Yersinia pseudotuberculosis in an alpaca.

A 6-year-old female huacaya alpaca was referred to the clinic for evaluation with a 1-month history of rapid weight loss, inappetence, lethargy, and severe leukocytosis refractory to medical management. Physical examination revealed a body condition score of 1 out of 5 and a large, firm structure palpable in the right caudoventral abdomen. Abdominal ultrasonographic examination revealed 3 masses with hyperechoic, swirling centers. The largest mass measured 15 cm in diameter with a 2-centimeter capsule, and extended from right of midline into the left inguinal region. Transrectal ultrasonography identified a small uterus and clear delineation between the abdominal masses. Complete blood (cell) count findings were consistent with marked systemic inflammation. Based on initial examination and laboratory findings, exploratory laparotomy was elected. Multiple mesenteric masses strongly adhered to the jejunum were observed within the abdomen. Due to the inoperable conditions and the poor long-term prognosis, the alpaca was euthanized under general anesthesia. Bacterial culture of fluid aspirated from the largest mass revealed Yersinia pseudotuberculosis. Key clinical message: Clinical progression and attempted treatment of Yersinia pseudotuberculosis in camelids have not been previously described and the bacterium should be considered as a differential diagnosis for abscessation and persistent leukocytosis. Yersinia pseudotuberculosis is also considered a zoonotic agent and proper precautions should be taken when handling cases of abdominal abscessation.

Yersinia pseudotuberculosis chez un alpaga. Une alpaga huacaya femelle de 6 ans a été référée à la clinique pour évaluation avec des antécédents d'un mois de perte de poids rapide, d'inappétence, de léthargie et de leucocytose sévère réfractaire à la prise en charge médicale. L'examen physique a révélé un score d'état corporel de 1 sur 5 et une structure large et ferme palpable au niveau de l'abdomen caudoventral droit. L'examen échographique abdominal a révélé 3 masses à centres hyperéchogènes et tourbillonnants. La plus grande masse mesurait 15 cm de diamètre avec une capsule de 2 centimètres et s'étendait de la droite de la ligne médiane jusqu'à la région inguinale gauche. L'échographie transrectale a identifié un petit utérus et une délimitation claire entre les masses abdominales. Les résultats de la numération globulaire (cellulaire) sanguine complète étaient compatibles avec une inflammation systémique marquée. Sur la base de l'examen initial et des résultats de laboratoire, une laparotomie exploratoire a été choisie. De multiples masses mésentériques fortement adhérées au jéjunum ont été observées dans l'abdomen. En raison des conditions inopérables et du mauvais pronostic à long terme, l'alpaga a été euthanasié sous anesthésie générale. La culture bactérienne du liquide aspiré de la plus grande masse a révélé Y. pseudotuberculosis.Message clinique clé :La progression clinique et les tentatives de traitement de Y. pseudotuberculosis chez les camélidés n'ont pas été décrites auparavant et la bactérie doit être considérée comme un diagnostic différentiel d'abcès et de leucocytose persistante. Yersinia pseudotuberculosis est également considérée comme un agent zoonotique et des précautions appropriées doivent être prises lors de la manipulation des cas d'abcès abdominal.(Traduit par Dr Serge Messier).

Interactive effects of dexamethasone and opsonized Mycoplasma bovis on bovine neutrophil function in vitro.

