Higher non-return rate associated with Mycobacterium avium subspecies paratuberculosis infection at early stage in Holstein dairy cows

The effects of infection by Mycobacterium avium paratuberculosis (Map) on dairy cows are poorly documented and quite controversial. This retrospective study aimed at quantifying the variation in non-return to service of Holstein dairy cows according to their Map-infection status. Three different statuses were defined based on both individual and herd tests results: ELISA positive cow, all tests negative cow in a negative herd and all tests negative cow in a positive herd. Whatever the age at Map testing, the status was attributed to a cow from its first lactation onwards. Non-return to service was determined at 200 days after first and second services. The study was performed from 1999 to 2007 on 185,950 AI from 48,914 cows in early stage of the infection in 1069 herds by logistic regression controlling for known factors influencing non-return rate. Non-return rate was higher for infected cows compared to negative cows from negative herds (RR of 1.10 or +3.9 points of % of non-return rate). The effect was significant for parities 1 and 2 (RR of 1.11 and 1.12, respectively) but not for higher ones. This effect was lower when comparing positive cows to negative cows in the same herds but relative risks were still above 1. The hypothesis that the effect of Map on non-return depends upon the stage of infection is formulated.     

Evaluation of bioelectronics sensor compared to other diagnostic test in diagnosis of Johne's disease in goats

A bioelectronics sensor has been developed and it is evaluated for the diagnosis of paratuberculosis in goats. Initially hematite nanoparticles were prepared and using this nanoparticles as core, electrically active polyaniline coated magnetic (EAPM) nanoparticles are synthesized from aniline monomer (made electrically active by acid doping). These EAPM nanoparticles were fabricated with rabbit anti-goat IgG for the detection of goat antibodies on the capture pad. The protoplasmic antigen of Mycobacterium avium subspecies paratuberculosis (MAP) immobilized onto the capture pad will detect the antibody against MAP in the goat sera samples. This bound goat antibody will be detected by the anti-goat IgG previously bound to EAPM. Upon detection the EAPM nanoparticles bridges an electric circuit between the silver electrodes, flanking the capture membrane. The electrical conductance, caused by EAPM, was measured as direct charge transfer between the electrodes. Testing of the biosensor with known Johne's disease (JD) positive and negative serum samples gave significant difference in the electrical conductance value. Further the efficacy of this biosensor was compared with other serological tests like agar gel immunodiffusion (AGID) and absorbed ELISA using field sera. Out of 265 goat sera tested, positive results recorded were; AGID 36 (13.59%), bioelectronics sensor 49 (19.14%), and absorbed ELISA 51 (19.25%). This biosensor was also compared in live animals using intradermal Johnin test and nested PCR (detecting mycobacterial DNA in feces) in 65 animals. Of which, positive results recorded in animals were; Johnin test 21 (32%), biosensor 26 (40%) and fecal PCR detected mycobacterial DNA in 28 (43%) animals. Though the nanobioelectronics sensor was slightly less sensitive (not statistically significant) compared to absorbed ELISA and fecal nested PCR for mycobacterial DNA but it was simple to perform in field conditions and requires less time. The speed of detection and the equipment involved would support its application toward the various point-of-care opportunities aimed at control and management of Johne's disease in goats.     

Agar gel immunodiffusion test (AGID) evaluation for detection of bovine paratuberculosis in Rio de Janeiro,Brazil

AIMS: The aim of this study is to evaluate the AGID serological test for detection of antibodies anti-Mycobacterium paratuberculosis and its possible adoption as diagnostic method in our field conditions. METHODS: Bovine serum samples from dairy herds in Rio de Janeiro State, Brazil, were screened for the presence of antibodies against Myco. paratuberculosis using three different ELISA tests. A panel of 48 randomly selected sera were evaluated by an Agarose Gel Immunodiffusion (AGID) test using Protoplasmatic Paratuberculosis Antigen (PPA). AGID results were compared to the standards--the results of the three ELISA tests, and the specificity and sensitivity were calculated. RESULTS: From 48 sera tested for AGID, 14 (29.17%) were positive and 34 (70.83%) were negative. AGID sensitivity was 57% with two false-positive reactions, and specificity was 92.5% with nine false-negative results. The positive predictive value was calculated in 85.7% for a confidence interval of 95%. SIGNIFICANCE AND IMPACT OF THE STUDY: Due to its low sensitivity and specificity rates, AGID test has shown to be unsatisfactory as a screening diagnostic method for subclinical herd infection, but it can be useful as a confirmatory test for clinical suspect animals.     

