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Topic:Diagnostic Technique

Diagnostic techniques in equine medicine encompass a range of procedures and tools used to identify diseases, injuries, or other health conditions in horses. These techniques include imaging methods such as radiography, ultrasonography, and magnetic resonance imaging (MRI), as well as laboratory tests like blood work and tissue biopsies. Each diagnostic method provides specific information that can aid in the assessment and management of equine health issues. Radiography is commonly used for evaluating bone structures, while ultrasonography is useful for soft tissue examination. MRI offers detailed images of both soft and hard tissues, though its use is limited by cost and availability. Laboratory tests can detect biochemical and hematological changes indicative of disease. This page compiles peer-reviewed research studies and scholarly articles that explore the development, application, and efficacy of various diagnostic techniques in equine veterinary practice.
Construction and validation of a risk-screening questionnaire for the investigation of recurrent airway obstruction in epidemiological studies of horse populations in Great Britain.
Preventive veterinary medicine    February 20, 2006   Volume 75, Issue 1-2 8-21 doi: 10.1016/j.prevetmed.2006.01.001
Hotchkiss JW, Reid SW, Christley R.Recurrent airway obstruction (RAO) is an environmental respiratory disease affecting horses. A risk-screening questionnaire (RSQ) for RAO would provide a useful tool to investigate the epidemiology of the disease in horses; our aim in this study was to construct and validate such an instrument. Guidance for what questions to include in the RSQ came from three processes: a review of the scientific literature, a survey of equine practitioners in the UK and a consultation with 19 experts using a modified Delphi technique. The latter consultation consisted of two rounds; agreement amongst the expe...
A 1.3-Mb interval map of equine homologs of HSA2.
Cytogenetic and genome research    February 18, 2006   Volume 112, Issue 3-4 227-234 doi: 10.1159/000089875
Wagner ML, Raudsepp T, Goh G, Agarwala R, Schaffer AA, Dranchak PK, Brinkmeyer-Langford C, Skow LC, Chowdhary BP, Mickelson JR.A comparative approach that utilizes information from more densely mapped or sequenced genomes is a proven and efficient means to increase our knowledge of the structure of the horse genome. Human chromosome 2 (HSA2), the second largest human chromosome, comprising 243 Mb, and containing 1246 known genes, corresponds to all or parts of three equine chromosomes. This report describes the assignment of 140 new markers (78 genes and 62 microsatellites) to the equine radiation hybrid (RH) map, and the anchoring of 24 of these markers to horse chromosomes by FISH. The updated equine RH maps for ECA...
Cloning and expression of 51-kDa antigenic protein of Neorickettsia risticii NR-JA1.
Annals of the New York Academy of Sciences    February 17, 2006   Volume 1063 246-251 doi: 10.1196/annals.1355.038
Park MK, Kim EH, Cho MR, Yi YH, Lee MJ, Shah DH, Park JH, Park BK, Eo SK, Lee JH, Chae JS.Neorickettsia (Ehrlichia) risticii is a causative agent of acute diarrheal syndrome in horses, commonly known as Potomac horse fever. Korean isolate of N. risticii NR-JA1 was cultivated in mouse macrophage cell line P388D1. A complete ORF of p51 antigenic protein gene was amplified and cloned into pQE32 and pcDNA3.1 vectors and the resultant clones were named as pQE32/Nr-51 and pcDNA3.1/Nr-51, respectively. Recombinant p51 (rp51) protein antigen was expressed in E. coli (pQE32/Nr-51) and cos-7 cell line (pcDNA3.1/Nr-51). The rp51 protein showed immunoreactivity with anti- mouse p51 antibodies....
Development of an indirect enzyme-linked immunosorbent assay for detecting equine serum antibodies to the lipopolysaccharide of Salmonella abortusequi.
Research in veterinary science    February 17, 2006   Volume 81, Issue 2 215-217 doi: 10.1016/j.rvsc.2005.11.004
Gall D, Nielsen K, Bermudez RM, Muñoz del Real MC, Halbert G, Groulx R, Moreno F, Chow EY, Checkley SL.An indirect enzyme-linked immunosorbent assay (IELISA) was developed for the detection of equine serum antibodies to lipopolysaccharide of Salmonella enterica subsp. enterica serovar Abortusequi (LPS), a causative organism of Equine Paratyphoid. The data presented demonstrates that horses immunized with S. abortusequi LPS developed antibodies detectable by the IELISA. By comparison, the tube agglutination test (TAT) did not detect antibody to S. abortusequi LPS as consistently as the IELISA. The data suggests that the IELISA may be a more suitable test for the detection of serum antibodies to ...
