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Fimbrial adhesions of Escherichia coli.

Fimbriae are long, threadlike protein polymers found on the surface of many strains of Escherichia coli. The presence of fimbriae has been found to be significantly correlated with pathogenicity, and specific fimbriae confer on pathogenic strains the ability to adhere to and colonize various specific host epithelia. A surprising variety of fimbrial adhesions have been found in E. coli; several unrelated groups of such proteins have been extensively characterized. Fimbriae of E. coli are strongly immunogenic. Their continuous interaction with host defenses has imposed heavy selective pressure on them, leading to a fine balance between functional necessity and maximal immunologic diversity. These important virulence factors may be exploited in various ways in the production of vaccination agents. Vaccines based on E. coli fimbriae have proved extremely successful in the veterinary sector, and several types of immunization are currently being tested in humans.

Adhesins, Escherichia coli↗

Rapid detection of group C streptococci from animals by latex agglutination.

A group C latex agglutination reagent, included as the negative control in the PathoDx Strep A latex agglutination test (Diagnostic Products Corp., Los Angeles, Calif), was compared with culture for rapid detection of group C streptococci (Streptococcus equi, S. equisimilis, S. zooepidemicus, and S. dysgalactiae) from swabs of veterinary specimens. The overall sensitivity of the latex test was 78%, and specificity was 97.6%. Only 25% of S. dysgalactiae isolates were detected, thereby accounting for the relatively low sensitivity. Ninety-three percent of the group C streptococci other than S. dysgalactiae were isolated from horses. When the latex test was evaluated for detection of group C streptococci other than S. dysgalactiae from horses, the sensitivity and specificity were 95.3 and 100%, respectively. We found the group C latex agglutination test to be a rapid and accurate method for the detection of the major pathogenic group C streptococci from swabs of equine specimens.

Animals↗

The epidemiology of avian herpesviruses in veterinary medicine.

There are ten avian herpesviruses, which have been isolated from eight orders. Six of these are of veterinary importance: Pacheco's parrot disease virus, pigeon herpesvirus, duck plague virus, infectious laryngotracheitis virus, herpesvirus of turkeys and Marek's disease virus. The knowledge on the epidemiology of each virus and the disease it causes is discussed. Features in common to infections with most avian herpesviruses are: infection is persistent in individuals and ubiquitous in populations; virus is shed for long periods of time after infection although in some cases erratically; infection does not necessarily result in disease and at least in some avian herpesvirus infections the incidence of disease is affected by the pathogenicity of the virus; the genetic constitution of the host and stress factors affecting the host. It is concluded that man's interference with the natural history of host species often increases the threat and incidence of disease unless preventive action is taken.

Animals↗

Accuracy of polymerase chain reaction assays for diagnosis of feline immunodeficiency virus infection in cats.

OBJECTIVE: To determine the sensitivity, specificity, and overall diagnostic accuracy of polymerase chain reaction (PCR) assays offered by commercial diagnostic laboratories for diagnosis of FIV infection in cats. DESIGN: Prospective clinical trial. ANIMALS: 124 cats. PROCEDURE: Blood was collected from cats that were neither infected with nor vaccinated against FIV, uninfected cats that were vaccinated with a licensed FIV vaccine, and cats experimentally and naturally infected with FIV representing subtypes A, B, and C. Coded blood samples were submitted to 3 laboratories in the United States and Canada offering PCR assays for diagnosis of FIV infection to veterinary practitioners. All laboratories tested fresh blood samples, and 1 laboratory also tested samples submitted as dried blood smears. The FIV infection status in all cats was confirmed by virus isolation. Sensitivity, specificity, and correct results were calculated for each PCR assay. RESULTS: Sensitivity ranged from 41% to 93%. Specificity ranged from 81% to 100% in unvaccinated cats and 44% to 95% in cats vaccinated against FIV. Correct results were obtained in 58% to 90% of 124 cats tested. All tests misidentified both uninfected and infected cats. False-positive results by all laboratories were higher in cats vaccinated against FIV than in unvaccinated cats, suggesting that vaccination interferes with the performance or interpretation of PCR assays used for diagnosis of FIV infection. CONCLUSIONS AND CLINICAL RELEVANCE: PCR assays used for diagnosis of FIV infection presently marketed to veterinary practitioners in North America vary significantly in diagnostic accuracy and did not resolve the diagnostic dilemma resulting from vaccination of cats against FIV.

