Dietary fiber source and direct-fed microbial supplementation effects on lactation performance and feeding behavior of high-producing dairy cows.

The objective of this study was to evaluate the effects of dietary fiber source and direct-fed microbial supplementation on lactation performance and feeding behavior of high-producing dairy cows. Sixty-four multiparous Holstein cows (3.5 ± 1.6 lactations; 76 ± 22 DIM and 735 ± 67 kg of BW at covariate period initiation) and 32 gate feeders were enrolled in a study with a completely randomized design and a 2 × 2 factorial arrangement. Cows and gate feeders were randomly assigned to treatments (16 cows and 8 gate feeds per treatment). Cows were allowed 1 week to acclimate to feeding gates followed by a 2-week covariate period. During the acclimation and covariate periods, all cows were fed the same diet to meet or exceed the nutrient requirements. Following the covariate period, cows were enrolled in a 8-week treatment period during which cows were randomly assigned to 1 of 4 treatments consisting of forage inclusion in the diet, either 45.8% (LF) or 56.7% (HF) of DM, and the supplementation of 75 mL/hd/d of a direct-fed microbial (DFM) containing herbal extracts (mallow, mint, and sage), L. plantarum, L. buchneri, Saccharomyces cerevisiae, and sugar cane molasses (Valibiom Mix, Valibiotics, Traiskirchen, Austria) or without supplementation (CON). The average covariate period value of each variable was used as a covariate. Three-way interactions were observed for DMI and feed efficiency. Dry matter intake was 2 to 3 kg greater for LF-DFM than HF-CON and HF-DFM during wk 2, 3, 5, and 8 of the treatment period. Milk production was 2.1 kg/d greater for LF than HF diets. Both milk fat and MUN concentrations were greater for HF than LF diets. Conversely, milk protein concentration was lower for HF than LF diets. The respiration rate measured in the morning was lower with DFM supplementation than CON. Rectal temperature measured in the morning and averaged for the day were greater for LF than HF diets. Under the conditions of the present study, feeding high-forage diets may be an alternative for producers to reduce feeding costs depending on the price of purchased feeds. However, non-forage fiber sources (i.e., soy hulls) must be considered when producers are challenged by either forage shortages or forage with a lower nutritive value. Additionally, DFM supplementation reduced respiration rate in the morning and affected meal behavior of lactating cows.

Effects of a genetically modified corn hybrid with α-amylase and storage length on fermentation profile and starch disappearance of whole-plant corn silage and earlage.

Two experiments were conducted to evaluate the effects of a genetically modified corn hybrid with α-amylase expressed in the kernel (AMY) on fermentation profile, aerobic stability, nutrient composition, and starch disappearance of whole-plant corn silage (WPCS) and earlage. Both hybrids, AMY and an isogenic corn hybrid (ISO), were grown in 10 replicated plots (5 for WPCS and 5 for earlage). Samples of each plot were collected at harvest, homogenized, and divided into 5 subsamples which were randomly assigned to 5 storage lengths (0, 30, 60, 90, and 120 d). Both datasets (WPCS and earlage), were analyzed separately as a completely randomized block design in a factorial arrangement of treatments, with a model including the fixed effects of hybrid, storage length, and their interaction, and the random effect of block. Minor differences on fermentation profile were observed between AMY and ISO for WPCS and earlage. An interaction between hybrid and storage length was observed for DM losses in WPCS, where losses were similar at 30, 60 and 90 d, but lower for AMY compared with ISO at 120 d. No effect of hybrid was observed on yeast and mold counts for WPCS or earlage. The aerobic stability of WPCS was greater for AMY than ISO. For earlage, AMY had greater DM losses and aerobic stability than ISO. An interaction between hybrid and storage length was observed for ammonia-N in both WPCS and earlage, where ammonia-N was similar at 0 d but greater for AMY than ISO throughout later storage lengths. A similar interaction was observed for water-soluble carbohydrates (WSC) concentrations in WPCS, where ISO had greater WSC than AMY at 0 d but was similar throughout later storage lengths. However, AMY earlage had a greater WSC concentration throughout storage length, but a lesser magnitude after ensiling. Starch concentration was greater for AMY than ISO in WPCS and earlage. Greater starch disappearances at 0 h and 6 h were observed for ISO in WPCS and earlage. Minor effects on fermentation profile, microbial counts, aerobic stability and nutrient composition suggests that AMY can be ensiled for prolonged periods with no concerns for undesirable fermentation or nutrient losses. However, in situ starch disappearance was lower for AMY compared with ISO.

