Rapid Slide Agglutination Test Research Articles

Sero-epidemiological survey and risk factors associated with brucellosis in dogs in south-western Nigeria.

IntroductionIn Nigeria, there is limited information on brucellosis particularly in dogs, despite its public health implications. We undertook a sero-epidemiological survey of brucellosis in dogs to determine the prevalence of the disease and associated risk factors for its occurrence in Nigeria.MethodsA cross-sectional study was conducted to screen dogs in south-western Nigeria for antibodies to Brucella sp using the rapid slide agglutination test (RSA) and Rose Bengal test (RBT), with positive samples confirmed respectively by serum agglutination test (SAT) and competitive enzyme linked immunosorbent assay (cELISA). Data were analyzed with STATA-12.ResultsFrom the 739 dog sera tested, 81 (10.96%) were positive by RSA and 94 (12.72%) by RBT; these were corroborated with SAT (4/81; 4.94%) and cELISA (1/94; 1.06%), respectively. Logistic regression identified location (OR=0.04; 95% CI: 0.02-0.09), breed (OR=1.71; 95% CI: 1.34-2.19), age (OR=0.10; 95% CI: 0.04-0.30) and management system (OR=8.51; 95% CI: 1.07-68.05) as risk factors for Brucella infection by RSA. However, location (OR=10.83; 95% CI: 5.48-21.39) and history of infertility (OR=2.62; 95% CI: 1.41-4.84) were identified as risk factors using RBT.ConclusionGiven the 10.96% to 12.72% seroprevalence of brucellosis recorded in this study, we advocate control of the disease in dogs, and public health education for those at risk of infection. Again, further studies are required to elucidate the role of dogs in the epidemiology of brucellosis in Nigeria considering the conducive human-animal interface and ecological factors responsible for the transmission of the disease.

Usefulness of Urine as a Sample for Detection of Brucella Spp in Male Canines

Usefulness of Urine as a Sample for Detection of Brucella Spp in Male Canines Urine was used as a sample and Sensitivity (S), Specificity (Sp) and the positive Likelihood Ratio (LR+) of molecular and serological methods, combined with epidemiology and the clinical symptoms for detection of Brucella spp., were compared in blood and urine samples from 241 male canines. The rapid slide agglutination test together with 2- mercaptoethanol (2-ME RSAT) were used as a screening test, followed by confirmation using an indirect immunoenzymatic assay (iELISA) and bacteriological culture. Results were as follows: Test a) PCR (Polymerase Chain Reaction) of blood compared to blood culture: S 80%, Sp 92%, LR+ 10.32% (CI 5.27-19.20) test b) iELISA compared to blood culture: S 100%, Sp 94%, LR+: 16.57%) (CI 9.97-27.53), test c) PCR of urine compared to urine culture: S 100%, Sp 93% (CI 8.36-21.56), LR+: 13.64 test d) iELISA compared to urine culture: S 100%, Sp 93%, RV+: 14.5 (CI 9.03-23.26). We conclude that molecular and serological tests in conjunction with epidemiology are both useful for diagnosis and that both blood and urine samples should be assayed together.

Serotyping reanalysis of unserotypable Actinobacillus pleuropneumoniae isolates by agar gel diffusion test.

We observed increasing unserotypable (UT) Actinobacillus pleuropneumoniae isolates using agar gel diffusion (AGD) test. To reanalyze their serovar, we performed rapid slide agglutination (RSA) test and multiplex PCR for 47 UT isolates. Of these, 25 were serovar 1 (UT-serovar 1), 20 were serovar 2 (UT-serovar 2) and 2 were serovar 15 (UT-serovar 15). We examined serotyping antigen extraction temperature to determine heat influence. UT-serovar 1 and 15 were influenced by heat, because their precipitation lines were observed in the case of low antigen extraction temperature. To investigate the relationship between antigenicity and genotype, we performed pulsed-field gel electrophoresis (PFGE) analysis using UT-serovar 2 and 15. The predominant PFGE pattern of UT-serovar 2 was identical to that of serovar 2.