Animals↗

Cryptosporidiosis in zoo and pet birds.

Cryptosporidiosis is recognized as a primary disease in commercially raised chickens, turkeys, and bobwhite quail. Little is known about cryptosporidial infections in zoo and pet birds, although, infections of the small intestines, proventriculus, respiratory tract, and kidneys have been reported. In the present study, we reviewed cases of cryptosporidial infections in zoo and pet birds submitted to the State Veterinary Diagnostic Laboratory, Auburn, Alabama for necropsy or histopathologic examination. We identified infections in cockatiels, white-lored euphonias, bronze mannikin finches, and Australian diamond firetailed finches. Infections in the cockatiels occurred mostly in the small intestine, but parasites were also observed in the esophageal glands, air sacs, and proventriculus of some birds. Separate cases of small intestinal and proventricular infections were identified in the white-lored euphonias. Cryptosporidial parasites were found only in the proventriculus of the bronze mannikin finches and Australian diamond firetail finches. No cases of renal cryptosporidiosis were observed. Co-pathogens or other disease conditions were present in all birds. The Cryptosporidium species responsible for causing proventricular infection in zoo and pet birds may be different from C. meleagridis and C. baileyi.

Animals↗

Emerging tick-borne viral diseases in East Asia: pathology-driven insights into pathogenesis and disease causality.

SUMMARYTick-borne viral infections have emerged as a significant and growing public health concern. In East Asia, severe fever with thrombocytopenia syndrome (SFTS) has served as a prototypical disease in which pathological analyses have substantially advanced the understanding of disease pathogenesis. SFTS is characterized by profound immune dysregulation driven by viral tropism for plasmablast-lineage B cells, leading to cytokine storm and hemophagocytic syndrome. Complementary analyses of human clinical specimens and experimental animal models, including cats and ferrets, have provided critical insights into the immunopathogenesis of SFTS. The recent identification of additional tick-borne viruses, including Oz virus (OZV), Yezo virus (YEZV), and Alongshan virus (ALSV), has further expanded the spectrum of emerging infections in this region. Notably, pathological investigation of a fatal human case of OZV infection demonstrated direct viral localization within cardiomyocytes, establishing a causal link between infection and fulminant myocarditis and highlighting a distinct organ-specific pathogenic mechanism. Despite the advances in genomic technologies that enable rapid detection of novel viruses, establishing causal relationships between viral presence and disease remains a major challenge. Tissue-based pathological approaches, particularly in situ localization of viral components, are, therefore, essential for defining disease mechanisms and confirming etiological roles. This review provides a comprehensive synthesis of tick-borne viral infections in East Asia, with particular emphasis on Japan, integrating pathological, virological, and clinical perspectives. It also identifies key knowledge gaps and underscores the importance of a synergistic One Health framework that incorporates both human and veterinary pathology to advance the understanding and control of these emerging diseases.

One Health↗

An outbreak of contagious bovine pleuropneumonia in the Owambo Mangetti area of South West Africa/Namibia: microbiological, immunofluorescent, pathological and serological findings.

