Distribution of clinically important thermophilic actinomycetes in vegetable substrates and soil in north-western India

Medically important thermophilic actinomycetes were isolated from 218 (64%) of 341 samples of vegetable substrates and soil examined from sites in north-western India. Thermoactinomyces vulgaris (T. candidus) was the commonest species, occurring in 56% of samples, followed by Saccharomonospora viridis in 29%, Thermoactinomyces thalpophilus in 27%, Faenia rectivirgula (Micropolyspora faeni) in 21% and Thermoactinomyces sacchari in 14%. T. vulgaris and T. thalpophilus were isolated from all types of substrate examined, with T. vulgaris always more common than T. thalpophilus. Of the other thermophilic actinomycetes, F. rectivirgula was isolated predominantly from hay (44%) and S. viridis (56%) and T. sacchari (44%) from sugarcane bagasse. The largest populations of T. vulgaris and T. thalpophilus were found in paddy straw, followed by T. sacchari, S. viridis and F. rectivirgula in sugar-cane bagasse.The widespread occurrence of these clinically important thermophilic actinomycetes suggests that exposure of humans and animals to them may be frequent in north-western India. Studies are required to determine the prevalence of extrinsic allergic alveolitis (hypersensitivity pneumonitis) caused by thermophilic actinomycetes in the local population.     

Soil Contamination of Grass Biomass Hay: Measurements and Implications

A large area of unutilized or underutilized marginal land in the northeastern USA, some of which currently produce a mature hay crop for use as mulch, is potentially suitable for bioenergy crop production. Mature-mixed grass hay bales (n?=?1980) were sampled across New York in late summer and fall of 2011 and 2012 from 65 farms. Chemical analysis of 19 parameters, including gross calorific value, fiber, and elemental analyses, was conducted on the samples. In addition, 156 soil samples, representing 67 soil types, were analyzed for 12 to 14 parameters, using two different extraction procedures. Results indicate that hay composition is extremely variable among bales, and that much of the variation is due to soil contamination. Soil contamination reduced energy content of hay, a reduction best estimated from ash content of the hay. Standard plant analyses of contaminated hay samples determine total elemental content, but the same analyses only partially extract soil elements. A subset of samples showed that fiber analysis of soil-contaminated hay is problematic, with results impacted greatly by soil type and gravimetric filtration method. Aluminum, because of its low plant uptake potential, its high concentration in most soils, and its relatively moderate range in concentration across soil types compared to other soil elemental predictors, is the best indicator of soil contamination of biomass when the soil type is unknown. Evaluation of herbaceous plants for bioenergy parameters should include ash and Al analysis to assess soil contamination, which could significantly bias other compositional analyses.     

Improved medium for recovery and enumeration of the farmer's lung organism, Saccharomonospora viridis.

A new medium, which we propose to call R8, was developed for the isolation and enumeration of the thermophilic actinomycete, Saccharomonospora viridis. This organism has been implicated in a range of hypersensitivity pneumonitides, including farmer's lung, but is generally isolated in small numbers from contaminated environments. Recovery of S. viridis from moldy hay and mushroom compost on R8 medium was compared with recovery on conventional media. S. viridis was isolated from both substrates but in highest numbers and most consistently on the R8 medium. The selectivity of this medium was best observed when the sedimentation chamber method was used for hay samples. Here S. viridis accounted for up to 80% of the total number of actinomycetes recovered on R8 and could not be recovered on rifampin selective medium under the same conditions. R8 was also found to be an efficient recovery medium for a range of thermophilic actinomycetes from mushroom compost and for another allergenic species, Faenia rectivirgula, from moldy hay. Contamination of isolation plates by thermophilic bacilli was reduced on R8 compared with the activity on half-strength tryptone soy agar, supplemented with 0.2% casein hydrolysate, and this, together with specific improvements in S. viridis growth, accounts for the selective effect. It is possible that the occurrence of S. viridis and its role as a causative agent of hypersensitivity pnuemonitis have been underestimated by the use of suboptimal recovery protocols. It is hoped that use of R8 in conjunction with dilution plate techniques will generate information on the ecology of S. viridis and contribute to health risk assessment studies.     