Coxiella burnetii Shedding Routes and Antibody Response after Outbreaks of Q Fever-Induced Abortion in Dairy Goat Herds

Q fever is a zoonosis caused by Coxiella burnetii, a bacterium largely carried by ruminants and shed into milk, vaginal mucus, and feces. The main potential hazard to humans and animals is due to shedding of bacteria that can then persist in the environment and be aerosolized. The purpose of this study was to evaluate shedding after an outbreak of Q fever abortion in goat herds and to assess the relationship with the occurrence of abortions and antibody responses. Aborting and nonaborting goats were monitored by PCR for C. burnetii shedding 15 and 30 days after the abortion episodes. PCR analysis of all samples showed that 70% (n = 50) of the aborting and 53% (n = 70) of the nonaborting goats were positive. C. burnetii was shed into vaginal mucus, feces, and milk of 44%, 21%, and 38%, respectively, of goats that aborted and 27%, 20%, and 31%, respectively, of goats that delivered normally. Statistical comparison of these shedding results did not reveal any difference between these two groups. PCR results obtained for the vaginal and fecal routes were concordant in 81% of cases, whereas those for milk correlated with only 49% of cases with either vaginal or fecal shedding status. Serological analysis, using enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay (IFA), and complement fixation tests, showed that at least 24% of the seronegative goats shed bacteria. Positive vaginal and fecal shedding, unlike positive milk shedding, was observed more often in animals that were weakly positive or negative by ELISA or IFA. Two opposite shedding trends were thus apparent for the milk and vaginal-fecal routes. Moreover, this study showed that a nonnegligible proportion of seronegative animals that delivered normally could excrete C. burnetii.     

Mycobacterium Avium subsp. Paratuberculosis Isolates Induce In Vitro Granuloma Formation and Show Successful Survival Phenotype,Common Anti-Inflammatory and Antiapoptotic Responses within Ovine Macrophages Regardless of Genotype or Host of Origin

The analysis of the early macrophage responses, including bacterial growth within macrophages, represents a powerful tool to characterize the virulence of clinical isolates of Mycobcaterium avium susbp. paratuberculosis (Map). The present study represents the first assessment of the intracellular behaviour in ovine monocyte-derived macrophages (MDMs) of Map isolates representing distinct genotypes (C, S and B), and isolated from cattle, sheep, goat, fallow deer, deer, and wild boar. Intracellular growth and survival of the selected isolates in ovine MDMs was assessed by quantification of CFUs inside of the host cells at 2 h p.i. (day 0) and 7 d p. i. using an automatic liquid culture system (Bactec MGIT 960). Variations in bacterial counts over 7 days from the baseline were small, in a range between 1.63 to 1.05-fold. After 7 d of infection, variations in the estimated log₁₀ CFUs between all the tested isolates were not statistically significant. In addition, ovine MDMs exhibited enhanced anti-inflammatory, antiapoptotic and antidestructive responses when infected with two ovine isolates of distinct genotype (C and S) or with two C-type isolates from distinct hosts (cattle and sheep); which correlated with the successful survival of these isolates within ovine MDMs. A second objective was to study, based on an in vitro granuloma model, latter stages of the infection by investigating the capacity of two Map isolates from cattle and sheep to trigger formation of microgranulomas. Upon 10 d p.i., both Map isolates were able to induce the formation of granulomas comparable to the granulomas observed in clinical specimens with respect to the cellular components involved. In summary, our results demonstrated that Map isolates from cattle, sheep, goats, deer, fallow-deer and wild boar were able not only to initiate but also to establish a successful infection in ovine macrophages regardless of genotype.     

Prevalence of coccidia and gastrointestinal nematode infections in cross bred goats in the dry areas of Sri Lanka