An outbreak of contagious bovine pleuropneumonia (CBPP) in the Owambo Mangetti area of South West Africa/Namibia in 1982 led to the slaughter of 3,153 cattle. The lungs of 266 of these animals were scrutinized for gross lesions of CBPP and samples of lung tissue, sequestral contents or pleural fluid were submitted to the Veterinary Research Institute (VRI), Onderstepoort for microbiological, immunofluorescent and pathological examination. Immunofluorescence proved to be the most successful method of diagnosis producing 96% of positives, while mycoplasma isolations were only positive in 64% of the 55 specimens processed in parallel. This clearly demonstrated the value of the former technique in the accurate as well as rapid diagnosis of CBPP. The impression smear technique employed, using Eriochrome black counterstaining proved most satisfactory and easy to interpret. The isolation of M. mycoides was influenced by the transit times and temperature of the samples on arrival at the VRI. No pathogenic bacteria were found in routine aerobic bacterial cultures from 27 of the samples submitted. Complement fixing antibodies were present in the sera of 16 cattle and titres varied between 10 and 320. Animals in which antibodies were absent included those with early lung lesions and some with sequestra.

Animals↗

Antimicrobial agents are societal drugs: how should this influence prescribing?

This paper is concerned with how those who prescribe antimicrobials should consider the wider repercussions of their actions. It is accepted that in an ecological system, pressure will cause evolution; this is also the case with antimicrobials, the result being the development of resistance and the therapeutic failure of drugs. To an extent, this can be ameliorated through advances by the pharmaceutical industry, but that should not stop us from critically appraising our use and modifying our behavior to slow this process down. Up to 50% of prescribing in human medicine and 80% in veterinary medicine and farming has been considered questionable. The Alliance for the Prudent Use of Antimicrobials (APUA) was approached by the WHO to review the situation. Their recommendations include decreasing the prescribing of antibacterials for nonbacterial infections. In the UK, there has been an initiative called "the path of least resistance". This encourages general practitioners to avoid prescribing or reduce the duration of prescriptions for conditions such as upper respiratory tract infections and uncomplicated urinary tract infections; this approach has been successful. Another recommendation is to reduce the prescribing of broad-spectrum antibacterials. In UK hospitals, the problems identified with the inappropriate use of antibacterials are insufficient training in infectious disease, difficulty in selecting empirical antibacterial therapy, poor use of available microbiological information, the fear of litigation and the fact that the majority of antibacterials are prescribed by the least experienced doctors. With close liaison between the laboratories and clinicians, and the development of local protocols, this can be addressed. Another recommendation is to tighten the use of antibacterial prophylaxis and to improve patient compliance. Through a combination of improved education for doctors and patients, and improved communication skills, these problems can be addressed. A further recommendation is to encourage teaching methods that modify prescribing habits. It has been shown that workshops have led to a significant reduction in the prescribing of broad-spectrum antibacterials in the community. Auditing the prescribing of antibacterials has also been recommended. Surveillance systems around the world monitor trends in resistance: the European Antimicrobial Resistance Surveillance Progamme (EARSS) monitors antibacterial resistance; the WHO and the International Union Against Tuberculosis and Lung Disease collaborate to monitor tuberculosis; the WHO and the International AIDS Society monitor HIV. In the third world, a bigger problem than resistance is whether drugs are even effective, as they are often spoiled by climactic conditions, and poor quality generics and counterfeit drugs are common. Also, patients may not be able to complete a course for financial reasons. Facts about Antimicrobial Resistance in Animals (and Agriculture) and Impact on Resistance (FAAIR) was commissioned by APUA. They conclude that the nonhuman use of antibacterials can lead to the development of antibacterial resistance in human pathogens. The European commission banned the use of antibacterials as growth promoters in 1999. In the Western world, we should improve our diagnosis of sepsis, access local guidelines and consider withholding treatment pending investigations, decide if treatment can be stopped earlier and treat the patient not the result. Many developing countries need improved access to more antimicrobials, preferably in the controlled environment of appropriate medical advice.

Animals↗

Pathogenic intestinal parasites and bacterial agents in solid wastes.