Previously we had reported that exposure to high levels of glucocorticoids, and to unopsonized Mycoplasma bovis, has a negative interactive effect on bovine neutrophil function in vitro, and this interactive effect was a function of M. bovis strain differences. Here we hypothesized that in vitro treatment of bovine neutrophils by glucocorticoid would impair phagocytosis of opsonized M. bovis compared to non-treated neutrophils and such impairment would be a function of M. bovis strain differences. Neutrophils isolated from 20 mid-lactation cows were treated with immunosuppressive dose of 5 × 10-4 M dexamethasone or placebo and incubated with one of four opsonized M. bovis strains that had been isolated from bovine origin. After incubation neutrophil function measured included: percentage reduction in log10 of M. bovis CFU/ml, percentage of phagocytizing neutrophils, phagocytized M. bovis per neutrophil, and killed M. bovis per neutrophil. Least square means of all neutrophil groups were contrasted using linear mixed-effects models. Effects due to strain, treatment, and their interaction on neutrophil function measured by the number of phagocytized M. bovis per neutrophil and number of killed M. bovis per neutrophil were different (P < 0.05). However, no significant strain by treatment interaction effect on percentage reduction in log10 of M. bovis CFU/ml was found. Neither a strain nor a strain by treatment interaction was found to affect the percentage phagocytizing neutrophils. These findings might explain in part the association of stressful events with subsequent outbreaks of Mycoplasma bovis associated bovine diseases.

Bluetongue disease and seroprevalence in South American camelids from the northwestern region of the United States.

In late summer/early fall of 2013, 2 South American camelids from central Washington were diagnosed with fatal bluetongue viral disease, an event which is rarely reported. A 9-year-old intact male llama (Lama glama), with a 1-day history of anorexia, recumbency, and dyspnea before death. Abundant foam discharged from the mouth and nostrils, and the lungs were severely edematous on postmortem examination. Histologically, there was abundant intra-alveolar edema with fibrin. Hemorrhage and edema disrupted several other organs. Bluetongue viral RNA was detected by reverse transcription polymerase chain reaction (RT-PCR), and serotype 11 was identified by sequencing a segment of the VP2 outer capsid gene. Approximately 1 month later, at a site 150 miles north of the index case, a 2-year-old female alpaca with similar, acutely progressive clinical signs was reported. A postmortem examination was performed, and histologic lesions from the alpaca were similar to those of the llama, and again serotype 11 was detected by PCR. The occurrence of bluetongue viral infection and disease is described in the context of seasonal Bluetongue virus activity within the northwestern United States and southwestern Canada.

Effects of dexamethasone and Mycoplasma bovis on bovine neutrophil function in vitro.

It is well established that exposure either to elevated levels of glucocorticoids, or to Mycoplasma bovis (M. bovis), has a negative effect on bovine neutrophil function. The objective of this research was to determine whether in vitro treatment of bovine neutrophils by M. bovis strains (n=4) and glucocorticoids would additively impair phagocyte function. Twenty, healthy, dairy cows were enrolled. Whole blood was collected from all cows for neutrophil isolation. Phagocytosis and the generation of superoxide anion (O2(-)) were tested in vitro by incubation of neutrophils with FITC labeled Escherichia coli (E. coli) and cytochrome c after treatment. Treatments included: NM1-4D (neutrophils treated with dexamethasone and exposed to one of the four M. bovis strains); NM1-4 (neutrophils exposed to one of the four M. bovis strains only); ND (neutrophils treated with dexamethasone only); and N (non-treated control neutrophils). The overall percentages of neutrophils phagocytizing E. coli were: 32%, 51%, 37%, and 53% ± 5.25% for treatments NM1-4D, NM1-4, ND, and N, respectively. The overall statistically transformed means of phagocytized E. coli per neutrophil were: 1.37, 1.72, 1.33, and 1.67 ± 0.057 for treatments NM1-4D, NM1-4, ND, and N, respectively. The overall statistically transformed means of neutrophil O2(-) production were: 8.60, 11.91, 9.01, and 12.21 ± 0.21 nmol/10(6) for treatments NM1-4D, NM1-4, ND, and N, respectively. Exposure of neutrophils to M. bovis plus dexamethasone had an additive effect on generation of reactive oxygen species (p=0.0057), but not on the percentage of neutrophils phagocytizing E. coli (p=0.0817) or number of E. coli phagocytized per neutrophil (p=0.2946). Only one of the four M. bovis strains had a negative effect on neutrophil phagocytic function. Dexamethasone treatment consistently decreased neutrophil function as indicated by decreased percentage of neutrophils phagocytizing E. coli, decreased number of E. coli phagocytized per neutrophil, and decreased neutrophil O2(-) production, compared to controls (p<0.0001). Results suggested a synergistic effect of in vitro incubation of glucocorticoids and M. bovis on reduction of bovine neutrophil function as measured by generation of reactive oxygen species. These findings may explain in part the interaction between stressful events and outbreak of Mycoplasma bovis associated bovine disease.