Improved medium for recovery and enumeration of the farmer's lung organism, Saccharomonospora viridis

A new medium, which we propose to call R8, was developed for the isolation and enumeration of the thermophilic actinomycete, Saccharomonospora viridis. This organism has been implicated in a range of hypersensitivity pneumonitides, including farmer's lung, but is generally isolated in small numbers from contaminated environments. Recovery of S. viridis from moldy hay and mushroom compost on R8 medium was compared with recovery on conventional media. S. viridis was isolated from both substrates but in highest numbers and most consistently on the R8 medium. The selectivity of this medium was best observed when the sedimentation chamber method was used for hay samples. Here S. viridis accounted for up to 80% of the total number of actinomycetes recovered on R8 and could not be recovered on rifampin selective medium under the same conditions. R8 was also found to be an efficient recovery medium for a range of thermophilic actinomycetes from mushroom compost and for another allergenic species, Faenia rectivirgula, from moldy hay. Contamination of isolation plates by thermophilic bacilli was reduced on R8 compared with the activity on half-strength tryptone soy agar, supplemented with 0.2% casein hydrolysate, and this, together with specific improvements in S. viridis growth, accounts for the selective effect. It is possible that the occurrence of S. viridis and its role as a causative agent of hypersensitivity pnuemonitis have been underestimated by the use of suboptimal recovery protocols. It is hoped that use of R8 in conjunction with dilution plate techniques will generate information on the ecology of S. viridis and contribute to health risk assessment studies.     

Occurrence of thermophilic actinomycetes in natural substrates in Nigeria

Thermophilic actinomycetes were isolated from 163 (48.95%) of 333 samples of vegetable substrates and soil from different sites in Anambra and Enugu States in Nigeria.Thermoactinomyces (Tha.) vulgaris was the most common, occurring in 32.4% of samples whileTha. thalpophilus was isolated from 20.1%.Tha. sacchari, Saccharomonospora (Sam.) viridis andSaccharopolyspora (Sap.) rectivirgula were isolated from 3–10.5% of the samples examined.Streptomyces (Stm.) thermovulgaris occurred in 2.7% andThermomonospora spp. in 0.6% of samples. Except forStm. thermovulgaris and, perhaps,Thermomonospora spp., all the species isolated are known to be allergenic in man. The five known allergenic species were abundant in hay and maize silage.T. vulgaris andT. thalpophilus were also frequently isolated from rice straw.Tha. vulgaris, Tha. thalpophilus andSap. rectivirgula were all most numerous in hay whileTha. sacchari was most numerous in maize silage. The widespread occurrence of thermophilic actinomycetes in Nigeria emphasizes the need for investigation of their role in allergic respiratory disorders.     

Detecting the Seeds of Nassella neesiana in Large Round Hay Bales,by Means of Non-Destructive Core Sampling

In the last three decades or so there has been a significant increase in fodder trading, both in terms of the quantity of fodder traded and in terms of its economic value to the industry. Often, this fodder type may be supplied free of charge to graziers in distress due to circumstances that follow natural disasters such as bushfires, drought, and flood. However, because of the obvious urgency arising from these situations, it is suspected that much relief fodder may unintentionally pose an elevated risk for dispersal of weeds since it may be supplied from pasture not normally used for trade in fodder, and therefore is of unknown quality. Previous destructive method to detect weed propagules in bales of fodder are cumbersome, time consuming and of limited ecological and statistical value. Therefore, objective of this paper was to development of a convenient method to assess round pasture hay bales for the presence of weed propagules, to prevent unintentional spread of noxious species in hay bales. To examine this objective known quantity of seeds were added in a series of distributions to bales of seed free pasture hay, and a positive correlation for the amount of seed added per bale with that recovered in core samples was observed. Whilst the number of seeds detected per bale varied according to the distribution of seeds within the bales and the number of cores analysed, the absolute detection of seeds suggests that this sampling method is worthy of further examination. In addition, a pragmatic estimation of bale remnants after stock feeding has been investigated to more closely estimate the potential size of the remaining seed bank. The authors propose that development of this approach is timely, in the light of future climatic uncertainty driving extreme weather events that increase the need for relief fodder, which can be a potential vector for the spread of noxious weed seeds.     