OBJECTIVE: To determine the profile of potentially pathogenic enteric parasites and bacterial agents in municipal refuse dumps in Ibadan, Nigeria. DESIGN: A cross-sectional survey. SETTING: Five major market places refuse dumps in Ibadan municipality, Nigeria. METHODS: The major market places in Ibadan city were randomly selected by lottery method. The refuse sludge were sampled and examined parasitologically and bacteriologically using the methods as described. Data analysis was done by using chi-square test where applicable. RESULTS: Cases of multiple intestinal parasites and bacterial agents were commonly encountered in the sludge refuse samples. The commonly found parasitic agents were of both human and veterinary importance. These include Ascaris Lumbricoides (9.3 egp), Entamoeba histolytica (8.07 cyst per gram), Hookworm/strongyle (6.27 egp) and Ascaris suum (1.07 egp). Others were Ascaris vitolorum (1.09 epg), Strongyloides papillosu (0.52 larvae per gram.), Schistosoma suis (0.31 epg) Dicrocoelium dendriticum (0.9 egp). The most commonly found bacterial agents were Klebsiella species, Escherichia Coli, Proteus species, streptococci, and other gram-positive organisms. Climatic conditions affected the distribution of both parasites and bacterial agents in the sludge (P<0.001). More intestinal parasites 2423 (53.4%) and bacterial agents 2150 (27.2%) were encountered at mean air temperature 26.1 +/- 0.6, mean relative humidity of 72 +/- 3.5%. The degree of contamination by market locations varies significantly (P<0.001). CONCLUSION: A high degree of contamination of solid waste dumpsites with bacterial and parasitic agents was observed in the present study. As a result of the public health importance of the organisms isolated it is opined that well planned waste management and health education programs will go a long way to reduce the potential epidemic risks posed by such sites in Ibadan, Nigeria.

Animals↗

Murine model of pregnancy-associated Listeria monocytogenes infection.

Listeria monocytogenes has been recognized as a significant pathogen, occurring worldwide, capable of causing animal and human infections. In its most severe form, listeriosis is an invasive disease that affects immunocompromised patients. Additionally, pregnant women represent a high-risk group for L. monocytogenes infection. Abortion, stillbirth or severe neonatal infection can be the serious outcome of such an infection. In an experimental murine model of pregnancy-associated listeriosis we studied the impact of L. monocytogenes on the maternal immune response and pregnancy outcome. In comparison to virgin animals, pregnant mice mounted lower levels of protective cytokines and were unable to eliminate the pathogen. The impaired maternal immune response that has been found both on the systemic and local level, facilitated bacterial multiplication in the liver, placenta and ultimately in the fetal tissues. This resulted in severe necrotizing hemorrhagic hepatitis and Listeria-induced placental necrosis, increasing the incidence of postimplantation loss and poor pregnancy outcome.

Abortion, Veterinary↗

[Udder health on dairy farms. 2. Mastitis prevention programs].

Herd-specific mastitis control programmes are based on the continuous monitoring of primary parameters, of which the bulk milk somatic cell count and the distribution of pathogens responsible for clinical and subclinical mastitis are the most important ones. When a mastitis control programme is started, the herd situation should be analysed. Sampling milk from cows with clinical and subclinical mastitis is essential. During the last 5 years, a number of programmes related to udder health and milk quality have been started in the Netherlands. These programmes are: 1) 'Chain Quality Milk', 2) a herd-specific Mastitis Planner, 3) a sampling programme for subclinical mastitis, and 4) a combined analysis of milk recording data, and subclinical and clinical mastitis data. Using these four programmes, farmers, veterinary practitioners, and other advisors can design a herd-specific programme to improve udder health. Although there have been many studies on mastitis and important progress has been made, some important aspects have often not been studied. Suggestions for further research are made.

Animals↗

Characterization of a major outer membrane protein multigene family in Ehrlichia ruminantium.

