Bronchopneumonia with interstitial pneumonia in beef feedlot cattle: Characterization and laboratory investigation.

Bronchopneumonia with interstitial pneumonia (BIP) has been considered a variant of acute interstitial pneumonia (AIP) rather than a distinct disease. This study compared 18 BIP, 24 bronchopneumonia (BP), and 13 AIP cases in feedlot beef cattle. Grossly, BIP cases typically had cranioventral lung lesions of similar morphology and extent as BP cases, but the caudodorsal lung appeared overinflated, bulged on section, and had interlobular edema and emphysema. Gross diagnosis of BIP had 83% sensitivity and 73% specificity relative to histopathology. Histologic lesions of BIP in cranioventral areas were of chronic BP, while caudodorsal lesions included alveolar and bronchiolar damage and inflammation, interstitial hypercellularity, and multifocal hemorrhages. In BIP cases, cranioventral lung lesions were more chronic than caudodorsal lesions. Histologic scores and microbiology data were comparable in cranioventral lung of BIP versus BP cases and caudodorsal lung of BIP versus AIP cases, with differences reflecting a more chronic disease involving less virulent bacteria in BIP versus BP. Mycoplasma bovis infection was similarly frequent among groups, and a viral cause of BIP was not identified. Lesion morphology and similar blood cytokine concentrations among groups argued against sepsis as a cause of lung injury. Surfactant dysfunction was identified in BIP and BP, and was only partially the result of protein exudation. These and other findings establish BIP as a distinct condition in which chronic cranioventral BP precedes acute caudodorsal interstitial lung disease, supporting a role of chronic inflammation in heightened sensitivity to 3-methylindole or another lung toxicant.

Dung mimicry: the function of volatile emissions and corolla patterning in fly-pollinated Wurmbea flowers.

It has been suggested that flowers of some angiosperms mimic vertebrate faeces (dung) in order to exploit insect pollinators that utilize faeces as a source of food and/or oviposition sites. We investigated a potential case of mimicry in Wurmbea elatior (Colchicaceae), a lily that exhibits a faecal odour and pattern of dark spots on the corolla. We found that W. elatior is pollinated by a broad assemblage of coprophagous flies and is dependent on pollinator visits for seed production. The flowers emit volatiles that are characteristic of vertebrate faeces, and three of these compounds - skatole, indole, and an unidentified compound - elicited electrophysiological antennal responses from flies. Artificial flowers laced with indole and skatole or skatole alone attracted the same assemblage of flies as was recorded on flowers of W. elatior. Spotted artificial flowers attracted twice as many flies as did those lacking spots. Experimental addition of indole and skatole to flowers of Wurmbea kraussii, a congener with unscented flowers pollinated by hoverflies, induced a shift to an insect visitor assemblage dominated by coprophagous flies. This study clarifies the roles of volatile emissions (particularly skatole) and visual signals in floral dung mimicry.

An odorant receptor from Anopheles sinensis in China is sensitive to oviposition attractants.

BACKGROUND: Anopheles sinensis is an important vector for the spread of malaria in China. Olfactory-related behaviours, particularly oviposition site seeking, offer opportunities for disrupting the disease-transmission process. RESULTS: This is the first report of the identification and characterization of AsinOrco and AsinOR10 in An. sinensis. AsinOrco and AsinOR10 share 97.49% and 90.37% amino acid sequence identity, respectively, with related sequences in Anopheles gambiae. A functional analysis demonstrated that AsinOrco- and AsinOR10-coexpressing HEK293 cells were highly sensitive to 3-methylindole, but showed no significant differences in response to other test odorants when compared to DMSO. CONCLUSIONS: AsinOrco was characterized as a new member of the Orco ortholog subfamily. AsinOR10, which appears to be a member of the OR2-10 subfamily, is directly involved in identification of oviposition sites. This finding will help to elucidate the molecular mechanisms underlying olfactory signaling in An. sinensis and provide many more molecular targets for eco-friendly pest control.

Skatole remediation potential of Rhodopseudomonas palustris WKU-KDNS3 isolated from an animal waste lagoon.

