viruses were isolated from water fowl, and six H3 viruses were isolated from caged birds being held in import quarantine. Selected isolates were characterized with respect to their pathogenic potential by intravenous inoculation of 4-to-6-wk-old chickens. A molecular-based protocol was used to assess the pathogenicity of one H7 isolate. During this period, work was also carried out toward validating our molecular pathotyping protocol for avian influenza viruses with H5 and H7 hemagglutinin subtypes.

Amino Acid Sequence↗

Reproductive failure in mink and ferrets after intravenous or oral inoculation of Campylobacter jejuni.

Four pregnant mink and seven pregnant ferrets, including five with previous exposure and specific antibody, were injected intravenously with 10(8)-10(10) colony-forming units of Campylobacter jejuni. All 11 pregnancies failed 1-16 days after infection, with results ranging from fetal resorption to expulsion of dead or premature living kits. In every case, uterine contents (placenta, uterine fluid and/or kits) were culture-positive for C. jejuni. Three pregnant mink and nine pregnant ferrets, including four with previous exposure and antibody, were fed 10(9)-10(11) C. jejuni. Two of the mink aborted; kits of all three were culture-positive, but those of one female survived. Seven of the nine ferrets aborted, with two having culture-positive uterine contents. None of 28 uninfected ferret control pregnancies ended in abortion. The most prominent histological feature observed was severe placentitis, which appears to be a more likely cause of Campylobacter-induced abortion than direct pathogenic effects on infected kits. These results suggest that infection of mink or ferrets with C. jejuni during pregnancy poses a serious risk of reproductive failure, even for previously exposed females.

Abortion, Veterinary↗

Recent advances in veterinary vaccine adjuvants.

Next generation veterinary vaccines are going to mainly comprise of either subunit or inactivated bacteria/viruses. These vaccines would require optimal adjuvants and delivery systems to accord long-term protection from infectious diseases in animals. There is an urgent need for the development of new and improved veterinary and human vaccine adjuvants. Adjuvants can be broadly divided into two classes, based on their principal mechanisms of action: vaccine delivery systems and 'immunostimulatory adjuvants'. Vaccine delivery systems are generally particulate e.g. emulsions, microparticles, ISCOMS and liposomes, and mainly function to target associated antigens into antigen presenting cells (APC). In contrast, immunostimulatory adjuvants are predominantly derived from pathogens and often represent pathogen associated molecular patterns, e.g. LPS, MPL and CpG DNA, which activate cells of the innate immune system. Recent progress in innate immunity is beginning to yield insight into the initiation of immune responses and the ways in which immunostimulatory adjuvants might enhance this process in animals and humans alike.

Adjuvants, Immunologic↗

Developmentally regulated biosynthesis of carbohydrate and storage polysaccharide during differentiation and tissue cyst formation in Toxoplasma gondii.

Toxoplasma gondii belongs to the Apicomplexa phylum, which comprises protozoan parasites of medical and veterinary significance, responsible for a wide variety of diseases in human and animals, including malaria, toxoplasmosis, coccidiosis and cryptosporidiosis. During infection in the intermediate host, T. gondii undergoes stage conversion between the rapidly replicating tachyzoite that is responsible for acute toxoplasmosis and the dormant or slowly dividing encysted bradyzoite. The tachyzoite-bradyzoite interconversion is central to the pathogenic process and is associated with the life-threatening recrudescence of infection observed in immunocompromised patients such as those suffering from AIDS. In chronic infections, the bradyzoites are located within tissue cysts found predominantly in brain and muscles. The tissue cyst is enclosed by a wall containing specific lectin binding sugars while the bradyzoites have accumulated large amounts of the storage polysaccharide of glucose, amylopectin. Our recent findings have identified several genes and proteins associated with amylopectin synthesis or degradation and glucose metabolism, including different isoforms of certain glycolytic enzymes, which are stage-specifically expressed during tachyzoite-bradyzoite interconversion. Here, we will discuss how the genes and enzymes involved in carbohydrate metabolisms are used as molecular and biochemical tools for the elucidation of molecular mechanisms controlling T. gondii stage interconversion and cyst formation.

Amylopectin↗

A rare genotype of Cryptococcus gattii caused the cryptococcosis outbreak on Vancouver Island (British Columbia, Canada).