Effects of season, variety type, and trait on dry matter yield, nutrient composition, and predicted intake and milk yield of whole-plant sorghum forage.

Sorghum forage is an important alternative to high-quality forage in regions where climatic and soil conditions are less desirable for corn production for silage and producing comparable nutritive value is challenging. The objective of this experiment was to assess the effects of season (spring vs. summer), sorghum variety type (forage sorghum vs. sorghum-sudangrass), and trait [brown midrib (BMR) vs. non-BMR] on dry matter (DM) yield, nutrient composition, and predicted intake and milk yield of whole-plant sorghum forage grown in Florida from 2008 to 2019. Whole-plant sorghum forage was harvested at a targeted 32% of DM, and each year, spring (April) and summer (July) trials were established. A total of 300 forage sorghum and 137 sorghum-sudangrass hybrids were tested for a total of 437 hybrids, of which 199 hybrids contained the BMR trait and 238 were non-BMR. An interaction between season and sorghum variety type was observed for DM yield. Dry matter yield was greater for the spring season than the summer season, with sorghum-sudangrass outperforming forage sorghum only during the spring season. In addition, BMR hybrids had a lower DM yield than non-BMR hybrids, regardless of season and variety type. An interaction between season and trait was observed for predicted neutral detergent fiber digestibility after 30 h of incubation in rumen fluid (NDFD30h). Predicted NDFD30h was greater for BMR sorghum in comparison to non-BMR sorghum, but BMR sorghum had slightly greater predicted NDFD30h when grown in the spring than summer, whereas no seasonal differences were found for predicted NDFD30h across non-BMR sorghum. An interaction between season, variety type, and trait was observed for predicted dry matter intake at 45 (DMI45), 55 (DMI55), and 65 (DMI65) kg of milk/d. Predicted DMI45 and DMI55 were greater for spring BMR forage sorghum than for spring and summer non-BMR sorghum-sudangrass and were greater for spring BMR forage sorghum than for summer BMR sorghum-sudangrass. Predicted DMI65 was greater for BMR forage sorghum in comparison to all non-BMR hybrids in the spring. Additionally, spring BMR forage sorghum was greater than summer sorghum-sudangrass regardless of trait. An interaction between season and sorghum variety type was observed for milk yield per megagram of forage. Milk yield per megagram of forage was greatest for spring forage sorghum. Sorghum variety type and trait selection are crucial to minimize differences in forage nutritive value of sorghum forage between seasons and improve the performance of high-producing dairy cows.

Effects of chemical additive and packing density on the fermentation profile and nutrient composition of ensiled cocktail forage mix.

Recently, the use of cocktail forage mixes in dairy cattle rations has become more common because the mixtures are low-cost, fit well in rotation after a cereal grain forage, and can have similar yield and energy value compared with alfalfa silage. This experiment evaluated the effects of a chemical additive and packing density on the fermentation profile and nutrient composition of cocktail mix silage. The cocktail forage mix (brown-midrib sorghum-sudangrass, Italian ryegrass, red clover, berseem clover, and hairy vetch) was harvested, ensiled in laboratory silos (3.79-L plastic buckets), and allowed to ferment for 30 d. The experiment consisted of 6 treatments, 2 chemical additives [CON (30 mL of distilled water) or ADD (sodium sulfite, sodium metabisulfite, and fungal amylase)], and 3 packing densities [D100, D75, and D50 (100%, 75%, or 50% of the maximum material in laboratory silos, respectively)], for a total of 24 silos (4 replications per treatment combination). No interactions of additive by density were detected for any parameters evaluated. The addition of the chemical additive influenced fermentation profile, with reduced concentrations of total acids, lactic acid, acetic acid, and ethanol in ADD-treated silages. Moreover, D50 reduced concentrations of total acids, lactic acid, and acetic acid compared with D100, but had greater pH and yeast and mold counts. Minimal changes in nutrient composition were detected regardless of treatment. Overall, this study corroborates the importance of a well-packed silage during the ensiling process. Poorly packed cocktail mix silages may be more prone to spoilage based on yeast and mold counts.

