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Distribution of IS1311 and IS1245 in Mycobacterium avium subspecies revisited.

We demonstrated that IS1245 is not present in Mycobacterium avium subsp. paratuberculosis by restriction fragment length polymorphism and that the designated three-banded bird pattern of IS1245 in M. avium subsp. avium consists of one copy of IS1245 and two copies of IS1311. Cross hybridization between the two elements can be avoided by using more specific probes.

Animals↗

Effect of intestinal resection on serum antibodies to the mycobacterial 45/48 kilodalton doublet antigen in Crohn's disease.

Interest in the role of mycobacterial infection in Crohn's disease has been revived by the cultural detection of Mycobacterium paratuberculosis in patients with Crohn's disease. This hypothesis was examined serologically using assays with high specificity for Crohn's disease. The effect of intestinal resection on serum antibodies specific for Crohn's disease was investigated with an immunoblot assay and an enzyme linked immunosorbent assay using the 45/48 kilodalton doublet antigen of Mycobacterium tuberculosis. Antibodies were detected in 64.7% of patients with Crohn's disease (n = 17), 10% of patients with ulcerative colitis (n = 10), 5% of patients with carcinoma of the colon (n = 20), and none of 10 healthy subjects with the immunoblot assay. Statistical comparison of the Crohn's disease patients with each control group resulted in p = 0.0000236. Immunoglobulin G was essentially unchanged 75 days (mean) after surgery. After more than 180 days, however, the antibody response was reduced in all of five patients studied, and was no longer demonstrable in two of them (40%). Simultaneously, the Crohn's disease activity index (CDAI) decreased. Both the high specificity of this assay for Crohn's disease and the diminished antibody response after intestinal resection in parallel with decreased CDAI support a mycobacterial aetiology of Crohn's disease.

Adolescent↗

Controlled trial of antituberculous chemotherapy in Crohn's disease: a five year follow up study.

BACKGROUND: It has been suggested that Mycobacterium paratuberculosis is the cause of Crohn's disease. In a previous report the immediate effect of two years treatment with antituberculous chemotherapy showed no clinical benefit. AIMS: To assess both the immediate and longer term effect of treatment on the disease. METHODS: Patients were followed for five years from their date of entry to the study. One hundred and thirty patients entered the initial study, and of these 111 (81%) were followed regularly. RESULTS: Overall, there was no evidence of consistent benefit or disadvantage from antituberculous chemotherapy in any of the assessments made, including the number of acute relapses, surgical episodes, hospital admissions, disease activity, blood tests, or medication required for Crohn's disease during the follow up period. CONCLUSION: The absence of any benefit at the end of the initial two year trial period, and during the three year subsequent follow up, fails to support the hypothesis that mycobacteria play an important part in the pathogenesis of Crohn's disease, or that antituberculous chemotherapy may be of benefit.

Antitubercular Agents↗

Induction of humoral immunity in response to immunization with recombinant Mycobacterium bovis BCG expressing the S1 subunit of Bordetella pertussis toxin.

Two recombinant Mycobacterium bovis BCG (rBCG) vaccine strains were developed for the expression of cytoplasmically located S1 subunit of pertussis toxin, with expression driven by the hsp60 promoter of M. bovis (rBCG/pPB10) or the pAN promoter of Mycobacterium paratuberculosis (rBCG/pPB12). Both strains showed stable expression of equivalent levels of recombinant S1 in vitro and induced long-term (up to 8 months) humoral immune responses in BALB/c mice, although these responses differed quantitatively and qualitatively. Specifically, rBCG/pPB12 induced markedly higher levels of IgG1 than did rBCG/pPB10, and mice immunized with the former strain developed specific long-term memory to S1, as indicated by the production of high levels of S1-specific IgG in response to a sublethal challenge with pertussis toxin 15 months after initial immunization. When considered in combination with previous studies, our data encourage further evaluation of rBCG as a potential means of developing a low-cost whooping cough vaccine based on defined antigens.

Animals↗

The seroprevalence of Johne's disease in Georgia beef and dairy cull cattle.

