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Subtracted restriction fingerprinting--a new typing technique using magnetic capture of tagged restriction fragments.

Molecular typing of bacterial pathogens is an important issue in the epidemiological analysis of emerging infections in humans and animals. Numerous methods have been developed for and applied to a wide variety of bacteria of medical, veterinary and zoonotic importance. The present minireview provides a description of a new typing approach designated subtracted restriction fingerprinting (SRF), its use for typing of Salmonella isolates and a comparison with the most widely used typing techniques for these bacteria. SRF is based on double restriction endonuclease digestion of whole cell DNA, followed by a fill-in reaction with specifically tagged nucleotides and subtractive capture of selected restriction fragments. This results in a reduced number of fragments optimal for separation in standard agarose gels.

Animals↗

Are ducks contributing to the endemicity of highly pathogenic H5N1 influenza virus in Asia?

Wild waterfowl are the natural reservoir of all influenza A viruses, and these viruses are usually nonpathogenic in these birds. However, since late 2002, H5N1 outbreaks in Asia have resulted in mortality among waterfowl in recreational parks, domestic flocks, and wild migratory birds. The evolutionary stasis between influenza virus and its natural host may have been disrupted, prompting us to ask whether waterfowl are resistant to H5N1 influenza virus disease and whether they can still act as a reservoir for these viruses. To better understand the biology of H5N1 viruses in ducks and attempt to answer this question, we inoculated juvenile mallards with 23 different H5N1 influenza viruses isolated in Asia between 2003 and 2004. All virus isolates replicated efficiently in inoculated ducks, and 22 were transmitted to susceptible contacts. Viruses replicated to higher levels in the trachea than in the cloaca of both inoculated and contact birds, suggesting that the digestive tract is not the main site of H5N1 influenza virus replication in ducks and that the fecal-oral route may no longer be the main transmission path. The virus isolates' pathogenicities varied from completely nonpathogenic to highly lethal and were positively correlated with tracheal virus titers. Nevertheless, the eight virus isolates that were nonpathogenic in ducks replicated and transmitted efficiently to naïve contacts, suggesting that highly pathogenic H5N1 viruses causing minimal signs of disease in ducks can propagate silently and efficiently among domestic and wild ducks in Asia and that they represent a serious threat to human and veterinary public health.

Animals↗

Clinical pharmacology of antimicrobial use in humans and animals.

Veterinary public health is a frontier in the fight against human disease, charged to control and eradicate zoonotic diseases that are naturally transmitted between vertebrate animals and man. Currently there is a need for clinical pharmacologists and all health care givers to limit the development of bacterial resistance in humans to contain the increased health care expenditures related to morbidity and mortality associated with the use of antimicrobials. The development of resistance predates the use of antibiotics and will always be a problem to the successful treatment of patients. Ongoing discussion debates the extent to which antibiotic use in animals contributes to the development of antibiotic resistance in humans. The veterinary use ofantibiotics as antimicrobial growth promoters is thought to influence the prevalence of resistance in animal bacteria and to be a risk factor for the emergence of antibiotic resistance in human pathogens. Transfer of antibiotic resistant bacteria from animals to humans may occur via contact, including occupational exposure and via the food chain. Resistance genes may transferfrom bacteria of animals to human pathogens in the intestinal flora of humans. Prevention of the development of resistance in humans necessitates good animal husbandry and hygienic measures to prevent cross contamination and a decrease in the use of antibiotics. Appropriate use of antibiotics for food animals will preserve the long-term efficacy of existing antibiotics, support animal health and welfare, and limit the risk of transfer of antibiotic resistance to humans. Investigators must also develop new antimicrobial agents. Poole (J Pharmacy Pharmacol 2001;53:283) recommends targeting the three predominate mechanisms of development of resistance by antimicrobials (i.e., antibiotic inactivation, target site modification, and altered uptake via restricted entry and/or enhanced efflux) to specifically complement the development of novel agents with novel bacterial targets. Bacterial resistance and its selection may be evaluated by comparing the relationship to antibiotic pharmacokinetic (PK) values obtained from serum concentrations and organism MICs (minimum inhibitory concentrations; concentration-dependent killing) to reveal culture and sensitivity tests in patients. Pharmacodynamic (PD) models may be developed to identify factors associated with the probability that bacterial resistance will develop. Thomas et al (Antimicrobial Agents Chemotherapy 1998;42:521) used this combined approach of PK/PD and MICs to examine data retrospectively. The role of clinical pharmacology is to work with PK/PD models such as these to determine the best use of antibiotics in humans to minimize the development of resistance. The role of any regulatory body responsible for the protection of the public health and food safety for consumers is to assess risk and to then communicate and manage the risk. Scientific uncertainty must be interpreted to propose sound policy options. The conversion of sound science into an appropriate regulatory policy to protect the public health is most important.