The prevalence and intensity of oocysts of coccidia, particularly Eimeria species, and eggs of gastrointestinal nematodes in kids (2–4 months), young goats (5–12 months) and adult goats (>1 year) were determined in five large herds managed extensively in five villages in the dry zone. Of the representative samples examined, oocysts were found in 88% of kids, 91% of young goats and 83% of adults. Seven species of Eimeria were identified in faecal samples by salt flotation. Eimeria ninakohlyakimovae (31%), E. alijevi (29%) and E. arloingi (21%) were the most common species in all age categories. Other species encountered were E. christenseni (7%), E. jolchijevi (6%), E. hirchi (3%) and E. aspheronica (1%). Gastrointestinal nematode eggs were found in 89% of the kids, 94% of the young goats and 84% of the adult goats. Identification of gastrointestinal nematodes revealed Haemonchus contortus (90%) followed by Oesophogostomum spp. (8.5%) and Trichostrongylus spp. (1%) in all three age categories examined. Both Eimeria and gastrointestinal nematodes were found in 77% of the kids, 86% of the young goats and 71% of adult goats. The mean oocyst counts per gram of faeces in kids, young goats and adult goats were 9728, 1946, and 2667, respectively. The oocyst counts were not different significantly between age groups (P>0.05). The mean egg counts per gram of faeces in kids, young goats and adult goats were 1217, 1641 and 1092, respectively. The egg counts in kids were significantly lower than that in young goats (P<0.05). The intensity of Eimeria and gastrointestinal nematode infections between herds were significantly different (P<0.01).     

Corynebacterium Pseudotuberculosis Infection in Goats III.

Serological examinations for Gorynebacterium pseudotuberculosis in-fection were carried out in 14 known positive herds and in 1 herd that had beeni recently established through purchase of animals from different herds. Serum samples were collected sequentially on up to 4 occasions from animals during their first year of life. In most herds, these animals were examined clinically for superficial swellings once or twice during the same period. The adult goats in 8 infected herds were examined both clinically and serologically. Age distribution was similar in each of these herds. All serum samples were examined for antibodies against Gorynebac-terium pseudotuberculosis in both the bacterial agglutination test (BAT) and the hemolysis inhibition test (HIT). The proportion of kids which were seropositive in both tests de-creased to zero at 4 months of age. At the age of 11–12 months, the proportion of seropositive animals was about 50 % in the BAT and 30 % in the HIT. When examined when housed for the winter at the age of about 8 months (mean age), the prevalence of animals with superficial swellings was 7 %. At the age of about 1 year (mean age), 29 % of the examined animals had such lesions. In the recently established herd, only a few of the yearlings were seropositive. Titre values in BAT and HIT increased until 6 years of age. Anti-body titre values were significantly lower in yearlings than in older goats in both tests, P < 0.005. No significant difference in the propor-tion of goats with superficial swellings was seen at the different ages.     

Antibodies recognizing specific Mycobacterium avium subsp. paratuberculosis’s MAP3738c protein in type 1 diabetes mellitus children are associated with serum Th1 (CXCL10) chemokine

Recently Mycobacterium avium subsp. paratuberculosis (Map) was associated to type 1 diabetes mellitus (T1DM). In this study we investigated for Map presence in children affected by T1DM compared to healthy children. A pool of 212 sera from T1DM children at onset was compared to sera from 57 healthy children for humoral immune response towards the Map specific protein MAP3738c by ELISA. Serum concentrations of CXCL10 (pro-Th1) and CCL2 (pro-Th2) chemokines were also measured in both sera pool. Results showed that T1DM children had a stronger seropositivity towards MAP3738c protein compared to healthy children. Data highlighted also the correlation between serum activity of T1DM patients towards the specific protein of Map and the increase of CXCL10 concentration if compared to non-diabetic subjects. In conclusion, an immune response to Map in T1DM patients at onset was observed and this may indicate a role of the bacterium in triggering or precipitating the disease.     

Seroprevalence of Toxoplasma gondii infection in goats by the indirect haemagglutination, immunofluorescence and immunoenzymatic tests in the region of Uberlândia, Brazil

A comparative study of the indirect haemagglutination (IHA), immunofluorescence (IFAT) and immunoenzymatic (ELISA) tests was carried out to determine the prevalence of Toxoplasma gondii antibodies in goats. One hundred seventy-four serum samples were obtained from four goat herds from the region of Uberlandia, State of Minas Gerais. The distribution of the animals, according to their origin, was as follow: 71 from herd I; 39 from herd II; 37 from herd III; and 27 from herd IV. Serum samples were analyzed by IHA, IFAT and ELISA, considering the reactivity of the serum samples at dilution > or = 1:64 as cut off titer for the three tests. A global seroprevalence of 18.4% was observed, with significantly higher positivity rate in the herd II (66.7%) and older animals (> 36 months). A high and significant positive correlation was found between the titers obtained by the IHA versus IFAT, IHA versus ELISA, and ELISA versus IFAT. Therefore, it can be concluded that the three analyzed tests have shown to be highly concordant and appropriate for epidemiological surveys of Toxoplasma infection in goats. Although the seroprevalence of T. gondii infection in goats is relatively low in this region as compared to other regions of the country, adequate management might be useful and essential to control the infection in the goat herds.     