UNLABELLED: Skatole (3MI) is a major contributor to the malodor emission resulting from ruminant and human faeces. The remediation of malodor has been a major challenge for the animal production industry. In this investigation, a pure culture of purple nonsulphur bacterium capable of degrading 3MI was isolated from a swine waste lagoon using an enrichment technique and identified as Rhodopseudomonas palustris WKU-KDNS3 based on 16S rRNA analysis and UV-visible spectroscopy. The cell structure of the organism was confirmed by transmission electron microscopy. Growth profile and 3MI removal pattern were determined using media supplemented with 0.1 µmol 3MI under short-term and long-term aerobic growth conditions. The organism grew on 3MI media as luxuriantly as control (without 3MI). Growth of R. palustris WKU-KDNS3 demonstrated a significant reduction in the level of 3MI (>48%) in 72 h. The level of 3MI dropped further by >93% of the total concentration present in the medium in 21 days. Skatole remediation potential of R. palustris WKU-KDNS3 can be judiciously utilized in various animal and industrial waste treatment systems. SIGNIFICANCE AND IMPACT OF THE STUDY: Odour pollution is a serious environmental problem, particularly in the agriculture industry, and technologies based on chemical remediation are less effective and cost prohibitive. In this study, the newly isolated Rhodopseudomonas palustris strain WKU-KDNS3 causes biodegradation of 3-methylindole (skatole), which is one of the most offensive odorants present in wastewater lagoons. Aerobic degradation of this widely spread aromatic pollutant by Rhodopseudomonas strain is a significant finding that enhances the present understanding about metabolic versatility of purple photosynthetic nonsulphur bacteria. The remediation potential of R. palustris WKU-KDNS3 can also be gainfully utilized in various waste treatment facilities.

A review on flavor of Baijiu and other world-renowned distilled liquors.

The flavor characteristics of distilled liquors significantly affect consumer acceptance and adoption. Therefore, odorants that contribute to sensory properties have received more attention. The odorants depend on the operating parameters, such as raw materials and ingredients, manufacturing process and maturing circumstances. This review summarized the odorants in the Baijiu and other world-renowned distilled liquors. Especially, the contribution of the odorants to the dominant aroma attributes is given more attention. The variations in the constituents and contents of odorants among the liquors are discussed comprehensively. In general, further research is still needed on the interaction mechanism between the odorants and sensory properties of distilled liquors.

Acute interstitial pneumonia and the biology of 3-methylindole in feedlot cattle.

Acute interstitial pneumonia (AIP) of cattle has been recognized for many decades. While the pathogenesis and risk factors for this condition in pastured cattle are relatively well characterized, there remains a poor understanding of the disease as it occurs in intensively fed cattle such as in beef feedlots. Specifically, in pastured cattle, AIP results from excessive ruminal production of the pneumotoxicant 3-methylindole (3-MI). In feedlot cattle, the evidence to substantiate the role of 3-MI is comparatively deficient and further investigations into the cause, pathogenesis, and control are sorely needed. This review highlights our current understanding of AIP with a focus on the disease as it occurs in feedlot cattle. Additionally, it illustrates the need for further work in understanding the specific animal factors (e.g. the ruminal microbiome, and the role of concurrent diseases), management factors (e.g. animal stocking and vaccination protocols), and dietary factors (e.g. dietary supplements) that may impact the development of AIP and which are relatively unique to the feedlot setting. All stakeholders in the beef industry stand to benefit from a greater understanding of what remains a pressing yet poorly understood issue in beef production.

Identification and functional characterization of olfactory indolergic receptors in Musca domestica.

In mosquitoes, indolic compounds are detected by a group of olfactory indolergic Odorant Receptors (indolORs). The ancient origin of indole and 3-methylindole as chemical signals suggest that they may be detected by insects outside the Culicidae clade. To test this hypothesis, we have identified potential indolOR genes in brachyceran flies based on sequence homology. Because of the crucial roles of indolic compounds in oviposition and foraging, we have focused our attention on the housefly Musca domestica. Using a heterologous expression system, we have identified indolOR transcript expression in the female antennae, and have characterized MdomOR30a and MdomOR49b as 3-methylindole and indole receptors, respectively. We have identified a set of 92 putative indolOR genes encoded in the genomes of Culicoidea, Psychodidae and brachycera, described their phylogenetic relationships, and exon/intron structures. Further characterization of indolORs will impact our understanding of insect chemical ecology and will provide targets for the development of novel odor-based tools that can be integrated into existing vector surveillance and control programs.