Deletion of relA abrogates the capacity of Mycobacterium avium paratuberculosis to establish an infection in calves.

Previous comparative studies in goats revealed deletion of relA but not pknG abrogates the capacity of Mycobacterium avium subsp. paratuberculosis (Map) to establish a persistent infection. The immune response elicited by the mutant cleared infection. The objective of the present study was to extend the studies in calves and compare the proliferative response elicited by the relA deletion mutant (ΔrelA) and Map using flow cytometry and quantitative reverse transcription real-time PCR (qRT-PCR). Six 3-day-old calves were divided into two groups. Three were vaccinated with ΔrelA and 3 inoculated with wild type Map. The calves were challenged with Map 1 month later and necropsied 3 months post challenge. Three untreated calves were used as uninfected controls. Examination of tissues revealed the ΔrelA mutant was immune eliminated. Bacterial load of Map was significantly reduced in the calves vaccinated with ΔrelA and challenged with Map in comparison with calves inoculated and challenged with Map. A vigorous CD4 memory T cell response was detected at necropsy in PBMC from both infected groups. CD8 positive NK cells proliferated in the presence and absence of antigen stimulation in both treated groups but not in the uninfected group. IFN-γ, IL17, and IL22 gene expression were up-regulated with an associated increase in their transcription factors, Tbet and RORC, in both treated groups. TGF-ß, IL-10, and FoxP3 were not up-regulated, indicating no activation of regulatory T cells. The findings show that the immune response to ΔrelA is clearly different than the response to Map. Understanding the immunological basis for this difference should facilitate development of a vaccine that elicits sterile immunity.

Safety and Immunogenicity of a Mycoplasma ovipneumoniae bacterin for domestic sheep (Ovis aries).

BACKGROUND: Mortality from epizootic pneumonia is hindering re-establishment of bighorn sheep populations in western North America. Mycoplasma ovipneumoniae, a primary agent of this disease, is frequently carried asymptomatically by the domestic sheep and goats that constitute the reservoir of this agent for transmission to bighorn sheep. Our long-term objective is to reduce the risk of M. ovipneumoniae infection of bighorn sheep; one approach to this objective is to control the pathogen in its reservoir hosts. METHODS: The safety and immunogenicity of M. ovipneumoniae for domestic sheep was evaluated in three experimental immunization protocols: 1) live M. ovipneumoniae (50 ug protein); 2) killed M. ovipneumoniae (50 ug whole cell protein) in oil adjuvant; and 3) killed M. ovipneumoniae (250 ug whole cell protein) in oil adjuvant. Immunogenicity was assessed by two serum antibody measures: competitive enzyme-linked immunosorbent assay (cELISA) (experiments 1-3) and serum growth inhibition (Experiment 3). Passive immunogenicity was also assessed in the third experiment using the same assays applied to blood samples obtained from the lambs of immunized ewes. RESULTS AND CONCLUSIONS: Adverse reactions to immunization were generally minor, but local reactions were regularly observed at immunization sites with bacterins in oil adjuvants. No evidence of M. ovipneumoniae specific antibody responses were observed in the first or second experiments and no resistance to colonization was observed in the first experiment. However, the ewes in the third experiment developed strong cELISA serum antibody responses and significant serum M. ovipneumoniae inhibition activity, and these responses were passively transferred to their lambs. The results of these trials indicate that immunization with relatively large antigenic mass combined with an adjuvant is capable of inducing strong active antibody responses in ewes and passively immunizing lambs.

Computed tomography findings in a 5-year-old Australian Cashmere goat (Capra hircus) suffering leukoencephalomyelitis due to caprine arthritis encephalitis virus.