Winter feeding sites of hay in round bales as major developmental sites of Stomoxys calcitrans (Diptera: Muscidae) in pastures in spring and summer

The stable fly, Stomoxys calcitrans (L.), historically has been a pest of livestock in confined operations but seldom of animals on pastures or rangelands. In the past two decades, however, S. calcitrans has become a major pest of cattle and horses on pastures in the midwestern United States. Although there usually is an overabundance of diverse stable fly and house fly, Musca domestica L., larval habitats in confined livestock operations, no larval habitat for stable flies has been clearly identified in the pasture-range environment. Because the winter feeding of hay in round bales results in significant amounts of hay wastage that when mixed with manure, might develop into suitable larval habitats, this study evaluated these areas as developmental sites for the abundant stable flies in pastures. There was a trend for fly traps placed in the vicinity of hay feeding sites to catch more stable flies than those placed distant from these sites. Estimates of stable flies emerging from these sites ranged from 102 to 1225 flies per core sample (25 by 25 cm). The mean number of adult stable flies during May and June 2001 through 2004 correlated negatively with the average minimum temperatures during the preceding winter (November-February) but not with rainfall or temperatures during the spring. These results support the hypothesis that winter feeding sites of hay in round bales are the main source of stable flies in pastures.     

Livestock Protection Dogs for Deterring Deer From Cattle and Feed

Abstract: Disease transmission between wildlife and livestock is a worldwide issue. Society needs better methods to prevent interspecies transmission to reduce disease risks. Producers have successfully used livestock protection dogs (LPDs) for thousands of years to reduce predation. We theorized that LPDs raised and bonded with cattle could be used to also reduce risk of bovine tuberculosis (Myobacterium bovis; TB) transmission between white-tailed deer (Odocoileus virginianus) and cattle by minimizing contact between the 2 species and use of cattle feed by deer. We evaluated 4 LPDs over 5 months, utilizing 2 data collection methods (direct observation and motion-activated video) on deer farms that supported higher densities than wild populations. Dogs were highly effective in preventing deer from using concentrated cattle feed (hay bales), likely the greatest risk factor of TB transmission on farms. Dogs also prevented deer from approaching cattle in core areas of pastures (near hay bales) and were very effective throughout pastures. Our research supports the theory that LPDs, specifically trained to remain with cattle, may be a practical tool to minimize potential for livestock to contract TB from infected deer in small-scale cattle operations. Where disease is present in deer, it may be possible to reduce the potential for disease transmission by employing LPDs.     

Effects of a Bacterial Hay Preservative (Pediococcus pentosaceus) on Hay under Experimental Storage Conditions

The abundant growth of molds and thermophilic actinomycetes in stored hay decreases its quality and can be hazardous for the producer who inhales these contaminants when the moldy hay is fed in closed barns. These microbes are responsible for a respiratory disease called farmer's lung. Products, including bacterial cultures that can be inoculated in hay, are available to prevent hay deterioration by molds and bacteria. The aim of this study was to verify the effectiveness of Pediococcus pentosaceus (a bacterial inoculant) in preventing hay deterioration at different humidity levels in a laboratory experiment. Mixtures of grasses (mostly alfalfa, timothy, and clover) placed in plastic bags were treated with the commercially available product (live culture of P. pentosaceus) at 500,000 and 5,000,000 CFU/g of hay and humidified at different levels (20, 25, 30, and 35%). Control batches of hay (untreated) were prepared at the same humidity levels. The growth of inoculated bacteria in hay, pH level, and hay deterioration were evaluated. Under these experimental conditions, the growth of P. pentosaceus was abundant only when it was inoculated in very moist hay (35% moisture), resulting in bacterium levels of 6.3 x 10(sup8) CFU/g after 30 days. This abundant growth did not prevent the pH from increasing (final pH of about 9.0), nor did it prevent molding. At lower humidity levels (20, 25, and 30%), the bacterial inoculant used did not grow and did not prevent hay deterioration.     

Circulating antibodies against thermophilic actinomycetes in farmers and mushroom workers

Sera from individuals exposed to moldy hay, mushroom compost and unexposed individuals, were studied for antibodies against thermophilic actinomycetes and Aspergillus fumigatus. Antigens extracted from Thermoactinomyces vulgaris strains isolated from China were used in the study along with antigens from standard strains to screen the sera of individuals exposed to farms and unexposed controls from both China and the United States. It was found that very little antibody response to A. fumigatus and F. rectivirgula was demonstrated in the sera of both individuals exposed to farm environments and unexposed controls. On the other hand, antibodies to Thermoactinomyces vulgaris and T. candidus strains were seen in the sera of both exposed and unexposed individuals from China, indicating that these organisms are prevalent in all the environments of China. The results also indicate that antigenicity of the strains vary considerably in their reactivity with sera.     