Identification and functional characterization of olfactory indolergic receptors in Drosophila melanogaster.

Indole-sensitive odorant receptors or indolORs belong to a mosquito-specific expansion as ancient as the Culicidae lineage. Brachyceran flies appeared to lack representative members of this group despite the importance of indolics in this important group of dipterans. To explore whether indolORs occur in other brachyceran species, we searched for candidate indolORs in Drosophila melanogaster. Using phylogenetic tools, we show that D. melanogaster OR30a, OR43a, and OR49b form a distinct monophyletic lineage with mosquito indolORs. To explore a potential functional orthology with indolORs, we expressed these three Drosophila ORs in Xenopus laevis oocytes and measured their responses to a panel of indolic compounds. We provide evidence that OR30a, OR43a, and OR49b exhibit high sensitivity to indoles. Along with the recent discovery of indolORs in the housefly Musca domestica, our findings suggest that indolORs are a widespread feature of the peripheral olfactory systems of Diptera.

[Experimental study on the effect of olfactory training on olfactory function in mice with olfactory dysfunction].

Objective: To observe the effect of olfactory training on mice with olfactory dysfunction induced by 3-methylindole (3-MI). Methods: Thirty-one male BALB/c mice were randomly divided into 3 groups by random digits table: control group (group A, n=10), olfactory dysfunction group (group B, n=10) and olfactory dysfunction+olfactory training group (group C, n=11). Mice in group B and group C were intraperitoneally injected with 150 mg/kg 3-MI to induce olfactory dysfunction model, while mice in group A were intraperitoneally injected with corn oil of the same volume. From the first day after injection, mice in group C were treated with 4 kinds of odors by inhalation, while mice in group B were treated with distilled water by inhalation, with 2 times/d, 30 min/time/kind of odor, and continuous training for 28 d. Group A was not treated. Buried food pellet tests were conducted before injection and at 7, 14, 21 and 28 days after injection, respectively. The olfactory epithelium was harvested for observation of the number of olfactory marker protein (OMP) and the thickness of olfactory epithelium on the 28th day after injection. SPSS 23.0 software was used for statistical analysis. Results: Before injection, all mice in each group had no olfactory dysfunction. At the 7th, 14th, 21st and 28th days after injection, the food finding time of mice in group C was shorter than that in group B, and the difference was statistically significant ((175.88±100.50) s vs (266.73±46.83) s, (132.00±84.62) s vs (264.10±48.50) s, (103.57±77.43) s vs (197.43±69.78) s, (67.79±32.54) s vs (176.63±61.06) s, all P<0.05), but food finding time of mice in group B and C was longer than that in group A (the food finding time of group A at the 7th, 14th, 21st and 28th days after injection was (27.13±5.36) s, (25.83±7.28) s, (23.13±2.72) s, (26.63±7.60) s, respectively, all P<0.05). At the 28th day after olfactory training, the number of OMP positive cells in group B and C were fewer than that in group A, and the difference was statistically significant ((108.00±28.19)/HP vs (288.22±84.06)/HP, (199.33±58.55)/HP vs (288.22±84.06)/HP, all P<0.05). The number of OMP positive cells in group C were higher than that in group B (P<0.05). The number of OMP positive cells had negative correlation with food finding time (r=-0.886, P<0.01). As for the thickness of the olfactory epithelium, the thickness of group B was thinner than that in group A and C, and the difference was statistically significant ((59.57±31.27) µm vs (114.55±40.70)µm vs (90.54±37.72) µm, all P<0.05). Conclusion: Olfactory training can accelerate the recovery of olfactory function in 3-MI-induced olfactory impaired mice.

Role of Mouse Cytochrome P450 Enzymes of the Cyp2abfgs Subfamilies in the Induction of Lung Inflammation by Cigarette Smoke Exposure.