Detection of Antibiotic Resistant Staphylococcus aureus from Milk: A Public Health Implication.

The aim of this study was to investigate the occurrence, antibiotic susceptibility profiles, and virulence genes determinants of S. aureus isolated from milk obtained from retail outlets of the North-West Province, South Africa. To achieve this, 200 samples of raw, bulk and pasteurised milk were obtained randomly from supermarkets, shops and some farms in the North-West Province between May 2012 and April 2013. S. aureus was isolated and positively identified using morphological (Gram staining), biochemical (DNase, catalase, haemolysis and rapid slide agglutination) tests, protein profile analysis (MALDI-TOF mass spectrometry) and molecular (nuc specific PCR) methods. The antimicrobial resistance profiles of the isolates were determined using the phenotypic agar diffusion method. Genes encoding enterotoxins, exfoliative toxins and collagen adhesins were also screened using PCR. Among all the samples examined, 30 of 40 raw milk samples (75%), 25 of 85 bulk milk samples (29%) and 10 of 75 pasteurised milk samples (13%) were positive for S. aureus. One hundred and fifty-six PCR-confirmed S. aureus isolates were obtained from 75 contaminated milk samples. A large proportion (60%–100%) of the isolates was resistant to penicillin G, ampicillin, oxacillin, vancomycin, teicoplanin and erythromycin. On the contrary, low level resistance (8.3%–40%) was observed for gentamicin, kanamycin and sulphamethoxazole. Methicillin resistance was detected in 59% of the multidrug resistant isolates and this was a cause for concern. However, only a small proportion (20.6%) of these isolates possessed PBP2a which codes for Methicillin resistance in S. aureus. In addition, 32.7% of isolates possessed the sec gene whereas the sea, seb sed, see, cna, eta, etb genes were not detected. The findings of this study showed that raw, bulk and pasteurised milk in the North-West Province is contaminated with toxigenic and multi-drug resistant S. aureus strains. There is a need to implement appropriate control measures to reduce contamination as well as the spread of virulent S. aureus strains and the burden of disease in humans.

Seroprevalence of Salmonella Gallinarum Infection in Chicken Population of parts of Tigray and Addis Ababa by Plate Agglutination and Micro-agglutination Tests

Ethiopia owns a large poultry population whose growth is highly constrained by diseases. Fowl typhoid is a serious concern in growing and adult poultry and results from infection by Salmonella Gallinarum (Salmonella enterica subsp. enterica serovar Gallinarum biovar Gallinarum). Knowledge of the prevalence of the disease with confirmatory diagnosis is of paramount importance to embark on a control or prevention program. Seroprevalence of this infection in chicken population of Ethiopia has been reported from some parts of this country but in most of the reports it is based exclusively on a rapid slide agglutination test without further confirmation by another specific test. This study screened 279 sera samples of poultry for S. Gallinarum infection by employing plate agglutination test using colored antigen of S. Gallinarum that revealed 125(44.80%) of 279 sera samples positive. However, when plate positive samples were further tested for confirmation by microagglutination test by employing plain antigen of S. Gallinarum, only 55 (19.71%) were serologically positive lowering the prevalence rate from 44.8% to 19, 71%. The study reveals the serological prevalence of Salmonella gallinarum infection in poultry population of the study areas and recommends that the initial screening of this infection by a test like plate agglutination must always be followed by another specific test to get an accurate estimate of this infection. Keywords : Micro-agglutination test, Plate agglutination test, Poultry population, Salmonella gallinarum, Seroprevalence, Addis Ababa, Tigray, Ethiopia.