Detection of equine herpesvirus-specific effector and memory cytotoxic immunity in the equine upper respiratory tract.
Veterinary immunology and immunopathology    February 10, 2006   Volume 111, Issue 1-2 117-125 doi: 10.1016/j.vetimm.2006.01.014
Breathnach CC, Yeargan MR, Timoney JF, Allen GP.Immunological protection of horses from equine herpesvirus-1 (EHV-1) infection and disease depends on the cooperation of virus-specific humoral and cellular immune responses. EHV-specific mucosal immunity may be an important component of such immune responses. This study demonstrates the induction of anti-EHV cytotoxic cellular immune responses in various mucosal and systemic lymphoid tissues associated with the upper respiratory tract (URT) of the horse. Four young horses (1-2 years of age) were inoculated intranasally with the Army 183 strain of EHV-1 and euthanized 1 week later. One untreat...
A monoclonal antibody to equine interleukin-4.
Veterinary immunology and immunopathology    February 9, 2006   Volume 110, Issue 3-4 363-367 doi: 10.1016/j.vetimm.2006.01.001
Wagner B, Hillegas JM, Antczak DF.Interleukin-4 (IL-4) is secreted by T helper type 2 cells, mast cells, basophils and eosinophils. Detection of IL-4 can contribute the evaluation of cellular immune responses during infectious diseases, immunological disorders or vaccination. We used recombinant equine IL-4 to generate a monoclonal antibody (mAb) to equine IL-4. The mAb detected recombinant IL-4 in mammalian cells transfected with different plasmids containing IL-4 cDNA. After mitogen stimulation of equine peripheral blood mononuclear cells, an intracellular protein was recognized by the new mAb in 1-2% of lymphocytes using fl...
Detection of the reemerging agent Burkholderia mallei in a recent outbreak of glanders in the United Arab Emirates by a newly developed fliP-based polymerase chain reaction assay.
Diagnostic microbiology and infectious disease    February 8, 2006   Volume 54, Issue 4 241-247 doi: 10.1016/j.diagmicrobio.2005.09.018
Scholz HC, Joseph M, Tomaso H, Al Dahouk S, Witte A, Kinne J, Hagen RM, Wernery R, Wernery U, Neubauer H.A polymerase chain reaction (PCR) assay targeting the flagellin P (fliP)-I S407A genomic region of Burkholderia mallei was developed for the specific detection of this organism in pure cultures and clinical samples from a recent outbreak of equine glanders. Primers deduced from the known fliP-IS407A sequence of B. mallei American Type Culture Collection (ATCC) 23344(T) allowed the specific amplification of a 989-bp fragment from each of the 20 B. mallei strains investigated, whereas other closely related organisms tested negative. The detection limit of the assay was 10 fg for purified DNA of ...
Sequence variation of the SeM gene of Streptococcus equi allows discrimination of the source of strangles outbreaks.
Journal of clinical microbiology    February 4, 2006   Volume 44, Issue 2 480-486 doi: 10.1128/JCM.44.2.480-486.2006
Kelly C, Bugg M, Robinson C, Mitchell Z, Davis-Poynter N, Newton JR, Jolley KA, Maiden MC, Waller AS.Improved understanding of the epidemiology of Streptococcus equi transmission requires sensitive and portable subtyping methods that can rationally discriminate between strains. S. equi is highly homogeneous and cannot be distinguished by multilocus enzyme electrophoretic or multilocus sequence-typing methods that utilize housekeeping genes. However, on sequence analysis of the N-terminal region of the SeM genes of 60 S. equi isolates from 27 strangles outbreaks, we identified 21 DNA codon changes. These resulted in the nonsynonymous substitution of 18 amino acids and allowed the assignment of...
Sensitization to horse allergens without apparent exposure to horse. Report of two cases.