Computed tomography was used to aid in the antemortem diagnosis of leukoencephalomyelitis in a goat infected by caprine arthritis encephalitis virus (CAEV). Imaging results were corroborated by histologic examination. This report discusses various methods of imaging the nervous system and their potential for use in the antemortem diagnosis of CAEV neurologic changes.

Résultats d'une tomodensitométrie chez une chèvre cachemire de l'Australie(Capra hircus)âgée de 5 ans souffrant d'une lymphoencéphalomyélite en raison du virus de l'arthrite caprine. La tomodensitométrie a été utilisée pour faciliter le diagnostic antemortem de la lymphoencéphalomyélite chez une chèvre infectée par le virus de l'arthrite-encéphalite caprine (VAE). Les résultats de l'imagerie ont été corroborés par l'examen histologique. Ce rapport discute les diverses méthodes d'imagerie du système nerveux et leur utilisation potentielle pour le diagnostic antemortem des changements neurologiques du VAE.(Traduit par Isabelle Vallières).

Traumatic gastroperitonitis (hardware disease) in an alpaca (Vicugna pacos).

A 5-yr-old, intact male Suri alpaca (Vicugna pacos) presented with acute onset of anorexia, depression, and reluctance to stand. Exploratory laparotomy revealed diffuse peritonitis resulting from penetration of a 5-cm wire through the second gastric compartment (C2). Due to the severity of the alpaca's condition and its deteriorating nature, euthanasia at the time of surgery was elected. This is the first published case of a condition commonly observed in cattle (traumatic gastroperitonitis, "Hardware disease") to be reported in an alpaca.

A multivalent Mannheimia-Bibersteinia vaccine protects bighorn sheep against Mannheimia haemolytica challenge.

Bighorn sheep (BHS) are more susceptible than domestic sheep (DS) to Mannheimia haemolytica pneumonia. Although both species carry M. haemolytica as a commensal bacterium in the nasopharynx, DS carry mostly leukotoxin (Lkt)-positive strains while BHS carry Lkt-negative strains. Consequently, antibodies to surface antigens and Lkt are present at much higher titers in DS than in BHS. The objective of this study was to determine whether repeated immunization of BHS with multivalent Mannheimia-Bibersteinia vaccine will protect them upon M. haemolytica challenge. Four BHS were vaccinated with a culture supernatant vaccine prepared from M. haemolytica serotypes A1 and A2 and Bibersteinia trehalosi serotype T10 on days 0, 21, 35, 49, and 77. Four other BHS were used as nonvaccinated controls. On the day of challenge, 12 days after the last immunization, the mean serum titers of Lkt-neutralizing antibodies and antibodies to surface antigens against M. haemolytica were 1:160 and 1:4,000, respectively. Following intranasal challenge with M. haemolytica A2 (1 × 10(5) CFU), all four control BHS died within 48 h. Necropsy revealed acute fibrinonecrotic pneumonia characteristic of M. haemolytica infection. None of the vaccinated BHS died during the 8 weeks postchallenge observation period. Radiography at 3 weeks postchallenge revealed no lung lesions in two vaccinated BHS and mild lesions in the other two, which resolved by 8 weeks postchallenge. These results indicate that if BHS can be induced to develop high titers of Lkt-neutralizing antibodies and antibodies to surface antigens, they are likely to survive M. haemolytica challenge which is likely to reduce the BHS population decline due to pneumonia.

The effects of exposure of susceptible alpacas to alpacas persistently infected with bovine viral diarrhea virus.

Reports of bovine viral diarrhea virus (BVDV) infections in alpacas have been increasing in recent years but much is still unknown about the mechanisms of disease in this species. This report characterizes the transmission of BVDV from persistently infected (PI) alpacas to BVDV naïve alpacas, documents shedding patterns, and characterizes the disease effects in both PI and transiently infected alpacas. Two PI alpacas shed BVDV Type 1b virus in most body fluids, and commonly available diagnostic tests verified their status. Bovine viral diarrhea virus Type 1b transient infections produced only mild signs of disease in BVDV naïve alpacas. Viremia was detected in whole blood, but viral shedding during the acute phase was not detected and antibody appeared to be protective upon re-exposure to the virus.