Beef and dairy cattle serum samples, collected during 2000 at sale barns throughout Georgia, were obtained from the Georgia State Brucellosis Laboratory and were used to conduct a retrospective epidemiological study. Statistical samplings of 5,307 sera, from over 200,000 sera, were tested for antibodies to Mycobacterium avium ssp. paratuberculosis, (Johne's disease) using a commercial enzyme-linked immunosorbent assay test kit. An overall period seroprevalence in all classes of cattle tested was 4.73%. The period seroprevalence in dairy cattle was 9.58%, in beef cattle it was 3.95%, and in cattle of unknown breed it was 4.72%. It was concluded that the seroprevalence of Johne's disease in cull beef and dairy cattle in Georgia is economically significant.

Animals↗

Fluid dairy product quality and safety: looking to the future.

The fiercely competitive nature of the US beverage industry will drive the fluid milk sector of the dairy industry to improve product quality and shelf life to enable dairy beverages to compete with innovative new introductions as well as with currently popular shelf-stable products. The recent substantial growth in the volume of flavored milk sales specifically suggests that attention is needed to improve these products. Further, increasing public awareness and regulatory attention directed toward food safety issues highlight the need for the dairy industry to proactively address and eliminate emerging food safety issues that may negatively impact the image of dairy products. Shelf life and sensory profiles of high temperature short time pasteurized fluid milk products are presented, illustrating the need for greater attention to controlling contaminating microorganisms in processed fluid milk products. Bacterial spoilage patterns of flavored versus unflavored milks are compared, and suggestions are presented for extending flavored product shelf lives. Strategies currently applied to extend shelf life are reviewed. Food safety issues facing the dairy industry are presented within the context of an overview of foodborne illnesses in the United States. The pressing need to determine thermal resistance characteristics of Mycobacterium paratuberculosis is described.

Animals↗

Effects of supplemental energy on metabolic and immune measurements in periparturient dairy cows with Johne's disease.

The present study was designed to evaluate whether feeding supplemental energy would improve the metabolic profile and alleviate some of the immunosuppression typically noted during the periparturient period in dairy cows with Johne's disease. Twelve dairy cows naturally infected with Mycobacterium paratuberculosis were fitted with rumen cannulas in late gestation and assigned to treatment groups: control, n = 6; or stuffed, n = 6. Cows in the control group were allowed to consume feed ad libitum. Cows assigned to the stuffed treatment group were also fed ad libitum but received additional total mixed rations by manually stuffing their rumens with refused feed to maintain dry matter intake of 2% body weight per day before calving and 2.5% body weight per day after calving. Serum nonesterified fatty acid levels were significantly decreased in stuffed cows compared with control cows, indicating that stuffing to maintain dry matter intake improved the energy balance in the cows. In addition, periparturient serum calcium and magnesium concentrations were significantly higher in stuffed cows. Stuffing modulated cell-mediated immunity by reducing lymphocyte proliferative responses to T-cell mitogens during early lactation. Stuffing resulted in an increase in the secretion of in vitro immunoglobulin by peripheral blood mononuclear cells after parturition when compared to control cows. These data demonstrate that energy balance is improved by providing additional energy in this manner and suggest energy supplementation can improve some aspects of immune function during the periparturient period.

Animal Feed↗

Disinfection of dairy and animal farm wastewater with radiofrequency power.

Radiofrequency (RF) power was investigated as a new, physical (nonchemical), thermal process to disinfect wastewater from dairy and animal facilities. Samples (n = 38) from 8 dairy, 2 calf, and 3 swine facilities in California were collected over a 3-yr period and characterized for their dielectric properties, chemical composition, and suitability for thermal processing using RF power. To establish efficacy for disinfection, selected samples were inoculated with high levels (10(6) to 10(9) cfu/mL) of bacterial pathogens such as Salmonella sp., Escherichia coli O157:H7, and Mycobacterium avium ssp. paratuberculosis and processed with an RF prototype system. The capabilities of RF power as a method for thermal disinfection of wastewater were demonstrated when bacteria pathogens were completely and rapidly (<1 min) inactivated when temperatures of 60 to 65 degrees C were achieved. Furthermore, RF technology can be used for large-scale, batch or continuous and portable applications, allowing significant improvements in energy-use efficiencies compared with conventional thermal (surface heating) technologies. Therefore, RF power has potential as an alternative to disinfect dairy/animal farm wastewater before recycling.