16S rRNA基因高通量测序方法检测奶牛场常用干草表面微生物群落结构及多样性

【目的】随着中国奶牛业的发展,干草需求量与日俱增。作为天然牧草,干草可以成为家畜传播病原体的载体。以干草表面附着物为研究对象,了解干草中细菌群落结构以及致病菌属特征。【方法】对来自6个不同奶牛场饲草舍的干草样本,应用Illumina Mi Seq高通量测序技术测定干草表面附着物细菌的16S r RNA基因V3-V4变异区序列,分析不同干草样本细菌群落组成。【结果】干草样本中的细菌在97%的相似水平下共得到OTU个数为15 416,涵盖了29门87纲144目219科323属的细菌。微生物多样性分析表明,干草样本具有很高的细菌多样性,不同样本多样性存在差异。对干草样本菌群中丰度较高的14种病原菌属进行分析,发现相较于人工种植牧草制备的干草,天然牧草制备的干草中病原菌属丰度较高。【结论】研究解析了干草样本中微生物的多样性、丰度及主要病原菌属的特征,对奶牛场疾病防控有一定指导意义。     

云南高原湖泊中高温放线菌四新种

在进行云南高原湖泊水生放线菌研究的过程中,分离到大量高温放线菌,按国内通用的方法进行鉴定,有4个新种。模式菌株Y86—8217,Y86—8100,Y86—8010,Y86—8048均保存于云南省微生物研究所。     

Fungal and actinomycete spore aerosols measured at different humidities with an aerodynamic particle sizer.

An aerodynamic particle sizer (APS) that uses laser Doppler velocimetry was used to determine aerodynamic diameters of spores of fungal and thermophilic actinomycete species common in mouldy hay, aerosolized at different humidities and temperatures. Results were compared with those obtained from inertial impaction in a cascade impactor. The APS gave slightly smaller measurements than the cascade impactor. Both methods gave aerodynamic diameters generally slightly smaller than the average spore dimensions observed on cascade impactor slides with a microscope. The latter measurements were less than axial dimensions given in the literature. Brief passage of spores through air at 95% relative humidity (RH) and 38 degrees C, compared with 40% RH and 20 degrees C, caused an immediate increase in their aerodynamic diameter and the breaking of chains of spores. Cultures maintained at 75% RH and aerosolized at 98% RH similarly produced larger spore particles than those passed through dry air. These findings have implications for mould-induced asthma and allergic alveolitis since they relate to physical behaviour of airborne spores and particle deposition sites in the lung.     

Coeliac disease with farmers' lung.

Two patients with allergic alveolitis due to mouldy hay antigens (farmer''s lung) were shown to have malabsorption due to coeliac disease. As similar associations have been found with other alveolar diseases, this association is probably not fortuitous and further population screening should be done.     

Fungi isolated from contaminated baled grass silage on farms in the Irish Midlands

The incidence of fungal growth on baled grass silage was recorded on 35 farms in the Irish Midlands in 2003. Fungal colonies were visible on 58 of 64 bales examined and the number of colonies per bale ranged from 1 to 12. On average, 5% of bale surface areas were affected. The fungus most prevalent on bales was Penicillium roqueforti, present on 86% of bales and representing 52% of all isolates. Other moulds isolated were Penicillium paneum, Geotrichum, Fusarium and mucoraceous species. Schizophyllum commune was observed protruding through the plastic film on bales on 17 of the 35 farms.     

Fungal and actinomycete spore aerosols measured at different humidities with an aerodynamic particle sizer

T.M. MADELIN AND H.E. JOHNSON. 1992. An aerodynamic particle sizer (APS) that uses laser Doppler velocimetry was used to determine aerodynamic diameters of spores of fungal and thermophilic actinomycete species common in mouldy hay, acrosolized at different humidities and temperatures. Results were compared with those obtained from inertial impaction in a cascade impactor. The APS gave slightly smaller measurements than the cascade impactor. Both methods gave aerodynamic diameters generally slightly smaller than the average spore dimensions observed on cascade impactor slides with a microscope. The latter measurements were less than axial dimensions given in the literature. Brief passage of spores through air at 95% relative humidity (RH) and 38°C, compared with 40% RH and 20°C, caused an immediate increase in their aerodynamic diameter and the breaking of chains of spores. Cultures maintained at 75% RH and aerosolized at 98% RH similarly produced larger spore particles than those passed through dry air. These findings have implications for mould-induced asthma and allergic alveolitis since they relate to physical behaviour of airborne spores and particle deposition sites in the lung.     