Repeated dose (14 days) rat intramuscular toxicology study of Her1 vaccine.

Our goal was to assess the toxicity of two strengths (200 and 400 µg) of HER1 cancer vaccine (Center of Molecular Immunology, Cuba), presented in two different formulations, in Sprague Dawley rats after repeated intramuscular administration (14 days). Four groups (5 animals/sex) were established: Control, Placebo (adjuvant), and two Treated groups receiving a dose representing ten times of human total dose (10×), 28.6 and 57.1 µg/kg. Clinical observations, body weight and rectal temperature were measured during the study. Clinical pathology analysis was performed, besides gross necropsy and histological examination of tissues on animals at the end of the assay. The assay ended with a 100% survival. Injection site damage, with the presence of cysts and granulomas, was observed in adjuvant and vaccine treated groups, with most severe cases predominating at higher strength. Administration of Placebo and Her1 vaccine induced increase in polymorphonuclear cells, with relative lymphopenia conditioned by primary neutrophilia. In summary, results suggest that Her1 immunization was capable of inducing an inflammatory effect at the injection site, leading to systemic alterations, more significant at higher strength (400 µg, 57.1 µg/kg), probably affected by the immunizations' schedule used. The vaccine was shown to be well tolerated without any obvious signs of systemic toxicity, with findings largely attributable to the adjuvant used.

Acute oral, pulmonary and intravenous toxicity/pathogenicity testing of a new formulation of Bacillus thuringiensis var israelensis SH-14 in rats.

During the last decades, efforts are being made to develop microbial insecticides as biological control agents. Bacillus thuringiensis has been one of the most consistent and significant biopesticides for using on crops as an insecticidal spray. The aim of this study was to assess and to compare the pathogenicity of a new formulation of B.thuringiensis var israelensis SH-14 in rats through oral, intranasal and intravenous single dosing. Through 21 days after administration, clinical examinations were performed daily, and body weight gain was evaluated. Clearance was estimated by means of collection of feces or examination of lungs and blood, and infectivity was evaluated enumerating microorganisms from organs of Bti SH-14 treated animals sacrificed at intervals. Gross necropsy of animals was performed at interim or final sacrifice. There were no treatment-related mortalities, and no evidence of pathogenicity or treatment related toxicity, although in the intravenous study, the microorganism was capable of achieving persistence in organs after administration, and the Bti SH-14 treated animals developed skin ulcerations and hemorrhages at the injection site. It could be concluded that the tested microorganism was not toxic or pathogenic to rats via oral or intranasal route, although it was capable of achieving persistence in organs after intravenous administration, eliciting local effects at the injection site.

Kinetics of antibodies in sera, saliva, and urine samples from adult patients with primary or secondary dengue 3 virus infections.

OBJECTIVES: The kinetics of three serological markers (IgM, IgA, and IgG) in serum, saliva, and urine samples from adult patients with primary or secondary dengue infection were studied. DESIGN: Serum, saliva, and urine samples were collected from 22 patients with clinical and confirmed dengue 3 virus infection during the outbreak in Havana City in 2001. They were tested by capture IgM (MAC-ELISA), IgA (AAC-ELISA), and IgE (EAC-ELISA) and IgG ELISA inhibition method (EIM) to detect specific dengue antibodies. RESULTS: Similar kinetics were observed in IgM, IgA, and IgG antibodies in saliva and IgA and IgG in urine samples from secondary cases compared with kinetics in serum samples, although the values were lower. No IgG antibody was detected in saliva and urine samples in primary cases and IgM antibody was not detected in urine samples from either primary or secondary infection. All secondary cases were positive for IgG in saliva and urine samples at day 7. The kinetics of specific IgE antibodies in primary and secondary cases were different. CONCLUSIONS: The kinetics of three serological markers (IgM, IgA, and IgG) in serum, saliva, and urine samples from adult patients with primary or secondary dengue 3 virus infection were studied for the first time, showing its behavior and usefulness in dengue virus diagnosis. The specific IgE could play a role as a serological marker in secondary infections.

Gas-phase fragmentation of protonated piplartine and its fungal metabolites using tandem mass spectrometry and computational chemistry.