Ehrlichia ruminantium is a tick-transmitted rickettsial pathogen, which causes heartwater or cowdriosis in wild and domestic ruminants. A dominant antibody response of animals infected with E. ruminantium is directed against the outer membrane protein MAP1 (major antigenic protein 1). Part of the locus containing map1 has been characterized and consists of four map1 paralogs, designated map1-2, map1-1, map1 and map1+1, indicating that map1 is encoded by a multigene family. The purpose of this study was to determine the total number of map1 paralogs and their transcriptional activities. Using genome walking and data from an ongoing E. ruminantium genome sequencing project at the Onderstepoort Veterinary Institute, we found 16 paralogs of the map1 gene tandemly arranged in a 25 kb region of the E. ruminantium genome. The map1 multigene family is downstream of a hypothetical transcriptional regulator gene and upstream of the secA gene. Thirteen paralogs at the 5' end of the 25-kb locus were connected by short intergenic spaces (ranging from 0 to 42 bp) and the remaining three paralogs at the 3' end were connected by longer intergenic spaces (ranging from 375 to 1612 bp). All 16 map1 paralogs were transcriptionally active in E. ruminantium grown in endothelial cells and paralogs with short intergenic spaces were co-transcribed with their adjacent genes.

Antigens, Bacterial↗

Detection of tetracycline resistance genes by PCR methods.

Rapid, accurate, and sensitive determination of antibiotic resistance profiles of various human and animal pathogens becomes a vital prerequisite for successful therapeutic intervention in the face of the increased occurrences of drug-resistant bacterial infections. The current methods, which are dependent on cultivation of pathogens and phenotypic expression of antibiotic resistance, usually require excessive time, special microbiological equipment, and qualified personnel. However, even with all these requisites, for example, no bacteria can be grown from more than 80% of all clinical samples sent to clinical microbiology laboratories. Besides the cultivation limitations, the cultivation-based determination of an antibiotic resistance profile lacks the genotypic information, which is essential for understanding the epidemiology and routes of transmission of antibiotic resistance genes. These genes often reside on mobile genetic elements and can move freely between commensal and pathogenic microbiota, occurring even between taxonomically distant clinical and environmental microbiota. Therefore, development of genotyping methods for detection of antibiotic resistance genes is highly desirable for fast, accurate, and sensitive detection of antibiotic resistance genes in a broad range of pathogenic and commensal bacteria in both clinical and environmental samples. As a model for our studies we have chosen the genes conferring resistance to tetracyclines. Tetracyclines belong to a family of broad-spectrum antibiotics that include tetracycline, chlortetracycline, oxytetracycline, demeclocycline, methacycline, doxycycline, minocycline, and a number of other semisynthetic derivatives. These antibiotics inhibit protein synthesis in Gram-positive and Gram-negative bacteria by preventing the binding of aminoacyl-tRNA molecules to the 30S ribosomal subunit. The antibiotics of this group were introduced in the late 1950s and since then have been widely used in clinical and veterinary medicine, as well as for prophylaxis and growth promotion in food animals. Because of the possible misuse and overuse of these drugs, resistance to this class of antibiotics is widespread among many clinical isolates, thus limiting the utility of tetracyclines in treating infections. Despite this shortcoming, antibiotics of this class still remain in the active arsenal for dermatologists to treat skin infections such as acne and rosacea.

Animals↗

Oral vaccination of wildlife against rabies: opportunities and challenges in prevention and control.