Agriculture↗

Survey of free-ranging elk from Wyoming and Montana for selected pathogens.

From December 1991 through January 1995, a disease survey was conducted on herds of free-ranging, hunter-killed elk (Cervus elaphus nelsoni) from three areas in proximity to Yellowstone National Park (YNP), Wyoming (USA), after tuberculosis caused by Mycobacterium bovis was discovered in a captive herd of elk in the area. Complete or partial sets of specimens from 289 elk collected between December 1991 and January 1993 were examined histologically; no mycobacterial lesions were observed. Lesions of tuberculosis were not detected in tonsils or lymph nodes of the head from an additional 99 hunter-killed, adult elk from one area (area 2) collected in January 1995. Neither M. bovis nor M. paratuberculosis were isolated from any of the specimens cultured. Antibodies to Brucella abortus were detected in serum samples from 0%, 1%, and 1% of elk from three areas sampled (areas 1, 2 and 3), respectively. Brucella abortus biovar 1 was isolated from multiple tissues from one seropositive animal from area 3. Larvae with morphology consistent with Dictyocaulus sp. were found in 12%, 14%, and 0% of fecal specimens tested from areas 1, 2, and 3, respectively. Pasteurella multocida and Actinomyces pyogenes were isolated from a lung with purulent bronchopneumonia and abscesses.

Animals↗

Rapid detection of single cell bacteria as a novel approach in food microbiology.

Solid-phase cytometry (SPC) is a novel technique that allows rapid detection of bacteria at the single cell level, without the need for a growth phase. After filtration of the sample, the retained microorganisms are fluorescently labeled on the membrane filter and automatically counted by a laser scanning device. Each fluorescent spot can be visually inspected with an epifluorescence microscope connected to the ChemScan by a computer-driven moving stage. Depending on the fluorogenic labels used, information on the identity and the physiological status of the microorganisms can be obtained within a few hours. Although SPC was originally recommended for the determination of the total viable microbial count in water and other liquid samples, it may also be a promising technique for the detection and enumeration of bacteria in food samples, provided they can be isolated from the unfilterable matrix. The short detection time inherent in this approach is a considerable advantage over conventional plate counting, especially for slow-growing microorganisms. The basic principles of SPC are discussed as well as its potential for the detection of Mycobacterium paratuberculosis, a model example of a slow-growing bacterium in milk.

Animals↗

Integrating theories of the etiology of Crohn's disease. On the etiology of Crohn's disease: questioning the hypotheses.

The most prominent theory describes the Crohn's Syndrome as a dysregulated, inappropriate immune response to otherwise innocuous bowel flora in a genetically susceptible host. The autoimmune theory assumes that a specific infectious agent does not exist. Data from mouse models, impairment of the mucosal epithelial barrier and the influence of gut flora are used to support the autoimmune theory. Critics claim that the dysregulated immune responses are not the primary disorder but secondary to an underlying infection. Two other theories are also consistent with the same data. The immunodeficiency theory hypothesizes that defects in innate immunity leading to compensatory immune processes underlie the Crohn's phenotype and suggests that therapy should stimulate immunity rather than suppress it. The mycobacterial theory proposes that Mycobacterium avium subspecies paratuberculosis is one of the causes of the Crohn's Disease syndrome. Mycobacterial molecules dysregulate immune signaling pathways as part of the organisms'evolved survival strategy. If MAP were to initiate the dysregulated immune response then the necessity to hypothesize that commensal gut flora provide the antigenic stimulus would be moot. Commensal bacteria would be relegated to a secondary role of modifying the immune response rather than occupying the central role of providing the initiating antigens. Critics claim that MAP is merely a secondary invader. The three theories differ primarily by emphasizing different aspects of the same overall process.

Animals↗

Insertion elements in mycobacteria.

We have isolated and characterised a repetitive element from the genome of Mycobacterium paratuberculosis. This repetitive element shows many features characteristic of a bacterial insertion element.

Cloning, Molecular↗

[Pathogenesis, clinical aspects and diagnosis of the most important enteric bacterial diseases in animals].