Many constituents of tobacco smoke (TS) require bioactivation to exert toxic effects; however, few studies have examined the role of bioactivation enzymes in the adverse effects of TS exposure. This knowledge gap is a major source of uncertainty for risk assessment and chemoprevention efforts. Our aim is to test the hypothesis that cytochrome P450 (P450) enzyme-mediated bioactivation is essential to the development of TS exposure-induced lung toxicity, by determining the contributions of P450 enzymes in the mouse Cyp2abfgs gene subfamilies to environmental tobacco smoke (ETS)-induced lung inflammation. Adult female wildtype (WT) and Cyp2abfgs-null mice (both on C57BL/6J background) were exposed to filtered air or ETS, intermittently, for 1 or 2 weeks. Lung inflammation was assessed by quantification of inflammatory cells, cytokines, chemokines, and proteins in bronchoalveolar lavage fluid (BALF) and histopathological analysis. Glutathione (GSH) conjugates of 2 ETS constituents, naphthalene (NA), and 3-methylindole (3MI), were measured in mice exposed to ETS for 4 h. Persistent macrophagic and neutrophilic lung inflammation was observed in ETS-exposed WT mice; the extent of which was significantly reduced in ETS-exposed Cyp2abfgs-null mice. Levels of proinflammatory cytokines and chemokines, along with the total protein concentration, were increased in cell-free BALF from ETS-exposed WT mice, but not Cyp2abfgs-null mice. Additionally, GSH conjugates of NA and 3MI were detected in the lungs of WT, but not Cyp2abfgs-null, mice following ETS exposure. These results provide the first in vivo evidence that the mouse Cyp2abfgs gene cluster plays an important role in ETS-induced lung inflammation.

Characterization of key odorants causing the roasted and mud-like aromas in strong-aroma types of base Baijiu.

The roasted and mud-like aromas in Chinese strong-aroma types of base Baijiu (base SAB) and the interactions among the corresponding key compounds causing these two characteristic aromas were investigated. A total of 68 and 64 odorants were identified in the base and commercial SAB by comparative aroma extract dilution analysis. Furthermore, the complex matrix of Baijiu always pose a long-term challenge for quantitative accuracy and precision of target aroma compounds; thus, the odorants with flavor dilution (FD) ≥ 9 were further quantified by multiple quantitative techniques, including the direct injection combined with gas chromatography-flame ionization detector, the vortex-assisted surfactant-enhanced emulsification liquid-liquid microextraction (VSLLME) combined with GC-MS, and the derivatization method combined with VSLLME-GC-MS, and 37 and 26 odorants were shown to be important odorants as their OAVs ≥1. Among these components, 5 compounds, hexanoic acid, butyric acid, p-cresol, 3-methylindole, and 3-(methylthio)propanal showed higher OAVs in base than in commercial SAB, and based on aroma recombination and omission analysis, they were confirmed to be the key odorants responsible for the mud-like and roasted odors. The evaluation of the aforementioned five key odorants by the "addition" test showed that the roasted and mud-like aromas of base SAB were related, and the odors were mainly attributed to additive or masking effects among the compounds.

The effectiveness of platelet-rich plasma in an anosmia-induced mice model.

OBJECTIVE: In this study, we aimed to functionally and morphologically demonstrate the effectiveness of platelet-rich plasma (PRP) on anosmia in a mouse model of anosmia. STUDY DESIGN: Animal study. METHODS: A total of 16 male mice were included. When selecting the mice, the food-finding test (FFT) was used to make sure that the animals could smell, and anosmia was induced by administration of intraperitoneal 3-methylindole. The mice were randomly divided into two groups of eight (groups A and B). After 1 week, topical PRP was administered to the mice in group A and topical saline was administered to the mice in group B. The FFT was again administered at 7, 14, and 21 days. The mice were sacrificed on day 21, the olfactory neuroepithelium was histopathologically examined, and the epithelial damage scores and epithelial thickness were measured. RESULTS: After topical administration of PRP and saline, the difference in the average FFT values of the groups was statistically significant at 7, 14, and 21 days (P < 0,005). During the histopathological examination, the epithelial damage score was statistically significantly lower in the PRP group (P = 0.001) than in the saline group, and epithelial thickness was statistically significantly greater in the PRP group compared to the saline group (P = 0.003). CONCLUSION: We showed that PRP administration has a curative effect on olfactory functions in an anosmia-induced mice model. However, there is a need for further research before PRP can be considered for use in patients with anosmia in clinical practice. LEVEL OF EVIDENCE: NA. Laryngoscope, 128:E157-E162, 2018.