European annals of allergy and clinical immunology    February 4, 2006   Volume 37, Issue 9 350-352 
Liccardi G, Dente B, Senna G, De Martino M, D'Amato L, D'Amato G.It has been well established that occupational exposure to horses increases the risk of allergic sensitization to horse hair, on the contrary very few data have been published in individuals without direct contact with horses. We describe two cases of severe respiratory symptoms triggered by a single direct exposure in two patients with only 4-5 previous short contacts with horses during the entire their life, since they live in urban area of Naples. Cutaneous and serological diagnostic tests showed significant sensitization to horse dander other than some common inhalant allergens. Our report...
Evaluation of concentration of voriconazole in aqueous humor after topical and oral administration in horses.
American journal of veterinary research    February 4, 2006   Volume 67, Issue 2 296-301 doi: 10.2460/ajvr.67.2.296
Clode AB, Davis JL, Salmon J, Michau TM, Gilger BC.To determine penetration of topically and orally administered voriconazole into ocular tissues and evaluate concentrations of the drug in blood and signs of toxicosis after topical application in horses. Methods: 11 healthy adult horses. Methods: Each eye in 6 horses was treated with a single concentration (0.5%, 1.0%, or 3.0%) of a topically administered voriconazole solution every 4 hours for 7 doses. Anterior chamber paracentesis was performed and plasma samples were collected after application of the final dose. Voriconazole concentrations in aqueous humor (AH) and plasma were measured via...
Evaluation of factors associated with positive IgM capture ELISA results in equids with clinical signs compatible with West Nile virus infection: 1,017 cases (2003).
Journal of the American Veterinary Medical Association    February 2, 2006   Volume 228, Issue 3 414-421 doi: 10.2460/javma.228.3.414
Tanner JM, Traub-Dargatz JL, Hill AE, Van Campen H, Knight AP, Cunningham WE, Salman MD.To describe the prevalence of West Nile virus (WNV) infection and evaluate factors associated with positive IgM capture ELISA results in equids with clinical signs compatible with WNV infection. Methods: Retrospective case series. Methods: Laboratory submission forms from 1,104 equids tested for WNV in Colorado in 2003. Methods: Submission forms accompanying samples submitted for detection of WNV via IgM capture ELISA were obtained from the Colorado state veterinarian and diagnostic laboratories performing the tests. Data on signalment, clinical signs, history of vaccination against WNV, and a...
How does magnetic resonance imaging represent histologic findings in the equine digit? Murray RC, Blunden TS, Schramme MC, Dyson SJ.Magnetic resonance (MR) imaging is increasingly used in the diagnosis of equine foot pain, but improved understanding of how MR images represent tissue-level changes in the equine foot is required. We hypothesized that alterations in signal intensity and tissue contour would represent changes in tissue structure detected using histologic evaluation. The study objectives were to determine the significance of MR signal alterations in feet from horses with and without lameness, by comparison with histopathologic changes. Fifty-one cadaver feet from horses with a history of lameness improved by pa...
Magnetic resonance imaging characteristics of the foot in horses with palmar foot pain and control horses. Murray RC, Schramme MC, Dyson SJ, Branch MV, Blunden TS.Palmar foot pain is a common cause of lameness. Magnetic resonance imaging (MRI) has the potential to detect damage in all tissues of the equine foot, but an understanding of the differences in magnetic resonance (MR) images between feet from horses with and without palmar foot pain is required. This study aimed to describe MR findings in feet from horses with no history of foot-related lameness, and to compare these with MR findings in horses with lameness improved by palmar digital local analgesia. Thirty-four limbs from horses euthanized with a clinical diagnosis of navicular syndrome (amen...
Intracranial abscess caused by Rhodococcus equi infection in a foal.
Journal of the American Veterinary Medical Association    January 24, 2006   Volume 228, Issue 2 251-253 doi: 10.2460/javma.228.2.251
Janicek JC, Kramer J, Coates JR, Lattimer JC, Lacarrubba AM, Messer NT.A 4-month-old Missouri Fox Trotter colt was examined for a 5-week history of head tilt after treatment for suspected pulmonary Rhodococcus equi infection. Results: Computed tomography revealed osteolysis of the occipital, temporal, and caudal portion of the parietal bones of the left side of the cranium. A soft tissue mass compressing the occipital region of the cerebral cortex and cerebellum was associated with the osteolytic bone. Results: A rostrotentorial-suboccipital craniectomy approach was performed to remove fragmented occipital bone, debulk the intracranial mass, and obtain tissue sam...
Effect of repeated transvaginal ultrasound-guided follicle aspiration on fertility in mares.
