Canine rickettsial infections.

Dogs that live in tick-infested areas are at risk for contracting rickettsial infections. Clinical signs associated with ehrlichiosis or Rocky Mountain spotted fever may be dramatic or mild. Clinicians must consider the possibility of rickettsial diseases to request laboratory tests that will permit a proper diagnosis. Specific antimicrobial therapy usually brings about clinical improvement, although some dogs may not be cleared of rickettsial organisms, even with prolonged treatment. A small percentage of dogs die of rickettsial infections, either in the acute stage or owing to chronic bone marrow suppression and generalized debilitation. Ocular lesions are an important clinical sign in canine rickettsial infections and may aid the clinician in making a diagnosis and monitoring response to therapy.     

Diagnostic sampling and establishing a minimum database in exotic animal toxicology.

A toxicologic diagnosis is based on knowledge of the circumstances surrounding a particular case, knowledge of the clinical symptomatology, receipt and evaluation of proper specimens by a qualified laboratory, and judicious interpretation of the laboratory results. Failure to have all necessary ingredients can result in a wrong or missed diagnosis. Many veterinary toxicology laboratories can detect suspected toxicants in feed, tissue, and environmental samples at extremely low concentrations. The ability to detect toxicants at such low levels has often outpaced the ability of the diagnostician to interpret the analytic findings. This article provides guidelines for acquiring a good history, collecting appropriate samples for analysis, and selecting a veterinary analytic laboratory to maximize the probability of making a correct toxicologic diagnosis.     

New perspectives in avian influenza diagnosis

Recent outbreaks of avian influenza (AI) occurring in Europe, in the Americas, in Asia and in Africa have provided field evidence of how challenging the control of this infection can be, particularly in densely populated poultry areas or in areas where free-range rural village poultry and backyard flocks are present. In these areas, laboratory testing is mainly applied to trace viral circulation in a given area or in a susceptible population to implement an early warning system, rather than to diagnose the presence of the virus in a diseased flock or animal. This implies the use of rapid, sensitive and possibly cost-effective laboratory tests adaptable to very high throughputs. As a consequence, new diagnostic approaches and technologies have been increasingly developed and applied. Molecular biology and biotechnology are providing important and precious contributions to the field of AI diagnosis, making extremely rapid, specific and sensitive techniques available. However, the use of some of these technologies is still limited, due to their costs and to the requirement of advanced technical and scientific expertise. Therefore, more conventional and well-established techniques, should not be abandoned but rather reconsidered and improved or modified.     

Real-time polymerase chain reaction: a novel molecular diagnostic tool for equine infectious diseases

The focus of rapid diagnosis of infectious disease of horses in the last decade has shifted from the conventional laboratory techniques of antigen detection, microscopy, and culture to molecular diagnosis of infectious agents. Equine practitioners must be able to interpret the use, limitations, and results of molecular diagnostic techniques, as they are increasingly integrated into routine microbiology laboratory protocols. Polymerase chain reaction (PCR) is the best-known and most successfully implemented diagnostic molecular technology to date. It can detect slow-growing, difficult-to-cultivate, or uncultivatable microorganisms and can be used in situations in which clinical microbiology diagnostic procedures are inadequate, time-consuming, difficult, expensive, or hazardous to laboratory staff. Inherent technical limitations of PCR are present, but they are reduced in laboratories that use standardized protocols, conduct rigid validation protocols, and adhere to appropriate quality-control procedures. Improvements in PCR, especially probe-based real-time PCR, have broadened its diagnostic capabilities in clinical infectious diseases to complement and even surpass traditional methods in some situations. Furthermore, real-time PCR is capable of quantitation, allowing discrimination of clinically relevant infections characterized by pathogen replication and high pathogen loads from chronic latent infections. Automation of all components of PCR is now possible, which will decrease the risk of generating false-positive results due to contamination. The novel real-time PCR strategy and clinical applications in equine infectious diseases will be the subject of this review.     