육용종계 가금티프스 발생에 따른 검사방법 개선 방안

Pullorum disease and fowl typhoid are septicemic avian diseases transmitting through egg by transovarian infections. A series of tests has been performing in breeding flocks of chickens and test plans for proper inspection have been modified by government veterinary institute to control of such diseases. To improve inspection plans, different test methods were compared using fowl typhoid positive samples from a poultry farm located in Jeonbuk state in 2012. Based on first inspection, 11 samples among total 200 samples were positive by rapid slide agglutination (RSA) test and 7 samples among RSA positive samples were finally diagnosed as Salmonella Gallinarum infection by ELISA, bacterial isolation, PCR, and histopathologic examination. In the second inspection, 20 samples among total 100 samples were positive by RSA test. Among RSA positive ones, 19 samples were positive by ELISA, S. Gallinarm were successfully isolated in 3 samples, and 16 samples were positive by PCR in the cecal tonsils where were not successful for bacterial isolation. Based on histopathologic examination, severe inflammation in the 13 cecal tonsils and infiltration of lymphocytes and heterophils in the 11 livers were observed. Therefore, we suggest that bacterial isolation, PCR, and histopathologic examination methods in the third inspection need to be further used in various tissues for correct diagnosis and for final eradication of pullorum disease and fowl typhoid in breeding flocks of chickens.

Evidence for unapparent Brucella canis infections among adults with occupational exposure to dogs.

Human serological assays designed to detect brucellosis will miss infections caused by Brucella canis, and low levels of periodic bacteremia limit diagnosis by blood culture. Recent B. canis outbreaks in dogs and concomitant illnesses in caretakers suggest that unapparent human infections may be occurring. With more than a quarter of a million persons in occupations involving dogs, and nearly 80 million dog owners in the United States, this pathogen is an under-recognized human health threat. To investigate occupational exposure to B. canis, we adapted a commercial canine serological assay and present the first controlled seroepidemiological study of human B. canis infections in recent years. 306 adults with occupational exposure to dogs and 101 non-matched, non-canine-exposed subjects were enrolled. Antibodies were detected using the canine D-Tec(®) CB rapid slide agglutination test (RSAT) kit with a secondary 2-mercaptoethanol (ME)-RSAT. Results were validated on a blinded subset of sera with an additional RSAT and indirect enzyme-linked immunoassay at the National Administration of Laboratories and Health Institutes (ANLIS) in Argentina. Seroprevalence ranged from 10.8% (RSAT) to 3.6% (ME-RSAT) among canine-exposed subjects. Kennel employees were more likely to test RSAT seropositive compared with other canine exposures (OR = 2.7; 95% CI, 1.3-5.8); however, low seroprevalence limited meaningful occupational risk factor analyses. Two seropositive participants reported experiencing symptoms consistent with brucellosis and having exposure to B. canis-infected dogs; however, temporality of symptom onset with reported exposure could not be determined. D-Tec(®) CB results had substantial agreement with ANLIS assays (Cohen's kappa = 0.60-0.68). These data add to a growing body of literature suggesting that people occupationally exposed to dogs may be at risk of unapparent B. canis infection. It seems prudent to consider B. canis as an occupational public health concern and encourage the development of serological assays to detect human B. canis infections.

Brucella canis’in Serolojik Tanısında Kullanılan Çabuk Lam Aglütinasyon Test (ÇLAT) Antijeninin Fermentörde Farklı Besiyerlerinde Karşılaştırmalı Olarak Üretimi

Brucella canis’in Serolojik Tanısında Kullanılan Çabuk Lam Aglütinasyon Test (ÇLAT) Antijeninin Fermentörde Farklı Besiyerlerinde Karşılaştırmalı Olarak Üretimi

Diagnostic efficacy of Widal slide agglutination test against Widal tube agglutination test in enteric fever

Introduction: Enteric fever is an endemic disease in India and warrants rapid and affordable diagnosis. The Widal slide agglutination test is a commonly used rapid screening test for this purpose. The literature available on its diagnostic ability in comparison to the tube agglutination test is however scanty. Hence, this study aims to evaluate the efficacy of the Widal slide agglutination test and the tube agglutination test for the diagnosis of enteric fever. Materials and Methods: A total of 1470 sera were collected during the study period of one year from patients having pyrexia of unknown origin. All the samples were tested for the presence of anti O and anti H agglutinins against S. typhi and S. paratyphi A by semi quantitative slide and quantitative tube agglutination tests as per standard protocols. The titers of 1:80 (O agglutinins) and 1:160 (H agglutinins) were taken as the significant titer for the diagnosis of enteric fever. The results of the slide agglutination test were compared with the tube agglutination test and analyzed using Fisher's exact test. The sensitivity, specificity, positive and negative predictive values of slide agglutination were calculated using the tube agglutination method as a standard for comparison. Results: Of the 294 slide positive samples, 209 (71.1%) samples tested negative by the tube agglutination test. The sensitivity, specificity, positive and negative predictive values for the slide agglutination test were observed to be 100 % (CI 95.75-100%), 84.91% (CI 82.93-86.73%), 28.91% (CI 23.84-34.45%) and 100% (CI 99.69-100%) respectively. Conclusion: Serological diagnosis of enteric fever should always be confirmed by the tube agglutination test rather than depending solely upon the rapid slide agglutination test results.