Rabies is an acute, progressive, fatal encephalitis caused by viruses in the Family Rhabdoviridae, Genus Lyssavirus. Rabies virus is the representative member of the group. Warm-blooded vertebrates are susceptible to experimental infection, but major primary hosts for disease perpetuation encompass bats and mammalian carnivores. The dog is the global reservoir, and important wild carnivores include foxes, raccoons, skunks, and mongoose, among others. Traditionally, reliance upon long-term, widespread, government-supported programmes aimed at population reduction of animals at risk has been unsuccessful as the sole means of rabies control, based in part upon economical, ecological and ethical grounds. In contrast, immunization of domestic dogs with traditional veterinary vaccines by the parenteral route led to the virtual extinction of canine-transmitted rabies in developed countries. Taken from this basic concept of applied herd immunity, the idea of wildlife vaccination was conceived during the 1960s, and modified-live rabies viruses were used for the experimental oral vaccination of carnivores by the 1970s. The development of safe and effective rabies virus vaccines applied in attractive baits resulted in the first field trials in Switzerland in 1978. Thereafter, technical improvements occurred in vaccine quality and production, including the design of recombinant viruses, as well as in the ease of mass distribution of millions of edible baits over large geographical areas. Over the past few decades, extensive oral vaccination programmes focusing upon the red fox, using hand and aerial distribution of vaccine-laden baits, have resulted in the virtual disappearance of rabies in Western Europe. The same dramatic observation held true for southern Ontario. During the 1990s in the United States, oral vaccination programmes concentrated upon raccoons, grey foxes, and coyotes, with similar success. For example, raccoon rabies has not spread west of the current focus in the eastern states, grey fox rabies is contained in west central Texas, and no recent cases of rabies have been reported from coyotes away from the Mexican border for several years. Despite the progress observed and the absence of substantive adverse environmental or health effects, oral vaccination is not a panacea, and should be viewed as an important adjunct to traditional prevention and control techniques in human and veterinary medicine. Local outbreak suppression of rabies among free-ranging wildlife is documented, and regional elimination of particular virus variants among specific, targeted carnivore hosts is demonstrable, but true disease eradication is not achievable at the present time by current techniques. For example, no practical vaccination methods have been designed for bats. Although lyssaviruses appear in relative compartmentalization between the Chiroptera and Carnivora, major spillover events have been detected from bats to carnivores, and phylogenetic analyses suggest a historical basis for extant viral origins due to interactions between these taxa. Thus, bio-political considerations aside, the possibility for pathogen emergence resulting from transmission by rabid bats with subsequent perpetuation among other animals cannot be discounted easily on any continent, with the possible exception of Antarctica. Clearly, given their biodiversity, distribution, and abundance, novel methods would be necessary to consider meaningful control of rabies in these unique volant mammals. Newer approaches in biotechnology may be envisaged some day for eventual extension to bats, as well as more widespread application to global canine rabies remediation in developing countries.

Administration, Oral↗

Reproductive losses caused by bovine viral diarrhea virus and leptospirosis.

Bovine viral diarrhea virus and Leptospira spp. are two of the common pathogenic organisms responsible for reproductive losses in cattle worldwide. Both can be come endemic in herds resulting in chronic low-grade reproductive losses or they can be introduced into relatively naïve herds, resulting in substantial reproductive losses over a short period of time. Both organisms are a differential diagnoses for common reproductive losses that veterinarians investigate, including low conception rates and abortions.

Abortion, Veterinary↗

Preliminary analysis of tetracycline residues in marketed pork in Hanoi, Vietnam.

A cross-sectional survey was designed to investigate the proportion of tetracycline residues in marketed pork in suburb and urban districts in Hanoi. A total of 290 raw muscle samples were randomly collected from open markets in these districts. The samples were qualitatively screened for tetracycline residues using the agar inhibition test, and Bacillus cereus (ATCC 11778) as the reference strain. The inconclusive samples were then analyzed using high-performance liquid chromatography (HPLC). The positive samples from either test were defined as positive results. Overall, 5.5% of all collected samples were positive for tetracycline residues. The proportion of positive samples from shops in suburb districts was significantly (P < 0.05) different from those collected from shops in urban districts. So, the factor of region was identified as a risk factor of tetracycline residue proportion in raw pork with an odds ratio (OR) of 4.03 (95% CI = 1.12, 14.45). For the other factors, such as season, type of shop, type of abattoir, origin of meat, etc., the difference in proportion of positive samples within each factor was substantial but not statistically significant. These factors were identified as nonrisk factors. Such a high proportion may pose a potential hazard to public health, particularly since they might induce drug resistance of pathogenic micro-organisms.

Abattoirs↗

[Coxiella burnetii: what is the reality?].