Experimental infection of a bovine model with human isolates of Mycobacterium avium subsp. paratuberculosis.

Mycobacterium avium subsp. paratuberculosis (Map), the etiologic agent of Johne's disease (JD) in ruminants, has been implicated in the pathogenesis of Crohn's disease (CD) in humans. We developed a bovine ileal cannulation model to facilitate comparison of the immune response to Map and the mechanisms of pathogenesis in cattle and humans. Initial studies showed a T cannula could be maintained for up to a year in calves without inducing inflammation or adversely affecting intestinal function. Map introduced through the cannula established a persistent low level of infection without inflammation. Infection elicited an immune response to Map antigens detectable by flow cytometry. Further studies now show the cannulation model can be used with cows during the later stage of infection, affording access to the target tissue at all stages of infection. The studies also revealed no difference in infectivity or immunogenicity of isolates of Map obtained from cattle or humans with CD. Comparison of the immune response to Map during the early and late stages of infection using PCR, flow cytometry and QRT-PCR, showed the immune response early in the disease process is dominated by CD4 T cells. A CD8 response is delayed but comparable at later stages of infection. Genes for pro-inflammatory cytokines IFN-γ and the recently identified genes encoding IL-17 and IL-22 are up regulated in infected animals. These findings reveal that both human and bovine isolates of Map can establish infection and induce similar immune responses in a bovine model. They also reveal the cytokine responses elicited in cattle are similar to those implicated in CD pathogenesis.

Diaphragmatic hernia in a two-year-old alpaca (Vicugna pacos).

A 2-yr-old male intact alpaca (Vicugna pacos) was admitted for a 4-day history of anorexia and colic. Five months prior, the alpaca had undergone surgical removal of a duodenal trichophytobezoar and had recovered uneventfully. The alpaca died under anesthesia, and diaphragmatic herniation of the third gastric compartment (C3) was diagnosed at necropsy. A defect was identified in the left dorsal hemidiaphragm accompanied by herniation of 80% of C3 and the aboral portion of the second gastric compartment into the pericardial sac. The smooth margins and dorsal location of the diaphragmatic defect suggested a congenital origin. Diaphragmatic herniation is uncommon in camelids, and only one other case has been reported. Due to the dorsal location of the diaphragmatic defect in this animal, positioning during the previous surgery may have initiated a partial entrapment of gastric compartments, leading to a more complete incarceration between when the animal was discharged and presented again.

Evaluation of a commercial bovine viral diarrhea virus vaccine in nonpregnant female alpacas (Vicugna pacos).

Bovine viral diarrhea virus (BVDV) is an emerging pathogen in alpacas and many questions still persist regarding disease mechanisms and control strategies. The purpose of this study was to evaluate a commercial BVDV vaccine for safety and efficacy in alpacas. Five nonpregnant alpacas were vaccinated with a modified-live BVDV vaccine and challenged 25 days post-immunization by nasal and ocular inoculation with a BVDV Type 1b strain isolated from a confirmed BVDV persistently infected alpaca. Two nonpregnant alpacas served as non-vaccinated controls and were similarly challenged. Results indicated that BVDV virus could not be detected from the vaccinated alpacas but was detected in the unvaccinated alpacas. Results suggest that administration of modified-live BVDV vaccine protected the alpacas in this study from experimental challenge and no adverse effects from the vaccine were observed.

Development of a bovine ileal cannulation model to study the immune response and mechanisms of pathogenesis of paratuberculosis.