Unusual extraocular manifestations in moraxellosis of a West African Dwarf goat: a case report

A 4-year-old, 14.5-kg doe with primary complaint of abortion and blindness was presented to the University of Nigeria, Nsukka Veterinary Teaching Hospital (UNVTH). Clinical examination revealed ocular (bilateral keratoconjunctivitis, corneal opacity, mucopurulent ocular discharge and blindness) and oral (erosive stomatitis, lingual vesicles and dental attrition) lesions. Neither intracellular parasites nor inclusion bodies were found in the stained impression smears. Rapid slide agglutination test (RSAT) was negative for brucellosis. Haematology showed normal packed cell volume (PCV), while serum biochemical assay revealed normal alanine aminotransferase (ALT), slightly increased aspartate aminotransferase (AST) and markedly increased alkaline phophatase (ALP). Diagnosis of moraxellosis was based on isolation and identification of pure cultures of beta-haemolytic Moraxella bovis strain from cultured samples following standard protocols. Antibiotic sensitivity result showed that the isolate was resistant to chloramphenicol, ciprofloxacin, ampicillin and perfloxacin, but susceptible to gentamicin, orfloxacin and tetracycline. Intramuscular injection of 5 % oxytetracycline short-acting and multivitamin for 5 days was very effective. This report indicates that extraocular manifestations could possibly occur in caprine moraxellosis.

Prevalence of Toxoplasma gondii and Associated Risk Factors among People Living with HIV at Gondar University Hospital, Northwest Ethiopia

Background. Toxoplasma gondii is an obligate intracellular protozoan parasite and is a major opportunistic pathogen in immune-compromised hosts. This study assessed the prevalence of T. gondii and associated risk factors among people living with HIV. Methods and Materials. A cross-sectional study was carried out among people living with HIV attending Gondar University Hospital. A structured and pretested questionnaire was used to collect sociodemographic factors, and 10mL of venous blood was collected for anti-Toxoplasma antibody test and determination of CD4 levels. Serum was tested in duplicate for anti-Toxoplasma antibody using rapid slide agglutination test. Results. A total of 170 study subjects were enrolled in the study. Seroprevalence of T. gondii among the study participants was 76.5% (95% CI: 69.0–82.8). High proportions of seropositive individuals (64.7%) were found under the child bearing age groups. The mean CD4+ lymphocyte count of HIV monoinfected participants was cells/mm3 while coinfected study participants had mean CD4+ lymphocyte count of cells/mm3 with value of 0.01. Conclusion. The seroprevalence of T. gondii among people living with HIV was high. Cautious followup of HIV-positive patients is needed to prevent development of toxoplasmic encephalitis and other related complications.

Exposure to <em>Brucella canis</em> in Owned and Stray Dogs in Grenada, West Indies

The aim of this cross sectional study was to estimate the prevalence of exposure to <em>Brucella canis</em> in owned and stray dogs in Grenada. Documentation of exposure to this important bacterial pathogen in dogs will facilitate instituting appropriate prevention and control measures and education of the public about potential zoonotic implications. Serum samples were collected from 255 stray and 204 owned dogs in Grenada from 2009 to 2011. Sera were initially screened with the rapid slide agglutination test and the positive samples further tested with 2-mercaptoethanol rapid slide agglutination test. Exposure to <em>B. canis</em> was found in 10 stray dogs but none in owned dogs.