Characterization of 3-Methylindole as a Source of a "Mud"-like Off-Odor in Strong-Aroma Types of Base Baijiu.

The "mud"-like off-odor, which is an odor reminiscent of musty and strong animal fecal odors, affects the quality classification of the strong-aroma types of baijiu (SAB), but little is known about which compounds are responsible for this aroma, except for certain phenolic compounds. However, not all mud-like off-odor expressions in SAB can be attributed to those phenolic compounds. In this work, volatile compounds in pit mud and SAB samples were isolated by headspace solid-phase microextraction and liquid-liquid extraction. An odoriferous zone was detected by GC-MS/olfactometer and GC-MS and was attributed to 3-methylindole rather than ethyl oleate on the basis of their odor characteristics, MS data, and retention indices. Combined with the detection threshold of 3-methylindole determined by a three-alternative forced-choice (6.09 µg/L) procedure, odor activity values in the tested base SAB were from 1 to 23. Sensory studies and the prepared heat map highlighted the contribution of this compound to the odor characteristics for the first time.

Spectroscopic study of 2-methylindole and 3-methylindole: Solvents interactions and DFT studies.

The interaction between the 2- and 3-methylindole derivatives has been studied using spectroscopic technique and molecular modeling. Solvent polarity was found to play a great role in the interaction with acetonitrile showing greatest interaction with indole derivatives but less hydrogen bonding contribution. Good agreement was found between the models used for the analysis of experimental data and the theoretical calculations. Moreover, on the part of the quenching, benzonitrile was found to be better quencher over chlorobenzene, with 3-methylindole giving higher dynamic quenching constants.

Characterisation of aroma-active and off-odour compounds in German rainbow trout (Oncorhynchus mykiss). Part II: Case of fish meat and skin from earthen-ponds farming.

Odorous molecules in earthen-ponds rainbow trout aquaculture farming in Germany were investigated with a special focus on musty-earthy off-odorants. To this aim, fish meat and skin were extracted using solvent-assisted flavour evaporation (SAFE) and were mildly concentrated; extracts were subsequently analysed by means of one- and two-dimensional high-resolution gas chromatography coupled with mass spectrometry and olfactometry (GC-MS/O and 2D-HRGC-MS/O). Aroma extract dilution analysis (AEDA) of the solvent extracts revealed the presence of 76 odorants of which 75 were successfully identified. Thereby, rotundone (black pepper) is described for the first time as an odour-active substance in fish. Moreover, a series of compounds is described for the first time in German aquaculture rainbow trout fish, including, amongst others, (E,Z,Z)-2,4,7-tridecatrienal, (E)-4,5-epoxy-(E)-2-decenal, 4-ethyloctanoic acid, 3-methylindole (skatole), d-limonene, and indole. The analytical findings were further compared to sensory evaluation of the samples, and previously obtained data on the respective aquacultural water.

Phase I metabolism of 3-methylindole, an environmental pollutant, by hepatic microsomes from carp (Cyprinus carpio) and rainbow trout (Oncorhynchus mykiss).