An ileal cannulation model was developed in conjunction with a flow cytometric assay to gain a better understanding of the mechanisms of immunopathogenesis of Johne's disease caused by Mycobacterium avium subsp. paratuberculosis. Initial studies with calves showed that M. avium subsp. paratuberculosis DNA is detectable by PCR in ileal biopsies during the first months following experimental infection. Inflammatory lesions were not detected on endoscopic evaluation up to 8 months postexperimental infection. M. avium subsp. paratuberculosis DNA was detected in multiple tissues at necropsy 8 months postinfection. Examination of the activation status of epithelial lymphocytes from the jejunum and ileum from infected and control animals at necropsy revealed that none of the major subsets of lymphocytes (NK, CD2(+), and CD2(-) gammadelta T lymphocytes, or CD4 and CD8 alphabeta T lymphocytes) expressed activation molecules CD25, CD26, CD71, ACT1, or ACT16. Subsets of CD4 and CD8 T lymphocytes from control and infected animals expressed CD26. The majority of CD4 and CD8 T lymphocytes expressed CD45R0, the memory T-lymphocyte marker. An immune response to M. avium subsp. paratuberculosis was detected by 3 months postinfection, dominated by a strong proliferative response of CD4 memory T lymphocytes. The findings indicate an immune response develops following initial exposure to M. avium subsp. paratuberculosis that controls but does not eliminate the pathogen. This persistence of M. avium subsp. paratuberculosis possibly leads to erosion and dysregulation of protective immunity at later time points postinfection. Continuous access to the ileum offers an opportunity to elucidate the cellular and molecular events leading to immune dysregulation and development of chronic inflammatory ileitis.

Disseminated Bovine viral diarrhea virus in a persistently infected alpaca (Vicugna pacos) cria.

Bovine viral diarrhea virus (BVDV) is an emerging infectious pathogen of concern to the alpaca industry. A 4-month-old, intact, male alpaca cria was diagnosed as persistently infected with BVDV on the basis of repeated positive antemortem polymerase chain reaction (PCR) and virus isolation (VI) assays and negative serologic titers to BVDV. Immunohistochemistry, real-time reverse transcription PCR, and VI performed on tissues collected at necropsy demonstrated disseminated BVDV-1b infection. Virus was detected in multiple tissues, including parotid salivary gland, testes, prostate, kidneys, skin, and gastrointestinal tract. Demonstration of BVDV in previously unreported tissues suggests additional potential routes of BVDV transmission in alpacas.

Physiologic mastectomy via flank laparotomy.

Physiologic mastectomy can be used as a salvage procedure in cases of chronic suppurative mastitis, gangrenous mastitis, or chronic, severe mastitis associated with organisms liberating endotoxin or exotoxin. The surgical technique involves ligation of the major arterial blood supply (external pudendal artery) to the corresponding half of the mammary gland, which results in decreased systemic absorption of toxins and gland atrophy. The technique is performed with the cow standing, and it is relatively atraumatic. This procedure is a simple, yet effective alternative to radical mastectomy for unresponsive mastitis cases in genetically or otherwise valuable cattle.

A fungal granuloma of the frontal sinus in a llama.

A 12-year-old, castrated male llama (Lama glama) presented with a 12-cm diameter cranial mass. Computed tomography and postmortem examination revealed that the mass invaded the calvarium and compressed the rostral part of the brain. Light microscopic examination confirmed a fungal granuloma.

Use of flow cytometry to identify monoclonal antibodies that recognize conserved epitopes on orthologous leukocyte differentiation antigens in goats, llamas, and rabbits.

Flow cytometry was used to screen a panel of 320 mAbs, submitted to the Animal Homologues Section of the HLDA8, for mAbs that recognize epitopes conserved on orthologous leukocyte differentiation antigens (LDA) in goats, lamas, and rabbits. Nineteen mAbs specific for CD11a (1), CD14 (3), CD18 (1), CD21 (1), CD29 (2), CD44 (2), CD47 (3), CD49d (1), CD172a (1), CD45RB (1), CD61 (1), RACT48A, and GBSP71A reacted with goat LDA. Twenty three mAbs specific for CD7 (1), CD9 (2), CD11a (1), CD14 (3), CD18 (4), CD29 (1), CD32 (1), CD44 (1), CD47 (4), CD49d (2), CD50 (1), CD80 (1), CD172a (1), and GBSP71A reacted with llama LDA. Eighteen mAbs specific for CD9 (2), CD11a (1), CD14 (2), CD18 (4), CD21 (1), CD44 (2), CD45RB (1), CD49d (1), CD209 (1), RACT48A, and GBSP71A reacted with rabbit LDA. The specificities of two cross reactive mAbs that recognize different conserved epitopes on all leukocytes in two species (RACT48A) and all three species (GBSP71A) have not been determined. The patterns of reactivity of most of the mAbs were consistent with patterns of reactivity noted on human leukocytes. The specificity of some cross reactive mAbs generated in non-human species were validated on human leukocytes. Further studies are needed to verify that CD7, CD32, CD45RB, CD50, and CD209 recognize orthologous molecules in the indicated species.