Investigation of Brucella canis Seroprevalence in Brucellosis Suspected Cases

Brucella canis which is the main etiologic agent of brucellosis in dogs, can be transmitted to man. It causes mild or asymptomatic infection in human compared with other Brucella species. B.canis can be transmitted to man either by laboratory accidents or contact with infected dogs. Since B.canis infections in humans are not routinely investigated in hospitals in Turkey, the data are limited to reveal the current status of B.canis infections in people in our country. The purpose of this study was to determine the seroprevalence of B.canis infection in brucellosis-suspected cases. The study was conducted at Konya Education and Research Hospital, (located at Central Anatolia of Turkey) during March-August 2010 period. Serum samples were obtained from 1000 patients (age range: 15-65 years; 652 of them were women) presented with brucellosis-like symptoms, including fever, headache, night sweats, appetite loss, weakness, arthralgia and myalgia. Rose Bengal Plate Tests (Seromed, Turkey) for smooth Brucella species were negative in all serum samples. Rough type B.canis antigen was prepared with B.canis NCTC 10854 strain for serodiagnosis. Antibody responses to B.canis in the serum samples were investigated by rapid slide agglutination test (SAT) and modified plate agglutination test (MPAT). Of the 1000 sera tested, 34 (0.34%) were found to be positive with SAT while the remaining were found negative. MPAT was used for the detection of antibody titer and 22 (0.22%) out of 1000 sera were found positive with MPAT (one had 1/48, five had 1/96, six had 1/192, six had 1/384, four had 1/768 titers). Among 22 positive patients, 17 were female and five were male, and the difference between the genders was found statistically significant (p< 0.05). It was concluded the use of both S and R antigens in the serological tests applied for the diagnosis of brucellosis in our country will supplement both diagnosis and seroepidemiological data related to brucellosis.

Preliminary Study of an Immunochromatography Test for Serological Diagnosis of Canine Brucellosis

The most widely used screening test for the diagnosis of brucellosis in the dog is the rapid slide agglutination test in the presence of 2-mercaptoethanol (2ME-RSAT). The diagnosis is partially confirmed by the agar gel immunodiffusion test (AGID) and definitively confirmed by bacteriological isolation. Some chronic cases not detected by these tests may be detected by ELISA tests. The use of 2ME-RSAT in routine clinical practice requires a microscope and an experienced operator. An immunochromatographic diagnostic test for canine brucellosis (FASTest(®) Brucella c., Megacor, Hörbranz, Austria) has been recently released. In this study, we compared the diagnostic performance of the FASTest with those of 2ME-RSAT, AGID and ELISAs. Sera from 17 healthy dogs used as negative controls yielded negative results by FASTest, indicating a 100% specificity in this sample. Among 27 sera of dogs with acute or subacute brucellosis confirmed by B. canis isolation, all of which were positive by RSAT and ELISAs, the FASTest was positive in 24 cases and AGID in 23. In acute and subacute cases, the sensitivity of FASTest was 89%. Sera from six dogs with bacteriologically confirmed chronic brucellosis, which were positive by ELISAs but negative by 2ME-RSAT, were also tested; 1 was positive by FASTest and 4 were positive by AGID. These preliminary results indicate a good specificity of the FASTest (100% in this sample) but an unacceptable sensitivity as a screening test. In cases with chronic brucellosis, the sensitivity of the FASTest was lower than that of ELISAs but this assay could make a good intermediate test to be run after a positive RSAT and before running an AGID.

Oxidant and antioxidant parameters in patients with Brucella canis

The aim of this study was to evaluate the oxidative status in patient with Brucella canis (B. canis) and healthy controls by measurement of total antioxidant capacity (TAC) and total oxidant capacity (TOS), and calculation of oxidative stress index (OSI). Six patients with B. canis and 6 healthy control subjects were included in this study. Diagnosis of B. canis was made by rapid slide agglutination test (RSAT) and Modified Plate Agglutination Test (MPAT). The serum samples of B. canis positive patients and controls were used for measurement of TAC and TOS levels. The TOS and OSI index were significantly higher in brucellosis positive patients than control group (p<0.001 and p<0.0001, respectively), while TAC level was found similar in patient and control groups. Findings of this study have shown that B. canis generate low TOS and OSI index in man compared with smooth Brucella species infection. This difference may help to provide understanding about the pathogenesis of B. canis infection in man.