There are adhesive and invasive species among enteropathogenic bacteria for animals. Most frequently isolated adhesive bacteria are enterotoxigenic E. coli (ETEC). Salmonella, T. hyodysenteriae, M. paratuberculosis, Y. enterocolitica, Cl. perfringens and C. jejuni bacteria represent the most important invasive germs. The main clinical finding connected with bacterial enteritis is diarrhoea resp. dysentery. Evaluation of feces consistency, time of appearance and accompanying symptoms (e.g. fever, colic) allow a clinical suspective diagnosis, however a definitive diagnosis must be based on the demonstration of the causative agent. In addition to known cultural, microscopic, biochemical and serological diagnostic methods, the demonstration of virulence factors (enterotoxin formation, adhesive pili) plays an important role in diagnosis of enteropathogenic bacteria.

Animals↗

The use of a single complement fixation test technique in bovine brucellosis, Johne's disease, dourine, equine piroplasmosis and Q fever serology.

The same techniques may be used in the complement fixation test (CFT) for the serological diagnosis of bovine brucellosis, Johne's disease (paratuberculosis), dourine, equine piroplasmosis and Q fever (caused by Coxiella burnetii). The reproducibility of results is excellent, falling for the most part within the twofold range and never exceeding the fourfold range. Agreement with other laboratories is excellent (i.e. within twofold) in the case of brucellosis and equine piroplasmosis antibody titres. A good correlation between the occurrence of the disease and serological reactions is found on circumstantial evidence in the cases of dourine, Johne's disease and Q fever. A standard unitage system is used to report the antibody titres found in all the tests. To simplify laboratory protocols, laboratories required to employ the CFT for the diagnosis of these diseases are advised to use a single proven technique in all the tests. Problems experienced with transient false-positive Johne's disease antibody titres in cattle following on tuberculin (bovine and avian) testing make it advisable to take specimens for the Johne's disease test prior to performing the tuberculin tests.

Animals↗

Johne's disease and the practitioner.

Mycobacterium paratuberculosis infections and the disease states resulting in cattle are defined as spectral in nature. There is a spectrum of immunological response which complicates tests based on this. It is recommended that initial diagnosis of suspicious clinical cases be confirmed by direct fecal smears and a serological test such as immunodiffusion. Herd evaluation requires the application of fecal cultures plus immunodiffusion. Control requires rigid management hygiene measures, surveillance of the herd status and post mortem confirmation of salvaged animals.

Animals↗

Idiopathic eosinophilic enteritis in four cattle.

Four cattle had chronic diarrhea and had lost weight but were not anorectic or dehydrated. A diagnosis of eosinophillic enteritis was made on the basis of a combination of clinical and histopathologic findings and exclusion of other diseases. Eosinophilia and hypoalbuminemia were not detected in CBC analyses. Gross and histologic examination of the small intestine revealed edema, diffuse eosinophilic inflammation (> 20 eosinophils/HPF [400 x]), and regional lymphadenopathy. A causative agent was not identified in any of the cattle. Treatment consisting of a 1-month course of systemically administered corticosteroids was attempted in 2 cattle and was apparently successful in both. Diagnostic criteria for eosinophilic enteritis have not been established for cattle and, therefore, the diagnosis was subjective. This disease resembled paratuberculosis (Johne's disease) in clinical appearance but was distinguished on the basis of a lack of a contagious etiologic agent and differences in histologic appearance.

Adrenal Cortex Hormones↗

Long-term sequelae to foodborne disease.

Most of the concern about foodborne disease has been focused on the immediate effects of acute infection. Recent information has shown that many of these foodborne infections also have long-term sequelae with serious health effects and a significant economic impact. To increase the awareness of animal health professionals to these sequelae, the authors discuss two groups of sequelae which are strongly associated with preceding infection (reactive arthritides, including Reiter's syndrome, and the Guillain-Barré syndrome) as well as the possible association between Crohn's disease and Mycobacterium paratuberculosis. The discussion includes a description of the disease syndromes along with epidemiological and economic information. More reliable epidemiologial and economic data on chronic sequelae to foodborne disease will be needed for future evaluation of the cost-effectiveness of mitigation strategies to reduce the occurrence of foodborne pathogens.

Animals↗