Piplartine, an alkaloid produced by plants in the genus Piper, displays promising anticancer activity. Understanding the gas-phase fragmentation of piplartine by electrospray ionization tandem mass spectrometry can be a useful tool to characterize biotransformed compounds produced by in vitro and in vivo metabolism studies. As part of our efforts to understand natural product fragmentation in electrospray ionization tandem mass spectrometry, the gas-phase fragmentation of piplartine and its two metabolites 3,4-dihydropiplartine and 8,9-dihydropiplartine, produced by the endophytic fungus Penicillium crustosum VR4 biotransformation, were systematically investigated. Proposed fragmentation reactions were supported by ESI-MS/MS data and computational thermochemistry. Cleavage of the C-7 and N-amide bond, followed by the formation of an acylium ion, were characteristic fragmentation reactions of piplartine and its analogs. The production of the acylium ion was followed by three consecutive and competitive reactions that involved methyl and methoxyl radical eliminations and neutral CO elimination, followed by the formation of a four-member ring with a stabilized tertiary carbocation. The absence of a double bond between carbons C-8 and C-9 in 8,9-dihydropiplartine destabilized the acylium ion and resulted in a fragmentation pathway not observed for piplartine and 3,4-dihydropiplartine. These results contribute to the further understanding of alkaloid gas-phase fragmentation and the future identification of piplartine metabolites and analogs using tandem mass spectrometry techniques. Copyright © 2017 John Wiley & Sons, Ltd.

Serological markers during dengue 3 primary and secondary infections.

BACKGROUND: The detection of the IgM antibody for the dengue virus in serum by ELISA has become one of the most important and useful methods for diagnosis of dengue using a single acute-phase serum sample. Currently, this system is an invaluable tool for the surveillance of dengue fever (DF) and dengue hemorrhagic fever (DHF). The usefulness of other serological markers such as IgA and IgE have been less studied. OBJECTIVE: To study the IgM, IgA and IgE specific antibody response in dengue 3 infected patients with different clinical picture and type of infection. STUDY DESIGN: One hundred and twenty-seven serum samples collected on days 5-7 at the onset of fever from clinically and serologically confirmed dengue cases were studied. Forty-two were classified as primary dengue fever cases, 48 as secondary dengue fever cases and 37 as secondary dengue hemorrhagic fever cases. All samples were tested by capture ELISA in order to detect dengue IgM, IgA and IgE antibodies. RESULTS AND CONCLUSIONS: In this study, significant differences were observed in the IgM, IgA and IgE response between the study groups. High IgA and IgE OD ratios in secondary dengue cases were found. The usefulness of serotype specific IgM antibody detection is also analyzed and discussed. A priority for future dengue research in terms of protection, recovery of infection and immunopathogenesis is to elucidate the role of these immunoglobulins. The cross reactivity response to IgM between dengue virus serotypes in primary and secondary cases should also be more studied.

[Spectral analysis of the electroencephalogram in children with systemic lupus erythematosus]. / Análisis espectral del electroencefalograma en niños con lupus eritematoso sistémico.

INTRODUCTION: Systemic lupus erythematosus (SLE) is an autoimmune disease that affects different systems and organs, including the central nervous system (CNS). It has been suggested that about 40% of the cases of neuropsychiatric lupus (NPSLE) develop before SLE is diagnosed or at the same time it is being carried out, and 63% appear during the first year following diagnosis. AIMS: The aim of this study was to check the hypothesis that the electroencephalogram (EEG) may be sensitive to the damage to the CNS in children with SLE in whom there is still no clinical evidence of NPSLE. PATIENTS AND METHODS: EEG recordings were performed in 30 children with a diagnosis of SLE with or without signs of a neuropsychiatric syndrome. The results of the EEG were evaluated visually and analysed quantitatively. RESULTS: The visual inspection of the EEG showed the presence of alterations in 44.5% of the children with SLE and in 76.9% of those with NPSLE. There were significant differences in Student's t test (p = 0.0055) between the two groups for the analysis of the broadband spectral measurements. The narrow band analysis revealed a significant increase in the theta and delta frequencies in children with SLE as compared to standard values, whereas in children with NPSLE significant differences were found in the fast bands in frontal regions. CONCLUSIONS: Spectral analysis of the narrow band could help to confirm diagnoses of NPSLE, while anomalies in the slow bands could be an early marker of damage to the CNS although there are still no symptoms of the disease.

