Bovine botulism is a fatal disease that is caused by botulinum neurotoxins (BoNTs) produced by Clostridium botulinum serotypes C and D and that causes great economic losses, with nearly 100% ...lethality during outbreaks. It has also been considered a potential source of human food-borne illness in many countries. Vaccination has been reported to be the most effective way to control bovine botulism. However, the commercially available toxoid-based vaccines are difficult and hazardous to produce. Neutralizing antibodies targeted against the C-terminal fragment of the BoNT heavy chain (HC) are known to confer efficient protection against lethal doses of BoNTs. In this study, a novel recombinant chimera, consisting of Escherichia coli heat-labile enterotoxin B subunit (LTB), a strong adjuvant of the humoral immune response, fused to the HC of BoNT serotypes C and D, was produced in E. coli. Mice vaccinated with the chimera containing LTB and an equivalent molar ratio of the chimera without LTB plus aluminum hydroxide (Al(OH)3) developed 2 IU/mL of antitoxins for both serotypes. Guinea pigs immunized with the recombinant chimera with LTB plus Al(OH)3 developed a protective immune response against both BoNT/C (5 IU/mL) and BoNT/D (10 IU/mL), as determined by a mouse neutralization bioassay with pooled sera. The results achieved with guinea pig sera fulfilled the requirements of commercial vaccines for prevention of botulism, as determined by the Brazilian Ministry of Agriculture, Livestock and Food, Supply. The presence of LTB was essential for the development of a strong humoral immune response, as it acted in synergism with Al(OH)3. Thus, the vaccine described in this study is a strong candidate for the control of botulism in cattle.
The heat-labile enterotoxin of Escherichia coli (LT) consists of an enzymatically active A subunit (LTA) and a pentameric B subunit (LTB). LT has been extensively studied as a potent modulator of ...immune responses but wild-type LT is toxic and therefore unsuitable for clinical use. Approaches pursued to avoid the toxicity associated with the use of the native toxin while retaining its adjuvant properties have included isolation of subunit B (LTB) and construction of non-toxic LT AB complex mutants, such as LTK63 mutant. Here we review the immunomodulatory characteristics of LTB and LTK63 and their potential as mucosal and parenteral vaccine adjuvants.
Brazilian ethanol production generates approximately 351 billion liters yearly of vinasse that is mostly used for fertigation, but with significant environmental impact. This research aims at ...comparing the growth of
Chlorella vulgaris
on three different media preparation using raw vinasse, diluted vinasse, and vinasse treated by anaerobic digestion; all under four distinct treatments or supplementations (antibiotic, nitrogen and phosphorous). The results show that
C vulgaris
grows poorly on raw vinasse alone or after supplementation. However, 11–13 mg L
−1
day
−1
of algal oil can be produced with diluted or anaerobically treated vinasse. The first would require antibiotic, nitrogen and phosphorous supplementation and the later only antibiotic supplementation. Whilst using diluted vinasse may reduce work input, it requires significant amounts of clean water. The use of anaerobically treated vinasse requires some additional work and investment, but it can generate 122 mL CH
4
g
−1
COD
vinasse
(10–26.4 m
3
of biogas from 1m
3
of vinasse). Thus, the association of methane and algal oil production from vinasse is an effective approach for removing inorganic nutrients and potentially toxic organics from this effluent before discharge. The quality of the algal oil produced is affected by culturing conditions and anaerobically treated vinasse produced the best results in this area according to the estimated parameters of the biodiesel.
Immunomagnetic separation (IMS) and immunoassays are widely used for pathogen detection. However, novel technology platforms with highly selective antibodies are essential to improve detection ...sensitivity, specificity and performance. In this study, monoclonal antibodies (MAbs) against Internalin A (InlA) and p30 were generated and used on paramagnetic beads of varying diameters for concentration, as well as on fiber-optic sensor for detection.