We studied the in vitro metabolism of 3-methylindole (3MI) in hepatic microsomes from fish. Hepatic microsomes from juvenile and adult carp (Cyprinus carpio) and rainbow trout (Oncorhynchus mykiss) were included in the study. Incubation of 3MI with hepatic microsomes revealed the time-dependent formation of two major metabolites, 3-methyloxindole (3MOI) and indole-3-carbinol (I3C). The rate of 3MOI production was similar in both species at both ages. No differences in kinetic parameters were observed (p = 0.799 for Vmax, and p = 0.809 for Km). Production of I3C was detected only in the microsomes from rainbow trout. Km values were similar in juvenile and adult fish (p = 0.957); Vmax was higher in juvenile rainbow trout compared with adults (p = 0.044). In rainbow trout and carp, ellipticine reduced formation of 3MOI up to 53.2% and 81.9% and ketoconazole up to 65.8% and 91.3%, respectively. The formation of I3C was reduced by 53.7% and 51.5% in the presence of the inhibitors ellipticine and ketoconazole, respectively. These findings suggest that the CYP450 isoforms CYP1A and CYP3A are at least partly responsible for 3MI metabolism. In summary, 3MI is metabolised in fish liver to 3MOI and I3C by CYP450, and formation of these metabolites might be species-dependent.

Development and validation of a UHPLC-HR-Orbitrap-MS method for the simultaneous determination of androstenone, skatole and indole in porcine meat and meat products.

Boar taint is an off-odour that entails negative consumer reactions. In this study two extraction and UHPLC-HRMS analysis methods, valuable for evaluation of consumer acceptance towards boar meat, were developed for quantification of indole, skatole, and androstenone in different meat products. Sample pretreatment consisted of extraction with methanol and a homogenising step (cooked ham, minced meat, tenderloin, bacon, cutlets, blade loin, uncooked ham) or a melting step (salami sausage and liver paste). Both methods were validated according to CD 2002/657/EC and ISO 17025 guidelines. Good performance characteristics were obtained. Good linearity (R(2) ⩾ 0.99) and no lack of fit was observed (95% confidence interval; F-test, p > 0.05). Also good recovery (89-110%) and satisfactory precision: repeatability (RSD ⩽ 14.9%) and within-laboratory reproducibility (RSD ⩽ 17.2%) were obtained. Analysis of cooked ham and salami sausage samples proved the applicability of both methods for routine analysis.

Feedlot Acute Interstitial Pneumonia.

Acute interstitial pneumonia (AIP) of feedlot cattle is a sporadically occurring respiratory condition that is often fatal. Affected cattle have a sudden onset of labored breathing. There is no confirmed effective treatment of feedlot AIP; however, administration of antibiotics effective against common bacterial respiratory pathogens and nonsteroidal anti-inflammatory drugs, especially aspirin, has been recommended. Protective strategies are not well defined, but efforts to limit dust exposure and heat stress; to ensure consistent formulation, mixing, and delivery of feed; and to identify and treat infectious respiratory disease in a timely manner may decrease rates of feedlot AIP.

Complete genome sequence of a malodorant-producing acetogen, Clostridium scatologenes ATCC 25775(T).

Clostridium scatologenes ATCC 25775(T) is an acetogenic anaerobic bacteria known to be capable of synthesizing volatile fatty acids and solvents from CO2 or CO on its autotrophic mode and producing 3-methylindole and 4-methylphenol on its heterotrophic mode. Here, we report the complete genome sequence of this strain, which might provide a lot of valuable information for developing metabolic engineering strategies to produce biofuels or chemicals from greenhouse gases.

Effect of branched-chain fatty acids, 3-methylindole and 4-methylphenol on consumer sensory scores of grilled lamb meat.

Tenderness, flavour, overall liking and odour are important components of sheepmeat eating quality. Consumer assessment of these attributes has been made for carcasses from the Information Nucleus Flock (INF) of the Cooperative Research Centre for Sheep Industry Innovation. The concentrations of three branched chain fatty acids, 4-methyloctanoic (MOA), 4-ethyloctanoic (EOA) and 4-methylnonanoic acids (compounds related to 'mutton flavour' in cooked sheepmeat) and 3-methylindole and 4-methylphenol (compounds related to 'pastoral' flavour) were determined for 178 fat samples taken from INF carcasses. Statistical modelling revealed that both MOA and EOA impacted on the 'Like Smell' consumer sensory score of the cooked meat product (P<0.05), with increasing concentration causing lower consumer acceptance of the product. None of the compounds though had an effect on the liking of flavour. Obviously, reducing the effect of MOA and EOA on the odour of grilled lamb will improve consumer acceptance of the cooked product but other factors affecting the eating quality also need to be considered.