Animals↗

History and evolution of HPAI viruses in southeast Asia.

Highly pathogenic avian influenza (HPAI) has been recognized as a serious viral disease of poultry since 1878. The number of outbreaks of this disease globally has increased in the past 10 years culminating in 2004 with the unprecedented outbreak of H5N1 HPAI involving nine countries in East and South East Asia. Apart from the geographical extent of this outbreak and apparent rapid spread, this epidemic has a number of unique features, among which is the carriage of highly pathogenic AI viruses by asymptomatic domestic waterfowl. When this disease first emerged it was recognized almost simultaneously in a number of countries for the first time. This created considerable concern among both veterinary and public health authorities especially as the virus was also shown to cause fatal disease in humans. This article brings together a range of information on H5N1 HPAI viruses in Asia that were collected by FAO during the past year through field projects and explores possible reasons for the emergence of the disease in late 2003 and early 2004. Key epidemiological features of the disease in different Asian countries are described in an attempt to look for, and where possible, explain similarities and differences. This includes assessment of factors that could have contributed to the spread of the disease. Molecular aspects of the viruses are examined to assess relationships between isolates from different locations and times so as to gain insights into the origins of viruses in various countries. It is apparent that the coincidence and grouping of the reports declaring the outbreaks of HPAI did not truly reflect the time course of disease emergence, which was widespread well before the outbreak. The factors that could have led to a change from infection to emergence of widespread disease in 2003-2004 are discussed. There are still some questions that remain unanswered regarding the origins of the 2004 outbreak. This article does not provide answers to all of these, but brings together what is currently known about these outbreaks and the viruses that have caused them.

Agriculture↗

Disease risks associated with wildlife translocation projects.

Translocation is defined as the movement of living organisms from one area for free release in another. Throughout the world, increasing numbers of native and exotic species are translocated every year. Most of these movements involve native mammals, birds and fish, and are made by private and national wildlife agencies to augment existing populations, usually for sporting purposes. The translocation of endangered species, often to reintroduce them into a part of the historical range from which they have been extirpated, has also become an important conservation technique. The success of potentially expensive, high-profile wildlife translocation projects depends to a large extent on the care with which wildlife biologists and their veterinary advisers evaluate the suitability of the chosen release site, and on the ability of the translocated animals to colonise the area. The veterinary aspects of reintroduction projects are proving to be of extreme importance. There are already instances of inadequate disease risk assessment resulting in expensive failures and, worse still, the introduction of destructive pathogens into naive resident wildlife populations. In this paper, some of the disease risks attending wildlife translocation projects are described and suggestions are made for the development of systematic procedures to reduce these risks both at the source of the founder animals and at the proposed release site.

Animal Diseases↗

[Borrelia burgdorferi from Ixodes ricinus ticks on Ameland].

Ixodes ricinus ticks collected from the North Sea Island of Ameland, the Netherlands, were screened for the presence of Borrelia spirochaetes. The present authors used IFAT on tick gut preparations and identified Borrelia in 31 per cent of the adult ticks and 16 per cent of the nymphs collected from the dunes and 45 per cent of the nymphs collected from the forests. In vitro cultivation of Borrelia sp. isolated from the adult ticks and its subsequent recognition by B. burgdorferi-specific monoclonal antibodies in western blots, verified that this pathogen is identical with B. burgdorferi, the causative agent of Lyme Disease. Further research is required to gain an insight into the epidemiology of the disease and to assess its medical and veterinary significance in the Netherlands.

Animals↗

Pharmacokinetic/pharmacodynamic relationships of antimicrobial drugs used in veterinary medicine.