Structure of Trypanosoma cruzi glycosomal glyceraldehyde-3-phosphate dehydrogenase complexed with chalepin, a natural product inhibitor, at 1.95 A resolution.

The structure of the glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) from Trypanosoma cruzi complexed with chalepin, a natural product from Pilocarpus spicatus, has been determined by X-ray crystallography to 1.95 A resolution. The structure is in the apo form without cofactors in the subunits of the tetrameric gGAPDH in the asymmetric unit. Unequivocal density corresponding to the inhibitor was clearly identified in one monomer. The final refined model of the complex shows extensive conformational changes when compared with the native structure. The mode of binding of chalepin to gGAPDH and its implications for inhibitor design are discussed.

Pyrano chalcones and a flavone from Neoraputia magnifica and their Trypanosoma cruzi glycosomal glyceraldehyde-3-phosphate dehydrogenase-inhibitory activities.

The fruits of Neoraptua magnifica var. magnifica afforded three new flavonoids: 2'-hydroxy-4,4',-dimethoxy-5',6'-(2'',2''-dimethylpyrano)chalcone, 2'-hydroxy-3,4,4'-trimethoxy-5',6'-(2'',2''-dimethylpyrano)chalcone, and 3',4'-methylenedioxy-5,7-dimethoxyflavone which were identified on the basis of spectroscopic methods. The known flavonoids 2'-hydroxy-3,4,4',5-tetramethoxy-5',6'-(2'',2''-dimethylpyrano)chalcone, 2'-hydroxy-3,4,4',5,6'-pentamethoxychalcone, 3',4'-methylenedioxy-5,6,7-trimethoxyflavone, 3',4'-methylenedioxy-5',5,6,7-tetramethoxyflavone, 3',4',5',5,7-pentamethoxyflavanone and 3',4',5'5,7-pentamethoxyflavone were also identified. The latter flavone was the most active as glyceraldehyde-3-phosphate dehydrogenase-inhibitor.

Study of biologic attributes of Cuban dengue 2 virus after serial passage in primary dog kidney cells.

OBJECTIVE: The serial passage of dengue viruses in primary dog kidney (PDK) cells has resulted in selection of attenuated viruses. However, the molecular changes responsible for loss of virulence are not well characterized. This article describes the isolation and biologic attributes of one dengue 2 virulent strain as a first step to allow the study of determinants of virulence at the molecular level. METHODS: A15 dengue 2 Cuban strain was isolated from the viremic plasma of a patient with uncomplicated dengue fever during the 1981 epidemic. This was then subjected to serial passage in PDK cells. Viruses resulting from several PDK passages were compared to the parent strain for plaque size and temperature sensitivity, neurovirulence in newborn mice, and cytopathogenic effects on LLC-MK(2) and C6/36-HT cell lines. RESULTS: A15 dengue 2 Cuban strain was successfully propagated in PDK cells. Primary dog kidney 52 to 53 viruses exhibited several biologic attributes, such as small plaques, temperature sensitivity, reduced mouse neurovirulence, and cytopathic effect in permissive cell lines. CONCLUSIONS: These results represent the first step to allow attenuation of this strain of dengue 2 virus.

Quantitative ELISA for mouse monoclonal antibody determination in culture supernatants and in human serum.

An ELISA has been set up for quantifying mouse monoclonal antibodies in culture supernatant. The assay includes rabbit anti-mouse IgG antibodies chromatographycally purified. This preparation was used as coating and as conjugated antibodies in the ELISA. The assay can detect IgG1 with sensitivity of 0.2 ng/mL, IgG2a (0.85 ng/mL), IgG2b (0.13 ng/mL), and IgG3 (3.19 ng/mL) in culture supernatants. The effective working range was from subnanogram per mL quantities to 30 ng/mL by using a computer statistical program. Variation coefficient of ELISA was below 7%. Correlation estimates with a similar ELISA using commercial reagents were performed for each mouse antibody subclass. The assay was able to detect the four mouse monoclonal antibody subclasses in pure human serum as compared with the same ELISA using commercial antibodies. A 24-h pharmacokinetic profile of 1 patient treated with an IgG2a monoclonal antibody is presented.

Two novel GPER agonists induce gene expression changes and growth effects in cancer cells.