Journal of the American Veterinary Medical Association    January 24, 2006   Volume 228, Issue 2 248-250 doi: 10.2460/javma.228.2.248
Vanderwall DK, Hyde KJ, Woods GL.To determine whether performance of transvaginal ultrasound-guided follicle aspiration (TVUFA) repeatedly in mares adversely affects their fertility. Methods: Historical prospective study. Methods: 23 mares that had never undergone TVUFA and 59 mares that had undergone TVUFA on 1 to 11 occasions. Methods: Mares were classified into 4 groups according to the number of TVUFA procedures previously performed on the ovary in which ovulation occurred at the time of insemination as follows: group 1, 0 TVUFAs (control group, n = 23 mares); group 2, 1 or 2 TVUFAs (40 mare-cycles); group 3, 3 or 4 TVUFA...
Molecular diagnosis and equine parasitology.
Veterinary parasitology    January 20, 2006   Volume 136, Issue 2 109-116 doi: 10.1016/j.vetpar.2005.12.006
Hodgkinson JE.The future implementation of improved and sustainable control strategies for the major equine parasites will be dependent on a greater insight into their basic biology, pathogenicity and epidemiology together with an enhanced ability for accurate diagnosis. This paper will provide a review of the current molecular methods under development for the detection of equine parasites and their application to current scientific questions. In particular, the strongyles are recognised as important pathogens of horses and recent advances made in the study of this parasitic group at the single species lev...
Importance of using guarded techniques for the preparation of endometrial cytology smears in mares.
Theriogenology    January 20, 2006   Volume 66, Issue 2 423-430 doi: 10.1016/j.theriogenology.2005.12.007
Aguilar J, Hanks M, Shaw DJ, Else R, Watson E.Material for endometrial cytology can be collected by veterinarians using guarded or unguarded swabs, or digitally with a gloved hand, and is an important diagnostic tool in establishing the endometrial health of mares prior to breeding. The aim of this study was to determine whether the use of unguarded endometrial samples is a reliable indicator of the presence of neutrophils in the uterus. Duplicate endometrial smears were collected from 41 genitally normal, non-pregnant fertile mares by both double-guarded swabs (DGS) and in an unguarded manner by digital scraping (DS) of the endometrium. ...
The detection of anthelmintic resistance in nematodes of veterinary importance.
Veterinary parasitology    January 19, 2006   Volume 136, Issue 3-4 167-185 doi: 10.1016/j.vetpar.2005.11.019
Coles GC, Jackson F, Pomroy WE, Prichard RK, von Samson-Himmelstjerna G, Silvestre A, Taylor MA, Vercruysse J.Before revised World Association for the Advancement of Veterinary Parasitology (WAAVP) guidelines on the detection of anthelmintic resistance can be produced, validation of modified and new methods is required in laboratories in different parts of the world. There is a great need for improved methods of detection of anthelmintic resistance particularly for the detection of macrocyclic lactone resistance and for the detection of resistant nematodes in cattle. Therefore, revised and new methods are provided here for the detection of anthelmintic resistance in nematodes of ruminants, horses and ...
Real-time ultrasound measure of the fetal eye (vitreous body) for prediction of parturition date in small ponies.
Theriogenology    January 19, 2006   Volume 66, Issue 2 331-337 doi: 10.1016/j.theriogenology.2005.11.019
Turner RM, McDonnell SM, Feit EM, Grogan EH, Foglia R.Ultrasonographic fetal eye measures have been used to estimate gestational age of the fetus in light horse mares. However, fetal eye measures have not been published for smaller pony breeds. This study was conducted to develop reference ranges for ultrasonographic measures of fetal eyes in small ponies for the purpose of predicting days before parturition (DBP) when breeding or ovulation dates are unknown. Twenty-three Shetland-type pony mares were studied across one (n = 10) or two (n = 13) gestations in 2004 (18 pregnancies) and 2005 (18 pregnancies). Measurements of fetal eyes were obtained...
Effect of local analgesia on movement of the equine back.