Detection of Mycobacterium avium subsp. paratuberculosis in goat and sheep flocks in Mexico

The slow growth of Mycobacterium avium subsp. paratuberculosis (Map) has hindered its investigation. No data concerning the presence of paratuberculosis in Mexico are currently available. The aim of this study was to identify M. avium subsp. paratuberculosis infection in sheep and goat flocks from several states in Mexico. Twenty-two animals, all manifesting clinical signs of paratuberculosis were obtained from six different Mexican states. Of these, 14 sheep and 8 goats were serologically diagnosed and multibacillary type lesions were identified. On the basis of mycobacteria concentration technique used for ileal mucosa, Map identification was confirmed by means of PCR and bacterial culture.In addition, samples were classified according to the abundance of acid-fast bacteria (AFB) identified in the intestinal mucosa and by the final concentrate of mycobacteria and the amount of DNA obtained. All correlations concerning the abundance of AFB in tissue, AFB recovered in the final concentrate and the amount of DNA obtained were recorded. These findings will allow to predict the amount of DNA which can be obtained by referring to the abundance of acid-fast organisms found in the tissue and, therefore, define the potential of using this method for the purpose of Map characterization. The results of this study indicate that paratuberculosis is widespread among goats and sheep in Mexico.     

Prevalence of mycoplasmas in external ear canal of goats: Influence of the sanitary status of the herd

A study was carried out in 15 French goat herds to assess the presence of mycoplasmas in the external ear canal of goats and to estimate the association between the ear isolates (highly pathogenic or non-pathogenic) and the status of the herds with respect to the mycoplasma infections. In each herd, 20 clinically normal goats were investigated once and mycoplasma isolates were identified by MF-dot tests. Influence of herd status on the proportion of animals carrying highly pathogenic species of mycoplasmas was evaluated by calculating the Odds Ratios. The present study gave first of all an estimate of the relative importance of mycoplasma carriage in healthy French dairy goat herds. The species isolated were MmmLC, Mcc, Ma, Mp (considered as highly pathogenic) and My and Mc (non-pathogenic species). Carriage often involve several strains and/or species in the same ear canal. The highly pathogenic species were significantly more frequent in the herds with prior history of mycoplasma infection. This asymptomatic carrier state makes the sanitary status of a herd ambiguous and represents a risk of subsequent propagation of the etiological agents, during close physical contact or by intervention of vectors (mites).     

Evaluation of an indigenous ELISA for diagnosis of Johne’s disease and its comparison with commercial kits

Country lacks sensitive and indigenous diagnostic kits for the screening of goats and sheep against Johne’s disease. Therefore an indigenous ELISA kit was developed using protoplasmic antigen from native Mycobacterium avium subspecies paratuberculosis ‘Bison Type’ strain of goat origin (Kit 1). In the present study, kit 1 and two commercial kits (kit 2 and 3) were evaluated with respect to ‘Gold Standard’ fecal culture in 71 animals (55 goats and 16 sheep). Kit 1 using indigenous antigen (protoplasmic antigen) was sensitive at very low concentration (0.1 μgm / well) as compared to purified commercial protoplasmic antigen (4 μgm / well) used in kit 2, in the Type 1 reactors (strong positive as positive). Screening of 71 animals by fecal culture detected 38.0% animals (goats-40.0%, sheep-31.2%) as positive (clinical shedders of bacilli) from these farm animals. Of the farm animals located at Central Institute for Research on Goats, herds of goat were endemic whereas, sheep flocks were comparatively resistant to Johne’s disease. The 29.5 and 61.9, 15.4 and 57.7 and 4.2 and 14.0% animals (goats and sheep) were in the category of sero-reactors type 1 and 2 of the ELISA kits 1, 2 and 3, respectively. In the type 1 sero-reactors, sensitivity and specificity of kit 1, 2 and 3 was 53.7 and 86.0, 17.8 and 86.0 and 3.5 and 94.7%, respectively. Indigenous ELISA test (kit 1) was significantly superior for the screening of goatherds and sheep flocks against JD as compared to commercial ELISA kits (Kit 2 and 3). In comparison to kit 2 and 3, kit 1 had highest sensitivity, comparable specificity and substantial to nearly perfect proportional agreement (Kappa Scores) with respect to ‘Gold standard’ fecal culture in goats and sheep. Disease being endemic in herds and flocks screened using ELISA kits, Type I sero-rectors had better correlation with fecal culture in comparison to Type II sero-reactors therefore, used for estimation of sero-prevalence. Newly developed Indigenous ELISA kit was simple, inexpensive, sensitive and reliable for screening of goats and sheep population against Johne’s disease. The study reports high prevalence of Johne’s disease in farm goatherds and sheep flocks, using sensitive tests (fecal culture and ELISA kit). Results of Type 1 reaction in kit 1 were optimally correlated with culture and were good for estimating the sero-prevalence. For controlling Johne’s disease in endemic herds initial removal of the animals in strong positive category (Tyep 1 reactors), may help to remove heavy shedders.     