Cryptococcus gattii causes life-threatening infection of the pulmonary and central nervous systems in hosts with normal immunity and traditionally has been considered to be restricted geographically to tropical and subtropical climates. The recent outbreak of C. gattii in the temperate climate of Vancouver Island, BC, Canada, led to a collaborative investigation. The objectives of the current study were to ascertain the environmental source of the outbreak infections, survey the molecular types of the outbreak and environmental cryptococcal isolates, and determine the extent of genetic diversity among the isolates. PCR-fingerprinting and amplified fragment length polymorphism (AFLP) were used to examine the genotypes, and mating assays were performed to determine the mating type of the isolates. All outbreak and environmental isolates belonged to C. gattii. Concordant results were obtained by using PCR-fingerprinting and AFLP analysis. The vast majority of clinical and veterinary infections were caused by isolates of the molecular type VGII/AFLP6, but two were caused by molecular type VGI/AFLP4. All environmental isolates belonged to molecular type VGII/AFLP6. Two or three subtypes were observed within VGII/AFLP6 among outbreak and environmental isolates. All mating-competent isolates were of the alpha-mating type. The emergence of this usually tropical pathogen on Vancouver Island highlights the changing distribution of this genotype and emphasizes the importance of an ongoing collaborative effort to monitor the global epidemiology of this yeast.

Adolescent↗

[Ectoparasites of animals: methods of ecological, biological, genetic and mechanical control].

The use of insecticides is still the basic procedure for controlling most ectoparasites, but various methods are being developed to act in addition to, or in synergy with these products, so as to enhance the efficacy and reduce the adverse effects of insecticides, by contributing to ecologically acceptable strategies. These methods are classified as ecological control (modification of the environment of the parasite), biological control (predation, parasitism, action of pathogens, etc.), genetic control (release of sterile males, hybridisation, genetic manipulations) and mechanical control (insect traps, use of repellents). The application of such methods depends on the biological and ecological characteristics of the ectoparasite, and they may act directly or indirectly, affecting mortality and/or reproduction. The authors review the principal methods applicable to major groups of ectoparasites of veterinary interest. Non-chemical methods are the subject of wide-ranging and promising research, particularly in view of recent developments in biotechnology.

Animals↗

[Pathways and procedures of mycological diagnosis in veterinary laboratories. A diagnostic approach].

With the increasing fungal affections in humans and animals, the veterinary laboratories will have to reckon with more fungal specimens. A reliable and rapid identification of the etiologic agents will be required. The procedure with mould cultures is described in the general part. In the specific part, the general steps to differenciate medical important moulds (dermatophyes, zygomycetes, dematiaceous fungi, Aspergillus, etc.) are delineated. The identification of some fungal pathogens is possible by the means of the schematic representations.

Animals↗

Silage and animal health.

The process of preserving crops by fermentation in silos is under the control of the farmer to a much lesser degree compared to the level of control by the manufacturer over the production of other fermented foods, such as cheese and yoghurt. Additives designed to direct the extent and pattern of the fermentation are relatively unpopular in most countries, and their use is not guaranteed to remove the risk of undesirable components in silage. Hazards to animal health associated with silage fall into three categories: (1) undesirable micro-organisms e.g. Listeria, enterobacteria, clostridia and moulds; (2) undesirable chemicals, e.g. mycotoxins, and (3) excess acidity and other metabolic disorders. In some regions of Europe, the production of silage is discouraged or prohibited because of the risk of undesirable microbes. The princIpal risk in these areas is that of the secondary fermentation of cheese made from milk contaminated by bacterial spores, rather than a direct hazard of contaminated silage to animal health. With the possible exception of high dry matter silage conserved in large bales, respiratory hazards to animals from moulds and their spores generally are less from silage than hay. Mycotoxins and phytoestrogens may survive the ensiling period and constitute risks to animal health. Relatively little is known about the epidemiology of diseases that may be linked to undesirable chemicals and excess acidity in silage. Therefore, research is needed to define epidemiologically and mechanistically the risks to animal health and to the human food chain from silages contaminated with pathogenic bacteria and mycotoxins, and to understand more completely the relationships between the physical and chemical compositions of silage and metabolic disorders in animals.

Animal Diseases↗