Equine veterinary journal    January 18, 2006   Volume 38, Issue 1 65-69 doi: 10.2746/042516406775374351
Holm KR, Wennerstrand J, Lagerquist U, Eksell R, Johnston C.Diagnostic infiltration of local anaesthetic solution is commonly used in cases of equine back pain. Evaluation is subjective and it is not known how local analgesia of the back affects horses without clinical signs of back pain. Objective: To evaluate the effect of infiltration of local anaesthetics on the movement of the back in horses without clinical signs of back pain, and to evaluate the usefulness of kinematic studies as an objective and quantitative tool in evaluating local analgesia in clinical practice. Methods: The kinematics of the back in 10 clinically sound horses were measured o...
Constant rate infusion for topical ocular delivery in horses: a pilot study.
Veterinary ophthalmology    January 18, 2006   Volume 9, Issue 1 1-5 doi: 10.1111/j.1463-5224.2005.00424.x
Myrna KE, Herring IP.To assess the efficacy and kinetics of a continuous infusion modality for topical ophthalmic drug delivery via subpalpebral lavage in horses. Methods: Five ophthalmically normal horses Methods: A constant-rate continuous delivery pump was used to apply 0.01% fluorescein solution to the eyes of five ophthalmically normal horses via subpalpebral lavage. Fluorescein was applied at a rate of 0.14 mL/h for 72 h. Tear samples were collected from the ventral conjunctival fornix at regular intervals via capillary tube and tear fluorescein concentrations were determined via spectral assay. Laboratory s...
Qualitative and quantitative evaluation of equine respiratory mechanics by impulse oscillometry.
Equine veterinary journal    January 18, 2006   Volume 38, Issue 1 52-58 doi: 10.2746/042516406775374252
Van Erck E, Votion D, Art T, Lekeux P.The long-established conventional reference technique (CRT) for measuring respiratory mechanics in horses lacks sensitivity and there is a need for further refinement in new technology, such as the impulse oscillometry system (IOS). Objective: To evaluate the potential use of the IOS as a clinical respiratory function test and compare it to the current CRT in horses suffering from common upper and lower airway dysfunctions. Methods: Six healthy horses were tested before and after induction of a unilateral nasal obstruction (UNO) or transient left laryngeal hemiplegia (LLH). Six heaves-affected...
Molecular diagnosis of anthelmintic resistance.
Veterinary parasitology    January 18, 2006   Volume 136, Issue 2 99-107 doi: 10.1016/j.vetpar.2005.12.005
von Samson-Himmelstjerna G.Conventional and real time polymerase chain reaction-based tests have been developed for the diagnosis of anthelmintic resistance (AR) in populations of several small and large ruminant as well as horse gastro-intestinal nematode species. To date, molecular markers that correlate well with AR are available only for the detection of benzimidazole resistance. Recently, however, a single nucleotide polymorphism was found in vitro to be of functional relevance for reduced drug efficacy to macrocylic lactones. The focus of the present review, therefore, is the molecular mechanism of action of these...
Evaluation of an in vitro sulphidoleukotriene release test for diagnosis of insect bite hypersensitivity in horses.
Equine veterinary journal    January 18, 2006   Volume 38, Issue 1 40-46 doi: 10.2746/042516406775374333
Baselgia S, Doherr MG, Mellor P, Torsteinsdottir S, Jermann T, Zurbriggen A, Jungi T, Marti E.Insect bite hypersensitivity (IBH) is an IgE-mediated allergic dermatitis caused by bites of Culicoides and Simulium species, and improved means of diagnosis are required. Objective: The cellular antigen simulation test (CAST) with C. nubeculosus and S. vittatum extracts was assessed in a population of IBH-affected and healthy horses. Variations in test results over a one year period and possible cross-reactivity between different insect extracts was studied. Methods: A total of 314 mature horses were studied using the CAST. Influence of severity of clinical signs, gender and age were evaluate...
Evaluation of a nested PCR test and bacterial culture of swabs from the nasal passages and from abscesses in relation to diagnosis of Streptococcus equi infection (strangles).
Equine veterinary journal    January 18, 2006   Volume 38, Issue 1 59-63 doi: 10.2746/042516406775374324
Grønbaek LM, Angen O, Vigre H, Olsen SN.Streptococcus equi is the cause of strangles in horses. To improve diagnostic sensitivity, development and evaluation of DNA-based methods are necessary. Objective: To evaluate diagnostic methods and observe the pattern of bacterial shedding during natural outbreaks. Methods: Two herds with natural outbreaks of strangles were visited over a period of 15 weeks and 323 samples originating from 35 horses investigated. The diagnostic use of a nested PCR test was evaluated using a collection of 165 isolates of Lancefield group C streptococci (species specificity) and swabs from nasal passages or fr...