Diagnostic parasitology

Diagnosis of parasitic infections in small animals is challenging, interesting, and fun, and involves the recognition of parasite stages based on size, morphology, color, and movement. Size and morphology are the major diagnostic parameters, and a calibrated microscope is the essential tool in the diagnostic laboratory. The fecal flotation technique is used routinely for the diagnosis of most parasites that reside within the gastrointestinal tract. The specific gravity and type of solution used often will influence the results obtained. A sugar flotation solution with a specific gravity of 1.27 is one that will float a majority of parasite stages with minimal distortion. Other techniques for diagnosis of specific gastrointestinal parasites include the direct smear and the merthiolate-iodine-formalin preservative method for diagnosis of Giardia sp, the Baermann technique for diagnosis of lungworm and other live larvae, the direct sedimentation technique for diagnosis of trematode eggs, the ether-formalin-sedimentation technique for diagnosis of trematode eggs and concentrating protozoan cysts from feces with high fat content, and the McMaster technique, a dilution modification of the fecal flotation technique. Techniques for evaluation of parasites in blood include the Knott's test, hematocrit method, and direct blood smear for diagnosis of microfilariae and thick and thin stained blood smears for diagnosis of Babesia sp, Haemobartonella sp, Cytauxzoon felis, Hepatozoon canis, and Trypanosoma cruzi. Tissue impression smears are used commonly for the diagnosis of Leishmania sp in dogs and biopsy specimens or aspirates are used for the diagnosis of Pneumocystis carinii. Diagnosis of acute toxoplasmosis can be accomplished with peritoneal and thoracic fluids, and organisms occasionally are detected in blood or spinal fluid. Serological tests for many parasitic diseases have been developed and often are used as supportive diagnostic tests.     

Decision making on helminths in cattle: diagnostics,economics and human behaviour

Helminth infections of cattle affect productivity in all classes of stock, and are amongst the most important production-limiting diseases of grazing ruminants. Over the last decade, there has been a shift in focus in the diagnosis of these infections from merely detecting presence/absence of infection towards detecting its impact on production. This has been facilitated by studies observing consistent negative correlations between helminth diagnostic test results and measures of productivity. Veterinarians are increasingly challenged to consider the economic aspects of their work, and the use of these tests should now be integrated in economic evaluation frameworks for improved decision making. In this paper, we review recent insights in the farm-specific economic impact of helminth infections on dairy cattle farms as well as in farmer attitudes and behaviour regarding helminth control. Combining better economic impact assessments of helminth infections together with a deeper understanding of the non-economic factors that drive a farmer’s animal health decisions should result in more effective control strategies and increased farmer satisfaction.     

Immunologic methods for the detection of humoral and cellular immunity.

A variety of immunologic techniques have been introduced during the past few years. Many of these techniques are being applied to clinical specimens in an attempt to help the practicing veterinarian make a diagnosis. The introduction of new techniques requires extensive testing with clinically normal and diseased patients. It is essential for the practicing veterinarian to understand that the techniques available for the detection of immunologic disorders in the dog and cat are not routine diagnostic procedures and that adequate information has not been developed for any of the techniques described to assure the clinical significance of either positive or negative results (Table 3). This should not discourage the practitioner from submitting samples, but should encourage him or her to question the significance of those results and to attempt to correlate them with history and clinical signs before arriving at a final diagnosis.     

Current knowledge of water quality and safety for livestock.

Basic laboratory evaluation of water quality for livestock should include measurement of TDS, sulfate, nitrate-nitrite, and coliform bacteria. Supplementary water tests may include pH, sodium, iron, magnesium, chloride, calcium, potassium, manganese, and contaminants specific to the situation. Using the best-quality drinking water available contributes to the optimal production of livestock. Restricted quantity of drinking water or drinking water containing excessive levels of nitrate, TDS, sulfate, and other constituents can affect growth and production of all classes of animals. Drinking-water quality and availability should be evaluated as a cause of poor performance or nonspecific disease conditions in livestock. It is important that attempts to evaluate water quality include obtaining a thorough history, making astute observations, and asking intelligent questions. A thorough laboratory examination of animal specimens and water samples should be evaluated in view of existing standards for livestock drinking-water quality.     

Granulomatous dermatitis caused by Mycobacterium genavense in two psittacine birds

Two cases of cutaneous mycobacteriosis in psittacine birds showing featherless, non-painful, non-pruritic nodules are described. Histopathological studies of skin biopsies from both cases demonstrated the presence of a diffuse granulomatous dermatitis with acid- fast organism s inside macrophages, which led to the diagnosis of cutaneous mycobacteriosis. In one case, generalization of the process to internal organs (intestinal and hepatic serosae) was observed. Mycobacterial organisms could not be cultured using conventional isolation media (Coletsos and Löwenstein–Jensen), but polymerase chain reaction (PCR) technique performed on pathological samples from both birds revealed the presence of Mycobacterium genavense . It is thus proposed that cutaneous mycobacteriosis infections, in particular those caused by M . genavense , should be included in the differential diagnosis of skin nodular processes in psittacine birds. The usefulness of PCR techniques for aetiological diagnosis of mycobacterial infections is emphasized.     