The rise in incidence of antimicrobial resistance, consumer demands and improved understanding of antimicrobial action has encouraged international agencies to review the use of antimicrobial drugs. More detailed understanding of relationships between the pharmacokinetics (PK) of antimicrobial drugs in target animal species and their action on target pathogens [pharmacodynamics (PD)] has led to greater sophistication in design of dosage schedules which improve the activity and reduce the selection pressure for resistance in antimicrobial therapy. This, in turn, may be informative in the pharmaceutical development of antimicrobial drugs and in their selection and clinical utility. PK/PD relationships between area under the concentration time curve from zero to 24 h (AUC(0-24)) and minimum inhibitory concentration (MIC), maximum plasma concentration (C(max)) and MIC and time during which plasma concentrations exceed the MIC have been particularly useful in optimizing efficacy and minimizing resistance. Antimicrobial drugs have been classified as concentration-dependent where increasing concentrations at the locus of infection improve bacterial kill, or time-dependent where exceeding the MIC for a prolonged percentage of the inter-dosing interval correlates with improved efficacy. For the latter group increasing the absolute concentration obtained above a threshold does not improve efficacy. The PK/PD relationship for each group of antimicrobial drugs is 'bug and drug' specific, although ratios of 125 for AUC(0-24):MIC and 10 for C(max):MIC have been recommended to achieve high efficacy for concentration-dependent antimicrobial drugs, and exceeding MIC by 1-5 multiples for between 40 and 100% of the inter-dosing interval is appropriate for most time-dependent agents. Fluoroquinolones, aminoglycosides and metronidazole are concentration-dependent and beta-lactams, macrolides, lincosamides and glycopeptides are time-dependent. For drugs of other classes there is limited and conflicting information on their classification. Resistance selection may be reduced for concentration-dependent antimicrobials by achieving an AUC(0-24):MIC ratio of greater than 100 or a C(max):MIC ratio of greater than 8. The relationships between time greater than MIC and resistance selection for time-dependent antimicrobials have not been well characterized.

Animals↗

Johne's disease eradication and control: regulatory implications.

The time has come for the livestock industry and the veterinary profession to take Johne's disease seriously in the United States. The continued spread of the disease with accompanying economic hardship to infected herds and the possibility that M. paratuberculosis may be a food-borne pathogen makes inaction a costly proposition. Efforts have started at the national level with NJWG to have guidelines written for a national control program for Johne's disease. A national control program is desirable because it would provide uniformity to control efforts. Veterinarians and livestock owners should be aware of the effort and let their views be known. In addition, national program guidelines already have been written (see the appendix) to certify test-negative herds for paratuberculosis. States that do not offer the certification program should consider it. A list of test-negative herds for Johne's disease that livestock owners could use to buy low risk animals would be a significant step forward in our efforts to control Johne's disease. Federal regulations concerning Johne's disease are outdated and should be changed to facilitate participation by the livestock industry in a Johne's disease control and eradication program.

Animals↗

Equine infectious respiratory disease.

During the past 20 years the equine population of Great Britain and Ireland has increased with the result that the practising veterinary surgeon is more frequently called upon to advise on equine problems. A significant portion of this advice is concerned with the examination of horses showing signs of this advice is concerned with the examination of horses showing signs of respiratory disease. Numerous pathogens, which include viruses, bacteria, parasites and moulds invade the respiratory tract causing similar signs of illness. It is therefore difficult to provide an aetiological diagnosis based on a clinical examination. Field studies supported by laboratory investigations have established that influenza and herpes viruses are frequently responsible for outbreaks of disease. Epidemiological studies suggest that other factors including the immune state of the equine population influence the distribution and severity of respiratory disease. The aetiology, diagnosis, treatment and control of equine infectious respiratory disease are discussed below.

Abortion, Veterinary↗

Susceptibility testing for bovine respiratory and enteric disease.