Although the action of estrogens has been traditionally explained by the binding to and transactivation of the nuclear estrogen receptor (ER)α and ERß, recently the G protein-coupled receptor GPR30/GPER has been involved in the rapid estrogen signaling. We investigated the ability of two original molecules, which were named GPER-L1 and GPERL2, to bind to and activate the GPER transduction pathway in cancer cells. Competition assays, docking simulations, transfection experiments, real-time PCR, immunoblotting, gene silencing technology and growth assays were performed to ascertain the selective action of GPER-L1 and GPER-L2 in activating the GPER-mediated signaling. Both compounds, which did not show any ability to bind to and activate the classical ERs, were able to bind to GPER and to trigger the rapid activation of the GPER/EGFR/ERK transduction pathway which led to the up-regulation of GPER-target genes. Notably, GPER-L1 and GPER-L2 induced the proliferation of SkBr3 breast and Ishikawa endometrial cancer cells at nM concentrations through GPER, hence providing further evidence on their capability to elicit relevant biological responses mediated by GPER. The identification and characterization of these novel compounds as selective GPER agonists represent a valuable tool to further dissect the pharmacology of this novel estrogen receptor and to better differentiate the specific functions elicited by each estrogen receptor subtype in cancer cells.

Δ-lactam derivative from thermophilic soil fungus exhibits in vitro anti-allergic activity.

From cultures of thermophilic soil fungus Humicola grisea var thermoidea, a δ-lactam derivative (3-(2-(4-hydroxyphenyl)-2-oxoethyl)-5,6-dihydropyridin-2(1H)-one) that displayed anti-allergic activity was isolated, which was predicted by in silico computational chemistry approaches. The in vitro anti-allergic activity was investigated by ß-hexosaminidase release assay in rat basophilic leukaemia RBL-2H3 cells. The δ-lactam derivative exhibited similar anti-allergic activity (IC(50) = 18.7 ± 6.7 µM) in comparison with ketotifen fumarate (IC(50) = 15.0 ± 1.3 µM) and stronger anti-allergic activity than azelastine (IC(50) = 32.0 µM). Also, the MTT cytotoxicity assay with RBL-2H3 cells showed that δ-lactam does not display cytotoxicity at concentrations lower than 50 µM. This study suggests that the δ-lactam derivative has the potential to be used as a lead compound in the development of anti-allergic drugs for clinical use in humans.

West Nile Virus in the New World: trends in the spread and proliferation of West Nile Virus in the Western Hemisphere.

The observed patterns and variations in the ecology, epidemiology, distribution and prevalence of the West Nile Virus (WNV) in different areas of the Western Hemisphere make this pathogen of particular importance as a model for understanding the potential risk factors associated with emerging pathogens worldwide, particularly those involving zoonotic pathogens whose epidemiology involves the potential for vertical transmission in arthropod vector species, and horizontal and vertical transmission within and among vertebrate host species. Record numbers of human WNV cases were recorded in Canada during 2007, with >50% more cases than documented in any previous year. Although overall numbers of human infections recorded in the United States were not exceptionally high during 2007 relative to epidemic levels reported in 2002 and 2003, the state of Oklahoma reported that the highest-ever number of human WNV cases and the numbers of human cases recorded in Canada were 50% higher than previous record levels recorded in 2003. The record and near-record numbers of human WNV infections recorded in several regions of North America during 2007 have important implications for the future management and surveillance of WNV vectors and reservoirs in North America. The spatiotemporal distribution of WNV infections in humans and animals recorded during 2007 in North America and South America have important implications for the surveillance and management of public health threats from WNV in the Western Hemisphere. Serological surveys conducted in areas of intense WNV transmission in the United States have reported low prevalence of antibodies to WNV in human s populations, indicating that additional epidemic outbreaks of human disease from WNV can be expected in the future.

Human Dengue antibodies against structural and nonstructural proteins.

Antibodies against dengue virus type 2 and 4 proteins in acute-phase sera of 10 primary and 10 secondary dengue fever and dengue hemorrhagic fever patients were studied by Western blotting. In the first group the immune response was barely detectable, while in the second group more proteins were detected, with a very strong reaction. Anti-NS1 and -NS3 antibodies were detected mainly in secondary cases. Anti-E, -NS3, and -NS5 antibodies were detected in a high number of cases. The possibility of implementing early diagnostic assays for antigen detection is suggested.