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

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

Laboratory procedures for the diagnosis of gastrointestinal tract diseases of dogs and cats

An increasing number of laboratory tests are available for diagnosis of gastrointestinal tract diseases in dogs and cats. Use of these tests can lead to more accurate and rapid diagnoses. This review discusses laboratory tests, both new and old, and the role they currently play in the evaluation of animals presented with gastrointestinal problems. A minimum database helps assess the severity of the disorder, detect extra-gastrointestinal causes of problems and assists in formulating diagnostic and therapeutic plans. Faecal examination remains one of the most important diagnostic procedures in the investigation of gastrointestinal problems. Zinc sulphate faecal flotation is an excellent routine screening technique for helminth and protozoal infections, including giardiasis. Rectal cytology can assist in the diagnosis of large bowel disorders. Interpretation of faecal immunodiagnostic tests is hampered by insufficient knowledge of test sensitivities and specificities. Routine faecal cultures are not warranted and faecal occult blood tests are rarely indicated. Serum tests for gastric inflammation are now under development. The serum trypsin-like immunoreactivity test remains the gold standard technique for the diagnosis of exocrine pancreatic insufficiency. Breath hydrogen tests can be helpful in assessing the functional relevance of mild abnormalities in small-bowel biopsy specimens. Subnormal concentrations of serum cobalamin appear to be more specific indicators of gastrointestinal disease in cats than in dogs. Tests for small intestinal bacterial overgrowth remain controversial and assessment of gastrointestinal permeability has yet to prove its value in the diagnostic assessment of companion animals with gastrointestinal problems. Faecal alpha1-protease inhibitor shows promise for the diagnosis of protein-losing enteropathy.     

Bacterial-associated diarrhea in the dog: a critical appraisal.

The clinical documentation of enteropathogenic bacteria causing diarrhea in dogs is clouded by the presence of many of these organisms existing as normal constituents of the indigenous intestinal flora. The diagnosis of a putative bacterial enteropathogen(s) in dogs should be made based on a combination of parameters, including signalment and predisposing factors, clinical signs, serologic assays for toxins, fecal culture, and PCR. Relying on results of fecal culture alone is problematic, because C perfringens, C difficile, Campylobacter spp, and pathogenic and non-pathogenic E coli are commonly isolated from apparently healthy dogs [10,13,33]. Nevertheless, culture may be useful in procuring isolates for the application of molecular techniques, such as PCR, for detection of specific toxin genes or molecular typing of isolated strains to establish clonality in suspected outbreaks. The oversimplistic attempt to characterize bacterially associated diarrhea by anatomic localization of clinical signs should be discouraged, because most of the previously mentioned bacteria have been associated with small and large intestinal diarrhea. Accurate diagnosis of infections may require diagnostic laboratories to incorporate PCR-based assays using genus- and species-specific primers to facilitate detection of toxin genes and differentiation of species that appear phenotypically and biochemically similar. There has been tremendous interest in the application of microarray technology for the simultaneous detection of thousands of genes or target DNA sequences on one glass slide. This powerful tool could be used for detection of specific pathogenic bacterial strains in fecal specimens obtained from dogs in the future.     

Diagnosis of feline viral infection

A diagnosis of a specific viral disease in the cat involves a combination of an accurate history, careful observation of disease signs, demonstration of characteristic clinical pathologic changes, and isolation or identification of the virus. Isolation or identification of a virus from the patient does not establish that the disease observed was caused by the virus so isolated or identified; correlation and proper interpretation of all findings are necessary to establish a diagnosis. Virus identification may involve office laboratory tests, such as cytology or ELISA, or more specialized procedures. Whether specimens are to be sent out for specialized tests or office laboratory procedures are to be used, the veterinary practitioner must not only know what specimens are required but must also understand the test and be able to properly interpret the results in light of the patient's observed condition.     

Malassezia dermatitis and otitis.

The incidence of dermatitis and otitis resulting from overgrowth of M. pachydermatis is great enough that cytological sampling techniques should be considered a routine part of the dermatological examination. Because most cases of MD and Malassezia otitis cannot be grossly distinguished from bacterial pyoderma and otitis, respectively, efficiency in performing cytology testing of skin and ear canal exudate is essential to the successful diagnosis and management of pruritic skin diseases and otitis. Although Malassezia infections are rarely primary, therapy can be instituted to remove the yeast as a confounding factor while a differential diagnosis is pursued in evaluating the underlying disease process.     

Abdominal, thoracic, and pericardial effusions.