The interpretation of susceptibility results for antimicrobials with NCCLS-approved veterinary-specific breakpoints and where the methods also were NCCLS-approved are well established. When these same breakpoints are applied to other applications, however, the interpretation is not so clear. In these cases, a finding of S based on serial-dilution breakpoints puts the isolate in a defined population of bacteria with an MIC equal to or below the S breakpoint. An R result, in these cases, indicates that the organism may have an MIC equal to or greater (with no limits) than the R breakpoint. Extended-dilution testing yields more specific information about the isolate MIC. The relationship of disk-diffusion zone diameters to serial-dilution MICs is correlated on the basis of specific bacterial populations. When disk-diffusion results are interpreted for isolates other than those used for interpretive criteria development, the clinician is left wondering if the zone-diameter results now have a different relationship to serial-dilution results. Furthermore, the question of predictive value of the serial-dilution break-points still remains. The veterinary clinician should be aware of the differences in susceptibility testing predictive value for different applications. When approved veterinary-specific interpretive criteria are not available, then it is appropriate to keep records of clinical response related to susceptibility testing results for common therapies. Advice should be sought on the relationship of pathogen MICs to pharmacokinetic-pharmacodynamic parameters in these situations.

Animals↗

Field studies show associations between pulsator characteristics and udder health.

Four different field studies including 64, 140, 850 and 180 herds were used to investigate the association between pulsator characteristics and udder health. The results indicate that the best udder health is found in herds with a d phase > 330 ms and a pulsation rate > 55 min-1. A d phase of < 250 ms was associated with significant elevation in bulk milk somatic cell count, a higher incidence of acute clinical mastitis and a higher frequency of cows having mastitis associated with major pathogens (mainly Staphylococcus aureus) as compared with a d phase of > 331 ms. There was also a significant positive association between a shorter d phase and a high incidence of teat lesions needing veterinary treatment. Herds having no such teat lesions had pulsators with significantly longer d phase than herds with teat lesions (300 ms v. 288 ms). These results strongly suggest that in high-line milking machines the d phase should not be as short as 250 ms and should preferably be approximately 300 ms, and the pulsation rate should be at least > 55 cycles min-1. There was a strong relationship between the effect of d phase and the rate of pulsation.

Animals↗

Antimicrobial resistance in livestock.

Antimicrobial resistance may become a major problem in veterinary medicine as a consequence of the intensive use and misuse of antimicrobial drugs. Related problems are now arising in human medicine, such as the appearance of multi-resistant food-borne pathogens. Product characteristics, dose, treatment interval and duration of treatment influence the selection pressure for antimicrobial drug resistance. There are theoretical, experimental and clinical indications that the emergence of de novo resistance in a pathogenic population can be prevented by minimizing the time that suboptimal drug levels are present in the infected tissue compartment. Until recently, attention has been focused on target pathogens. However, it should be kept in mind that when antimicrobial drugs are used in an individual, resistance selection mainly affects the normal body flora. In the long term, this is at least equally important as resistance selection in the target pathogens, as the horizontal transfer of resistance genes converts almost all pathogenic bacteria into potential recipients for antimicrobial resistance. Other factors contributing to the epidemiology of antimicrobial resistance are the localization and size of the microbial population, and the age, immunity and contact intensity of the host. In livestock, dynamic herd-related resistance patterns have been observed in different animal species.

Animals↗

A review of the use of teicoplanin in haematological malignancy.

Factors determining the change in patterns of antibiotic use in patients with haematological malignancies include the growing numbers of infections caused by Gram-positive pathogens, the increasing reliance on indwelling catheters, and strategic movement away from inpatient therapy towards day-case and non-inpatient therapy. The suitability of teicoplanin in this context is examined. The indications for teicoplanin use are now established as: early use in clinically infected patients; as a second-line agent for sensitive pathogens; and in primary antibiotic failures. Teicoplanin should not be used as empirical therapy in fevers of unknown origin; neither should teicoplanin be used as prophylaxis (the situations of infection in chemotherapy patients or for Hickman catheter insertion in particular are discussed). Comparative studies versus vancomycin have shown teicoplanin to be equally effective and to be well tolerated. Teicoplanin's good safety profile and ease of use make it particularly useful for non-inpatient therapy. Problems are posed for both glycopeptides by specific resistant pathogens, although these pathogens are currently not common. Haematologists must be aware of, and guard against, abuse of the glycopeptides in their own and other fields, including oral gut decontamination, the treatment of Clostridium difficile and veterinary use.

Anti-Bacterial Agents↗

Equine neonatal septicaemia: 24 cases.