Mechanical nociceptive thresholds in the axial skeleton of horses.
Equine veterinary journal    January 18, 2006   Volume 38, Issue 1 70-75 doi: 10.2746/042516406775374315
Haussler KK, Erb HN.An objective measure of neck, back and croup pain is needed in horses with musculoskeletal injuries, vague upper limb lameness or poor performance. Objective: To establish mechanical nociceptive thresholds (MNTs) within the axial skeleton and evaluate the effects of subject status and ridden exercise. Methods: Thirty-six mature horses (10 nonridden; 26 actively ridden) were used to assess MNTs evoked by a pressure algometer (PA) with a 1 cm2 rubber plunger tip at 62 midline and bilaterally symmetrical anatomical landmarks along the axial skeleton. Pressure was applied at a uniform rate of appl...
Arthroscopic diagnosis and treatment of intra-articular insertional injuries of the suspensory ligament branches in 18 horses.
Equine veterinary journal    January 18, 2006   Volume 38, Issue 1 10-14 doi: 10.2746/042516406775374243
Minshall GJ, Wright IM.Clinical association between the branches of insertion of the suspensory ligament (SL) and metacarpophalangeal (MCP) and metatarsophalangeal (MTP) joints has been reported. However, there has been no assessment of the lengths of the SL branches which are subsynovial with respect to the joints or reports of involvement of the MCP/MTP joints in injuries of the SL branches. Objective: To establish proportions of SL branches subsynovial with respect to the MCP/MTP joints and report clinical and arthroscopic findings in horses with desmitis of SL branches identified as having an articular component...
Use of scintigraphy for diagnosis of equine paranasal sinus disorders.
Veterinary surgery : VS    January 18, 2006   Volume 35, Issue 1 94-101 doi: 10.1111/j.1532-950X.2005.00118.x
Barakzai S, Tremaine H, Dixon P.To describe and analyze subjective and objective scintigraphic findings in horses with paranasal sinus disorders. Methods: Horses with suspected disorders of the paranasal sinuses or cheek teeth (n = 48) and control horses (n = 30). Methods: 99Technetium-methylenediphosphate (99Tc-MDP; 7500 MBq) was administered for scintigraphic examination of the skull. Abnormal patterns of increased radionuclide uptake (IRU) were identified and subjectively described. Scintigrams and radiographs were blindly assessed by 2 clinicians and the accuracy of the imaging modalities was compared. Objective analysis...
Pressure algometry for the detection of induced back pain in horses: a preliminary study.
Equine veterinary journal    January 18, 2006   Volume 38, Issue 1 76-81 doi: 10.2746/042516406775374225
Haussler KK, Erb HN.Pressure algometry (PA) is a potential modality for objectively measuring mechanical nociceptive thresholds (MNTs) in horses. Its ability to differentiate musculoskeletal pain sites from nonpain sites is unknown and must be assessed prior to its clinical application. Objective: To assess the ability of PA to detect induced musculoskeletal pain. Methods: Twenty clinically normal mature horses (2 groups of 10 horses, over 2 years) were used to measure MNTs before and after implantation and removal of 2 fixation half-pins in the dorsal spinous processes of 2 adjacent vertebrae. To assess hyperalg...
Equine Culicoides hypersensitivity: evaluation of a skin test and of humoral response.
Journal of veterinary medicine. A, Physiology, pathology, clinical medicine    January 18, 2006   Volume 53, Issue 1 30-33 doi: 10.1111/j.1439-0442.2006.00783.x
Ferroglio E, Pregel P, Accossato A, Taricco I, Bollo E, Rossi L, Trisciuoglio A.Intradermal tests were carried out on 18 horses with clinical signs of Culicoides hypersensitivity (CHS) and 23 horses without clinical signs of CHS, and sera from these horses were analysed by SDS-PAGE and Western blotting (W-B). Intradermal injections of 0.1 ml of 25 microg/microl sterile Culicoides extract, 0.1 ml of 1:10,000 histamine (positive control) and 0.1 ml of physiological saline (negative control) were made in the dermis of the middle region of the neck. Analysis of reactions indicated that a 1 cm wheal and a skinfold thickness >10% at 24 h represented a valid cut-off between h...