The laboratory evaluation of abdominal, thoracic, and pericardial effusions is a useful diagnostic tool for the assessment of disease states that result in fluid accumulation. Although the numeric values pertaining to cell count and protein content are important, the microscopic evaluation is a critical aspect of the diagnostic procedure; not only does it allow complete classification of the fluid but it allows identification of specific cell types or microorganisms that might be responsible for the fluid accumulation. These findings should always be interpreted in conjunction with the history, signalment, physical findings, and other diagnostic aids in making a definitive diagnosis.     

Epidemiology of animal diseases in Africa: approach strategies and role of veterinary laboratories

Study of the epidemiology of animal diseases in Africa should only be envisaged when the veterinary diagnosis laboratories and the cattle development structures in the field are fully operational. Veterinary epidemiology may be approached using all usual laboratory techniques. In spite of some limitations described by the author, current diagnosis, intervention in outbreaks, retrospective and prospective surveys all contribute to the knowledge of animal disease epidemiology. In fact, considering the difficulties met in Africa, it seems advisable to combine a mixed approach to maximise the use of field visits. The author concludes that until new techniques allow a decentralised diagnosis, the African veterinary laboratories must play a determinant part in the study of the epidemiology of animal diseases; furthermore current diagnosis, sanitary surveillance and epidemiological surveys have now become almost inseparable.     

Fungal diseases of amphibians: an overview.

Clinicians should be familiar with the most common fungal diseases of amphibians. Because lesions in mycotic diseases are nonspecific, a diagnosis cannot be established solely on the basis of clinical presentation. Bacterial, mycobacterial, chlamydial, and parasitic infections, and toxic or environmental conditions may mimic mycotic disease to various extents. Furthermore, mycoses may be masked by overwhelming secondary bacterial infection and therefore remain undiagnosed. Skin scrapings, impression smears, biopsies, and fungal culture are all useful tools in confirming or dismissing a diagnosis of mycosis. Whenever possible, an effort should be made to forward samples and biopsies for culture to appropriate laboratories. Providing the laboratory with a tentative etiologic diagnosis may allow for specific selection of more specific agars and culture conditions and maximize the chances of recovering the fungus from lesions. Identification to species level should also be encouraged, if progress is to be made in the understanding of mycoses in amphibians. The morphology of an isolate should be consistent with the microscopic features of the fungus in histological sections of affected tissues, if it is to be firmly incriminated as the cause of disease. A complete necropsy should be conducted on animals that die or are found dead, and, ideally, isolates from confirmed cases of fungal infection should be deposited in scientific collections, so that they are available for later studies. In addendum, readers should be aware that there is recent evidence to suggest that at least some published cases of amphibian basidiobolomycosis were in fact cases of chytridiomycosis [38], and therefore the validity of basidiobolomycosis as a disease entity in amphibians may be revisited in the years to come.     

Laboratory procedures for the diagnosis of gastrointestinal tract diseases of dogs and cats

An increasing number of laboratory tests are available for diagnosis of gastrointestinal tract diseases in dogs and cats. Use of these tests can lead to more accurate and rapid diagnoses. This review discusses laboratory tests, both new and old, and the role they currently play in the evaluation of animals presented with gastrointestinal problems. A minimum database helps assess the severity of the disorder, detect extra-gastrointestinal causes of problems and assists in formulating diagnostic and therapeutic plans.

Faecal examination remains one of the most important diagnostic procedures in the investigation of gastrointestinal problems. Zinc sulphate faecal flotation is an excellent routine screening technique for helminth and protozoal infections, including giardiasis. Rectal cytology can assist in the diagnosis of large bowel disorders. Interpretation of faecal immunodiagnostic tests is hampered by insufficient knowledge of test sensitivities and specificities. Routine faecal cultures are not warranted and faecal occult blood tests are rarely indicated.

Serum tests for gastric inflammation are now under development. The serum trypsin-like immunoreactivity test remains the gold standard technique for the diagnosis of exocrine pancreatic insufficiency. Breath hydrogen tests can be helpful in assessing the functional relevance of mild abnormalities in small-bowel biopsy specimens. Subnormal concentrations of serum cobalamin appear to be more specific indicators of gastrointestinal disease in cats than in dogs. Tests for small intestinal bacterial overgrowth remain controversial and assessment of gastrointestinal permeability has yet to prove its value in the diagnostic assessment of companion animals with gastrointestinal problems. Faecal alpha₁-protease inhibitor (α₁-PI) shows promise for the diagnosis of protein-losing enteropathy.