Equine neonatal septicaemia was confirmed in 24 foals hospitalised at the Rural Veterinary Centre between 1989 and 1992 with suspected septicaemia. Septicaemia was confirmed by culture of bacteria from blood of live foals and tissues obtained at necropsy of foals that died or were euthanased. Pathogenic bacteria isolated were predominantly Enterobacteriaceae (including Escherichia coli and Salmonella serovars) and Actinobacillus equuli. Clinical manifestations of septicaemia included signs of depression, dehydration, abnormalities in body temperature and manifestations of localised infection including diarrhoea, pneumonia, and septic arthritis. Most common haematological abnormalities were neutropenia and increase of circulating band neutrophils. Survival rate of foals with confirmed septicaemia was 70.8%. Survival was found to be less likely in the presence of pneumonia, severe signs of depression, marked haematological changes or septic arthritis at the time of admission. Seven foals were confirmed to have septic arthritis without concurrent septicaemia. Of these, 4 had multiple joint involvement. Bacteria isolated from infected joints were predominantly Salmonella serovars. Four foals with septic arthritis failed to survive, due to multiple joint infection, which was unresponsive to treatment. The clinical and haematological abnormalities present in foals with confirmed septicaemia and septic arthritis were consistent with those observed in other studies. The bacterial isolates from foals with confirmed septicaemia were similar to those isolated in other studies. In contrast, the bacteria isolated from foals with septic arthritis without concurrent septicaemia were different from other studies.

Animals↗

Molecular techniques to detect and identify protozoan parasites in the environment.

The environmental route of transmission is important for many protozoan and helminth parasites, with water, soil and food being particularly significant. Both the potential for producing large numbers of transmissive stages and their environmental robustness pose persistent threats to public and veterinary health. The introduction of molecular techniques, in particular those based on the amplification of nucleic acids, has provided researchers with highly sensitive and specific assays for the detection and identification of these pathogens. The application of these techniques to clinical, environmental, and food samples is instrumental for a thorough understanding of the epidemiology of the infection and for the implementation of control measures. Here, the advantages and drawbacks of some molecular techniques (Polymerase Chain Reaction--PCR; Reverse-Transcriptase PCR--RT-PCR; Real-time PCR--qPCR; Nucleic Acid Sequence-Based Amplification--NASBA) will be briefly reviewed. Some application of these techniques will be illustrated with reference to two important and widespread human parasites, the apicomplexan Cryptosporidium and the flagellate Giardia.

Animals↗

Current status of antibiotic resistance in animal production.

It is generally accepted that the more antibiotics we use, the faster bacteria will develop resistance. Further it has been more or less accepted that once an antibiotic is withdrawn from the clinic, the resistance genes will eventually disappear, [table: see text] since they will no more be of any survival value for the bacterial cell. However, recent research has shown that after a long time period of exposure to antibiotics, certain bacterial species may adapt to this environment in such a way that they keep their resistance genes stably also after the removal of antibiotics. Thus, there is reason to believe that once resistance has developed it will not even in the long term be eradicated. What then can we do not to increase further the already high level of antibiotic-resistant bacteria in animals? We should of course encourage a prudent use of these valuable drugs. In Sweden antibiotics are not used for growth promoting purposes and are available only after veterinary prescription on strict indications. Generally, antimicrobial treatment of animals on individual or on herd basis should not be considered unless in connection with relevant diagnostics. The amounts of antibiotics used and the development of resistance in important pathogens should be closely monitored. Furthermore, resistance monitoring in certain non-pathogenic intestinal bacteria, which may serve as a reservoir for resistance genes is probably more important than hitherto anticipated. Once the usage of or resistance to a certain antibiotic seems to increase in an alarming way, steps should be taken to limit the usage of the drug in order to prevent further spread of resistance genes in animals, humans and the environment. Better methods for detecting and quantifying antibiotic resistance have to be developed. Screening methods must be standardized and evaluated in order to obtain comparable and reliable results from different countries. The genetic mechanisms for development of resistance and spread of resistance genes should be studied in detail. Research in these areas will lead to new ideas on how to inhibit the resistance mechanisms. So far, it has been well established that a heavy antimicrobial drug selective pressure in overcrowded populations of production animals creates favourable environments both for the emergence and the spread of antibiotic resistance genes.

Animal Husbandry↗