Assessing antibiotic resistance profiles in Escherichia coli and Salmonella species from groundwater in the Mafikeng area, South Africa

Escherichia coli and Salmonella species occur as normal flora in the gastrointestinal tract of animals and humans. However, pathogenic strains exist that cause disease in humans. Infections may result from the consumption of water and food contaminated with faeces of human and animal origin. In South Africa, residents of most rural communities rely on untreated ground water for survival. This practice results to the transfer of pathogenic micro-organism to humans and thus amplifies the need to identify contaminated water systems. Results obtained may adequately address water quality problems and hence protect public health. The study was conducted to isolate and determine the antibiotic resistance profiles of E. coli and Salmonella species from the groundwater obtained from two rural communities in the North-West province, S.A. Nineteen ground water samples were analyzed for characters of E. coli and Salmonella species. Only those isolates that satisfied all the primary (oxidase and the triple sugar iron) and secondary identification criteria (API 20E and rapid slide agglutination test) for E. coli and Salmonella species were used. A total of 63 E. coli and 64 Salmonella isolates were identified. The antibiotic susceptibilities of these isolates were evaluated against a panel of 10 antibiotics. A large proportion (56.7 to 57.6%) of the E. coli isolated from both Dibate and Verdwall were resistant to vancomycin. Similarly, large proportions (51.5 to 78.8%) of the E. coli isolated from Verdwall were resistant to ampicilin and erythromycin when compared to those isolated from Dibate. Furthermore, a higher percentage (90.0 to 100%) of Salmonellaisolated from both Dibate and Verdwall were resistant to ampicillin, erythromycin and vancomycin. AP-E-VA was the predominant phenotype for E. coli isolated from both Dibate and Verdwall in 40 and 20%, respectively, while the phenotypes AP-E-T-VA and AP-E-VA were dominant among the Salmonella species isolated in this study. The results indicated that E. coli and Salmonella could serve as indicator organisms necessary to assess the quality of ground water and their levels are critical parameters that could help to drive management strategies. This will limit the effect of these pathogens on consumers, as some water samples were visibly identified to be contaminated. Key words: Salmonella, E. coli, multiple antibiotic resistant (MAR), phenotype.

A Serological Diagnostic Survey for &lt;i&gt;Brucella canis&lt;/i&gt; Infection in Turkish Patients with Brucellosis-Like Symptoms

The incidence of Brucella canis infection in humans is unknown in Turkey. In this study, we investigated the prevalence of B. canis infection in human sera obtained from six regions in Turkey and comparatively evaluated the results obtained by agglutination-based techniques using standardized antigens made from B. canis. The patients (n = 1,746) presented with clinical symptoms that were similar to those of brucellosis. All patients who tested negative in the Rose Bengal test for the smooth Brucella strains (abortus, melitensis, and suis) were screened for evidence of B. canis infection using the rapid slide agglutination test (RSAT), the microagglutination test (MAT), and the 2-mercaptoethanol RSAT test (2ME-RSAT). Of the samples tested, 157 (8.9%), 68 (3.8%), and 66 (3.7%) were positive for B. canis, as determined by RSAT, MAT, and 2ME-RSAT, respectively. The diagnostic sensitivity, specificity, positive predictive value, and negative predictive value of RSAT were 100%, 94.6%, 42%, and 100%, respectively, and of MAT were 100%, 99.9%, 97%, and 100%, respectively. We recommend the routine use of MAT and 2ME-RSAT to check the sera of all patients with symptoms of brucellosis who are negative for brucellosis using a smooth Brucella antigen.