Combined eradication strategy for CAE in a dairy goat farm in Japan

We conducted an eradication program from 2002 to 2006 against caprine arthritis-encephalitis (CAE) virus (CAEV) in an important farm that maintained goat breeds and had a high prevalence of CAEV infection. The program did not involve the slaughter and replacement of entire flocks, but rather the prevention of both vertical and horizontal transmission. The program consisted of (1) removal of kids immediately after birth, (2) segregation of each generation, and (3) culling of positive goats in periodical tests. All goats born before 2002 were regarded as infected and grouped into herd A. Kids born during the program were divided into several herds on the basis of CAEV infection risk, raised with calf milk replacer, and periodically tested by polymerase chain reaction (PCR) and the agar gel immunodiffusion (AGID) test. A total of 205 kids were produced from 137 parents in herd A, 92 of which were distinctly infected. Only 11 of the 205 kids were infected with CAEV and were culled. The remaining 194 kids and all other kids born from other herds were negative by PCR and AGID testing throughout the program. The milk yield of primiparous does was significantly increased after the eradication program. These findings indicate that the combine use of isolated and milk-deprived rearing and periodical detection testing are effective in establishing a CAEV-free flock from an infected flock.     

Evaluation of Two PCR Tests for Coxiella burnetii Detection in Dairy Cattle Farms Using Latent Class Analysis

Different tests performed on bulk tank milk samples (BTM) are available to determine the C. burnetii status of herds. However, these tests, which are based on the detection of either antibodies directed against C. burnetii (ELISA) or bacterial DNA (PCR), have limitations. A currently disease-free herd infected in the past may continue to test positive with ELISA due to the persistence of antibodies in animals that were infected and that subsequently cleared the infection. Infectious herds can also be misclassified using PCR because of the absence of bacteria in the BTM when the test is performed. Recently, PCR has been used for bacterial DNA detection in the farm environment, which constitutes the main reservoir of C. burnetii. The objectives of this study were to assess and compare the sensitivities and specificities of one commonly used PCR test in BTM (PCR BTM) and of a PCR applied to environmental samples (PCR DUST) in dairy cattle farms. BTM and dust samples were collected (using environmental swabs) in 95 herds. The evaluation of the performance of the 2 tests was conducted using latent class models accounting for within herd disease dynamics. Parameter estimation was carried out using MCMC, within a Bayesian framework. Two types of priors were used for the specificity of PCR DUST. A model with a uniform prior on 0–1 fitted the data better than a model with a uniform prior on 0.95–1. With the best model PCR DUST had a lower sensitivity than PCR BTM (0.75 versus 0.83) and a specificity of 0.72. The moderately low value for the specificity of PCR DUST suggests that the presence of bacteria on farm is not always associated with persistent infections and shedding of bacteria in milk.     

Epidemiological studies on caprine arthritis-encephalitis virus infection in Jordan

To investigate the seroprevalence of, and risk factors for, caprine arthritis-encephalitis virus (CAEV) infection in different breeds of goats in Jordan, sera from 1100 goats from three different geographical regions in Jordan were analyzed. Prevalence of antibodies to CAEV was determined using a competitive ELISA test. A semi-structured questionnaire was used to collect information on herd health and management. Questionnaire data were tested in a multivariable logistic regression model to elucidate risk factors associated with CAEV seropositivity. In addition, the incidence of CAEV antibodies was investigated in six goat herds located in the northern part of Jordan. Out of the 69 goat herds investigated, 16 (23.2%) had antibodies against CAEV. Individual goat true seroprevalence to CAEV was 8.9%. The highest CAEV seroprevalence was observed in goats older than 3-years and younger than 6-years of age. The seroprevalence of CAEV in goats was significantly higher (P < 0.05) in the northern part of Jordan than that in central or southern parts of Jordan. The multivariable logistic regression model identified large herd size (OD = 2.0; 95% CI: 1.1, 2.7), addition of new animals to the herd (OD = 1.3; 95% CI: 0.3, 1.6) and contact with other goat herds (OD = 1.1; 95% CI: 0.9, 2.0) as risk factors for CAEV seropositivity. The incidence of CAEV seropositivity in the six herds monitored in the northern part of Jordan ranged from 2.4 to 5.3%.     