Comparison of disease severity scoring systems for dairy cattle with acute coliform mastitis.

OBJECTIVE: To compare use of 4 disease severity scoring systems to predict bacteremia (yes vs no) and outcome (survived vs died or culled) in dairy cows with acute coliform mastitis (ACM). DESIGN: Retrospective cohort study. ANIMALS: 99 dairy cows with ACM. PROCEDURES: Cows were classified as having mild, moderate, or severe disease with a scoring system based on systemic disease signs alone (systemic severity score [SSS] system), a system based on local disease signs alone (local severity score [LSS] system), and 2 previously described systems based on a combination of local and systemic signs (local-systemic score 1 [LS1] and local-systemic score 2 [LS2] systems). Test performance was calculated to determine whether a severe disease classification could be used to predict bacteremia or outcome. RESULTS: 21%, 53%, 63%, and 38% of cows were classified as having severe disease with the SSS, LSS, LS1, and LS2 systems, respectively. For both bacteremia and outcome, sensitivity was highest for the LS1 system, but specificity and accuracy were highest for the SSS system. Examination of a scatterplot of true-positive rate versus false-positive rate for each of the scoring systems indicated that the SSS and LS2 systems were similar in their ability to correctly identify cows with bacteremia or an adverse outcome. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the SSS scoring system was better for identifying cows with bacteremia or an adverse outcome than was the LSS system and that the LS1 and LS2 systems were intermediate in their discriminatory abilities.

Use of rMPB70 protein and ESAT-6 peptide as antigens for comparison of the enzyme-linked immunosorbent, immunochromatographic, and latex bead agglutination assays for serodiagnosis of bovine tuberculosis.

Current assays used to detect Mycobacterium bovis infection lack accuracy, especially for recently infected animals, or are impractical for rapid field diagnostic applications. To overcome these limitations with serological assays, a synthetic peptide derived from early secretory antigenic target 6 (ESAT6-p) and a recombinant major secreted immunogenic protein (rMPB70) of M. bovis were used in an enzyme-linked immunosorbent assay (EIA), an immunochromatographic assay (ICGA), and a latex bead agglutination assay (LBAA). Sera from noninfected, M. bovis-infected, or M. avium subsp. paratuberculosis-infected (by natural and experimental routes) animals were evaluated. Receiver operating characteristic analysis comparing optical density values from the EIA with results of bacterial culture or skin test, the reference test, established suitable cutoff values for assessing sensitivity and specificity. The EIA and LBAA, respectively, had sensitivities of 98.6 and 94.8%, specificities of 98.5 and 92.6%, and kappa values of 0.97 and 0.88 with ESAT6-p. The EIA, ICGA, and LBAA, respectively, had sensitivities of 96.8, 83.0, and 86.7%, specificities of 90.1, 99.4, and 97.8%, and kappa values of 0.87, 0.85, and 0.83 with rMPB70. Examination of serial samples of sera collected from experimentally M. bovis-infected cattle and deer revealed that ESAT6-p-specific responses developed early after infection whereas responses to rMPB70 developed later in the course of disease. The advantage of the LBAA and ICGA as initial tests for multiple species is a rapid reaction obtained in 2 to 3 h by LBAA or 20 min by ICGA without species-specific secondary antibodies under field conditions, thus allowing immediate segregation of suspect animals for further testing before culling.