Canine brucellosis 검출을 위한 ELISA 진단법 확립

This study was carried out to develope enzyme-linked immunosorbent assay (ELISA) for detection of canine brucellosis in dogs experimentally inoculated with Brucella abortus 1119-3 and B. canis RM666. Groups A, B and C of dogs (each group consisting of three dogs) were orally inoculated with approximately 5×10 9 colony-forming units of B. abortus and B. canis, and with sterile pyrogen-free PBS, respectively. The animals were monitored at regular intervals upto the 12th week post inoculation (PI) by standard tube agglutination test (STAT), plate agglutination test (PAT), Rose Bengal test (RBT), 2-mercaptoethanol rapid slide agglutination test (2ME-RSAT) and ELISA. The induced antibody titers in group A dogs were detected from the first week PI to the eighth week PI in STAT, PAT and RBT using the inactivated whole cells of B. abortus 1119-3 as antigens, while no sera in groups B and C dogs reacted with the antigens. In 2ME-RSAT using whole cells of B. canis M-strain as antigens, the induced antibody titers in group B dogs were observed at the second week PI and persisted for the 12th week PI, while sera of groups A and C dogs did not react with the whole cells. In ELISA using cytoplasmic fractions antigen of B. abortus 1119-3, the mean optical density of antibodies in groups A and B was detected from the first and second weeks PI, respectively, and persisted for 12th week PI, while sera of group C did not cross-react with the fractions antigen. However, in ELISA using the hot saline extracts of B. canis M- as an antigen, the induced antibody titers in only group B dogs were detected from second week PI and persisted for until the end of this study. These results indicate that the ELISA using B. abortus 1119-3 cytoplasmic fractions as antigens can be a good candidate for detection of brucellosis by B. abortus as well as B. canis in dogs.

Seroprevalence of Brucella abortus and B. canis in household dogs in southwestern Nigeria: a preliminary report

A preliminary serological study of 366 household dogs in Lagos and Ibadan, southwestern Nigeria, was carried out to determine antibodies due to exposure to Brucella abortus and B. canis, using the rose bengal test (RBT) and the rapid slide agglutination (RSA) test, respectively. Results showed that 5.46 % (20/366) and 0.27 % (1/366) of the dogs screened were seropositive to B. abortus and B. canis, respectively. Of all dogs, 36 had a history of being fed foetuses from cows and 11 (30.6 %) of these tested positive in the RBT. Our findings, although based on a limited sample size and a dearth of clinical details, revealed that dogs in Nigeria may be infected with Brucella spp. given the wide range of risk factors. Further studies are recommended to elucidate the epidemiology of brucellosis in dogs and its possible zoonotic consequences in the country.

Detection of Brucella Canis–Induced Reproductive Diseases in a Kennel

Brucella spp. were isolated from an abortion case submitted for laboratory examination 8 months after the first clinical symptoms appeared in a kennel consisting of 31 dogs. Pathological investigations revealed the parallel presence of necrotic placentitis and the strong immunostaining of trophoblast cells by immunohistochemistry (IHC) using hyperimmune rabbit anti-Brucella canis primary antibodies. The rapid slide agglutination test was positive in 7 of 31 (23%) cases. The organism B. canis was successfully cultured from the blood, tissues, or vaginal swabs of only 3 of 31 (10%) cases. The isolated strains were identified as B. canis based on their colony morphology and agglutination with R sera. The strains were initially misidentified as B. suis with the "Bruce-ladder" method, and were subsequently correctly identified as B. canis with a single nucleotide polymorphism (SNP) typing test. Three culture-positive cases and 3 culture-negative cases with histories of reproductive disorders were selected and examined for the presence of B. canis infection using histopathology, IHC, and polymerase chain reaction (PCR) assays. Characteristic histologic lesions were found in all of the 6 animals, whereas IHC and PCR yielded positive results only in single cases from both groups. The results imply that all cases of canine abortion should be examined for brucellosis by bacterial culture of aborted fetuses and placentas. Immunohistochemical examination of placentas is also recommended because it is a quick and sensitive technique compared with bacterial culture. Multiple methods (i.e., serology, blood, and genital bacterial cultures) should be applied simultaneously and repeatedly for the reliable screening of B. canis infection in live individuals.