Evaluation of Paracoccidioides brasiliensis Infection in Dairy Goats

Paracoccidioides brasiliensis is the etiological agent of paracoccidioidomycosis, a systemic mycosis that affects mainly rural workers in Brazil and other Latin American countries. The participation of domestic and wild animal species in the ecoepidemiology of paracoccidioidomycosis is not well understood. The objective of this study was to evaluate P. brasiliensis infection in dairy goats. The humoral immune response to the gp43 antigen, the main antigen used for paracoccidioidomycosis serodiagnosis and seroepidemiology, was evaluated in two goats immunized with inactivated P. brasiliensis yeast cells. Both animals produced antibodies against the P. brasiliensis gp43 antigen, detected by ELISA, 2 weeks after immunization. A total of 202 goat serum samples were analyzed by ELISA and the immunodiffusion test using P. brasiliensis gp43 and exoantigen as antigens. The seropositivity observed by ELISA was 26.2 % although no reactivity was detected by immunodiffusion. The animals over 18 months of age showed significantly higher positivity (40 %) than animals aged 6–18 months (14.8 %) and 0–6 months (2.6 %). Taking into account that cross-reactivity may occur with other pathogens, the serum samples were also analyzed by ELISA using Histoplasma capsulatum exoantigen as antigen and the positivity observed was 14.3 %. The low correlation (0.267) observed between reactivity to P. brasiliensis gp43 and H. capsulatum exoantigen suggests co-infection rather than cross-reactivity. This is the first report showing serological evidence of P. brasiliensis infection in goats and reinforces that domestic animals are useful epidemiological markers of paracoccidioidomycosis.     

Seroepidemiological study of Q fever in domestic ruminants in semi-extensive grazing systems

Background  

Q fever, a worldwide zoonotic disease caused by Coxiella burnetii, is endemic in northern Spain where it has been reported as responsible for large series of human pneumonia cases and domestic ruminants' reproductive disorders. To investigate pathogen exposure among domestic ruminants in semi-extensive grazing systems in northern Spain, a serosurvey was carried out in 1,379 sheep (42 flocks), 626 beef cattle (46 herds) and 115 goats (11 herds). Serum antibodies were analysed by ELISA and positive samples were retested by Complement Fixation test (CFT) to detect recent infections.     

Typing of Mycobacterium avium Subspecies paratuberculosis Isolates from Newfoundland Using Fragment Analysis

Short Sequence Repeat (SSR) typing of Mycobacterium avium subspecies paratuberculosis (Map) isolates is one of the most commonly used method for genotyping this pathogen. Currently used techniques have challenges in analyzing mononucleotide repeats >15 bp, which include some of the Map SSRs. Fragment analysis is a relatively simple technique, which can accurately measure the size of DNA fragments and can be used to calculate the repeat length of the target SSR loci. In the present study, fragment analysis was used to analyze 4 Map SSR loci known to provide sufficient discriminatory power to determine the relationship between Map isolates. Eighty-five Map isolates from 18 animals from the island of Newfoundland were successfully genotyped using fragment analysis. To the best of our knowledge, this is the first report on Map SSR diversity from Newfoundland dairy farms. Previously unreported Map SSR-types or combinations were also identified during the course of the described work. In addition, multiple Map SSR-types were isolated from a single animal in many cases, which is not a common finding.     

An agar--gel immunodiffusion test for detection of Brucella antibodies in human serum.

A comparison was made of results obtained with a Brucella agar--gel immunodiffusion (AGID) test and the standard tube-agglutination test on 612 human sera. Agreement between the tests was 97% when the titer was 1:160 or higher. Of 448 sera that showed no agglutination titer, 447 were negative with the AGID test. Results of the AGID test were also compared to those obtained with the 2-mercaptoethanol (2-ME) agglutination test on 148 sera that demonstrated a standard tube-agglutination titer of 1:20 or higher. All sera with a 2-ME-agglutination titer of 1:40 or higher were positive with the AGID test. Of 123 sera that showed no 2-ME-agglutination titer, 21 were positive with the AGID test. Two of these 21 sera were obtained from patients with bacteriologically proven brucellosis, and eight were from abattoir employees with suspected but not bacteriologically proven brucellosis.