Anti-InlA MAb-2D12 (IgG2a subclass) was specific for Listeria monocytogenes and L. ivanovii, and p30-specific MAb-3F8 (IgM) was specific for the genus Listeria. At all bacterial concentrations (10³-10⁸ CFU/mL) tested in the IMS assay; the 1-μm diameter MyOne beads had significantly higher capture efficiency (P < 0.05) than the 2.8-μm diameter M-280 beads with both antibodies. The highest capture efficiency for MyOne-2D12 (49.2% for 10⁵ CFU/mL) was significantly higher (P < 0.05) than that of MyOne-3F8 (16.6 %) and Dynabeads anti-Listeria antibody (9 %). Furthermore, capture efficiency for MyOne-2D12 was highly specific for L. monocytogenes and L. ivanovii. Subsequently, we captured L. monocytogenes by MyOne-2D12 and MyOne-3F8 from hotdogs inoculated with mono- or co-cultures of L. monocytogenes and L. innocua (10-40 CFU/g), enriched for 18 h and detected by fiber-optic sensor and confirmed by plating, light-scattering, and qPCR assays. The detection limit for L. monocytogenes and L. ivanovii by the fiber-optic immunosensor was 3 × 10² CFU/mL using MAb-2D12 as capture and reporter antibody. Selective media plating, light-scattering, and qPCR assays confirmed the IMS and fiber-optic results.
IMS coupled with a fiber-optic sensor using anti-InlA MAb is highly specific for L. monocytogenes and L. ivanovii and enabled detection of these pathogens at low levels from buffer or food.
Bovine herpesviruses 1 and 5 (BoHV-1 and BoHV-5) are economically important pathogens, associated with a variety of clinical syndromes, including respiratory and genital disease, reproductive failure ...and meningoencephalitis. The standard serological assay to diagnose BoHV-1 and BoHV-5 infections is the virus neutralization test (VNT), a time consuming procedure that requires manipulation of infectious virus. In the present study a highly sensitive and specific single dilution indirect ELISA was developed using recombinant glycoprotein D from BoHV-5 as antigen (rgD5ELISA). Bovine serum samples (n = 450) were screened by VNT against BoHV-5a and by rgD5ELISA. Compared with the VNT, the rgD5ELISA demonstrated accuracy of 99.8%, with 100% sensitivity, 96.7% specificity and coefficient of agreement between the tests of 0.954. The rgD5ELISA described here shows excellent agreement with the VNT and is shown to be a simple, convenient, specific and highly sensitive virus-free assay for detection of serum antibodies to BoHV-5.
Display omitted
The objective of this observational study was to evaluate the efficacy of a neck-mounted automated activity monitor (AAM) at detecting early postpartum resumed ovarian cyclicity. A ...total of 192 lactating cows (primiparous = 73 and multiparous = 119) were enrolled in this study. Cows were continuously monitored by a neck-mounted AAM early postpartum (7 to 30 d in milk; DIM). Calving was classified as assisted (forced extraction of a calf) or unassisted (normal calving). Retained fetal membrane, metritis, hyperketonemia, clinical mastitis, and milk production were recorded. Cows were classified as healthy (i.e., no disease events) or sick (i.e., any disease event). Estrus events were alerted by the AAM using a proprietary algorithm set by the AAM company. Blood samples, from the coccygeal vein, were collected at 15, 18, 21, 24, 28, and 30 DIM for progesterone (P4) analysis. Resumption of cyclicity was considered when P4 concentration was ≥1 ng/mL on any collection day. Cows were considered anovular when P4 concentration was <1 ng/mL on all collection days. Cows were classified as true positive: P4 ≥ 1 ng/mL and at least one estrus alert; false positive: P4 < 1 ng/mL and at least one estrus alert; true negative: P4 < 1 ng/mL and no estrus alerts; and false negative: P4 ≥ 1 ng/mL and no estrus alerts. Statistical analyses were performed by frequency distribution and mixed effects logistic regression procedures on SAS (SAS Institute Inc.). The specificity, sensitivity, accuracy, and positive predictive value of the sensor to detect cows that had resumed cyclicity were 84.0%, 34.1%, 52.1%, and 79.2%, respectively. Out of the 192 cows, 35.9% (69/192) were anovulatory and 37.5% (72/192) had no estrus events between 7 to 30 DIM. Healthy cows were more likely to resume cyclicity in early lactation compared with cows that were sick (78.3 ± 1.9 vs. 32.8 ± 3.1%, respectively) independent of parity. In conclusion, the sensor had a high specificity for detecting anovular cows, but it had lower sensitivity, and thus was not effective at detecting cyclic cows, perhaps due to silent ovulation early postpartum.
PDF