After the Second World War, in Italy Q Fever or Coxiellosis has been shown a significant relevance, a recrudescence with an epidemic state for over ten years. Later, the infectious disease occurred as endemic since the 80s, the outbreaks were just isolated. Workflows analysis of some authors has demonstrated the spread out of the infection throughout Italian herds with a prevalence ranging from 1.2 per cent to 10 per cent. Our survey carried out throughout Campania area in cattle has shown a real positivity over 14 per cent performing the IFAT for the detection of IgG antibodies for Coxiella burnetii. Therefore, it has been so important to stress the influence of cattle farming management in stables as a real risk of Coxiellosis. For example, the Relative Risk (RR) has been registrated about 6.84 (2.18<RR<21.4) in comparison with some herds permanently housed than those kept unhoused and about 8.4 (1.8<RR<38.6) in housed herds and those permanently kept at pasture. Therefore, we have detected a seroprevalence of Q Fever about 11.8 per cent within sheep and about 6.3 per cent within goats. The investigations have focused buffaloes too, tested by several authors, especially from India, but in Italy, only Galiero has recorded significant results: the overall seroprevalence (1.2 per cent) was observed within 1012 buffaloes. Other studies have demonstrated that dogs may transmit Q Fever to humans by infected birth fluids and membranes and by urine. Because of this observation, addressed some of our search projects to assess possible relation among other pathogens, such as Leishmania infantum, Rickettsia conori and Ehrlichia canis. The results have confirmed the presence of C. burnetii (seroprevalence about 7 per cent) within tested dogs in southern Italy. There was no direct estimated relationship between C. burnetii and E. canis and R. conori, instead of significant relation has been shown for L. infantum. The investigation focused on the direct interaction between seropositivity and the age of ruminants. For example, the highest prevalence has been observed in cows and sheep ranging from 3-5 years in which the infection risk was higher than in younger herds. In accordance with the workflows serological analysis, C. burnetii is widespread in Italy among housed ruminants. PCR assay was, therefore performed to test cows with clinical signs of abortion and neonatal mortality. All the survey involved 305 animals (267 foeti and 38 stillborn) with 77 positive herds showing DNA of C. burnetii. To detect C. burnetii pathogen in milk samples, two methods have been combined: the immunomagnetic separation (IMS) and PCR assay, obtaining an increasing of sensitivity (10(-17)) more than only PCR technique (10(-8)). The overall analysis of serological and biomolecular results has demonstrated that C. burnetii is really widespread and may have detrimental effects on farm management system.

Abortion, Veterinary↗

Differentiation of Serpulina species by NADH oxidase gene (nox) sequence comparisons and nox-based polymerase chain reaction tests.

The NADH oxidase genes (nox) of 18 strains of intestinal spirochaetes were partially sequenced over 1246 bases. Strains examined included 17 representatives from six species of the genus Serpulina, and the type strain 513A(T) of the human intestinal spirochaete Brachyspira aalborgi. Sequences were aligned and used to investigate phylogenetic relationships between the organisms. Nox sequence identities between strains within the genus Serpulina were within the range 86.3-100%, whilst the nox gene of B. aalborgi shared between 78.8-83.0% sequence identity with the nox sequences of the various Serpulina strains. A phenogram produced based on sequence dissimilarities was in good agreement with the current classification of species in the genus Serpulina, although an atypical strongly beta-haemolytic porcine strain (P280/1), previously thought to be S. innocens, appeared distinct from other members of this species. Primer pairs were developed from the nox sequence alignments for use in polymerase chain reaction (PCR) identification of the pathogenic species S. hyodysenteriae (NOX1), S. intermedia (NOX2), and S. pilosicoli (NOX3), and for the combined non-pathogenic species S. innocens and S. murdochii (NOX4). The PCRs were optimised using 80 strains representing all currently described species in the genus Serpulina, as well as the type strain of B. aalborgi. Tests NOX1 and NOX4 specifically amplified DNA from all members of their respective target species, whilst tests NOX2 and NOX3 were less sensitive. NOX2 amplified DNA from all 10 strains of S. intermedia from pigs but from only 4 of 10 strains from chickens, whilst NOX3 amplified DNA from only 18 of 21 S. pilosicoli strains, even at low stringency. Tests NOX1 and NOX4 should prove useful in veterinary diagnostic laboratories, whilst NOX2 and NOX3 require further refinement.

Animals↗