Microbial prevalence in domestic humidifiers.

The prevalence of viable thermophilic bacteria and actinomycetes and mesophilic fungi was examined in 145 samples from 110 domestic humidifiers. A total of 72 and 43% of furnace and console humidifier samples, respectively, contained viable thermophilic bacteria, whereas 60 and 72% of these samples produced mesophilic fungal growth. Thermophilic actinomycetes were recovered from seven humidifier samples. Efforts to detect thermophilic actinomycete antigens in 15 humidifier fluid samples were not successful. Antifoulants added to humidifier fluid reservoirs had no apparent effect on microbial frequency. Airborne microbial recoveries did not reflect patterns of humidifier contamination with respect to either kinds or numbers of microorganisms in 20 homes in which volumetric air samples were obtained during humidifier operation.     

Variation in microbial population during composting of agro-industrial waste

Two compost piles were prepared, using two ventilation systems: forced ventilation and ventilation through mechanical turning. The material to compost was a mixture of orange waste, olive pomace, and grass clippings (2:1:1 v/v). During the composting period (375 days), samples were periodically taken from both piles, and the enumeration of fungi, actinomycetes, and heterotrophic bacteria was carried out. All studied microorganisms were incubated at 25 and 55 °C after inoculation in appropriate growth media. Fungi were dominant in the early stages of both composting processes; heterotrophic bacteria proliferated mainly during the thermophilic stage, and actinomycetes were more abundant in the final stage of the composting process. Our results showed that the physical and chemical parameters: temperature, pH, moisture, and aeration influenced the variation of the microbial population along the composting process. This study demonstrated that composting of these types of wastes, despite the prolonged mesophilic stage, provided an expected microbial variation.     

Farmer’s Lung Disease and Microbiological Composition of Hay: A Case–Control Study

Previous studies performed in France have suggested that handling hay contaminated with high amounts of moulds, and especially Absidia corymbifera and Eurotium amstelodami, may favour farmer’s lung disease. The circumstances favouring farmer’s lung disease and the distinctive microbiological composition of hay samples that provoke attacks need to be specified. We present a case–control study which investigates the agricultural practices and the microbiological composition of hay handled in patients with farmer’s lung disease as compared to those of a representative control population. Ten cases identified the hay they were handling at the onset of symptoms. The location, type of farm and working conditions were similar to those of the control farms. Conversely, the microbiological composition of hay differed, with significantly higher amounts of E. amstelodami (P < 0.01), A. corymbifera (P = 0.003), mesophilic Streptomyces (P < 0.01), thermophilic Streptomyces (P < 0.01) and Saccharomonospora viridis (P < 0.01) than in the control population. Our results demonstrate that hay identified by patients as having a harmful effect is characterized by a higher total amount of microorganisms, notably five microorganisms that seem discriminative. Mean concentrations are 2- to 115-fold higher in hay suspected to cause symptoms than in hay from a representative panel of farms. Handling hay with high amounts of these five microorganisms constitutes a risk factor for farmer’s lung disease that should be considered for the development of prophylactic measures.     

Quarantine control of Hessian fly (Diptera: Cecidomyiidae) in exported hay: a new treatment for large-size, polypropylene fabric-wrapped bales and a 3-d fumigation for compressed standard bales

Hessian fly puparia (37,167), Mayetiola destructor (Say), did not survive a large-scale commercial test (three freight containers) of a new quarantine treatment using compression (32 kg/cm2) and hydrogen phosphide fumigation (61 g/28.2 m3) for large-size, polypropylene fabric-wrapped bales of hay exported to Japan. Mean +/- SEM temperatures in the large-size bales in different locations in the freight containers ranged from 18.0 +/- 0.9 to 26.0 +/- 1.3 degrees C during the 7-d test conducted in a heated building at 20.1 +/- 1.1 degrees C. Highest concentrations of hydrogen phosphide in most locations in the freight containers were observed after 3 d of fumigation and ranged from 366.7 +/- 96.1 to 425.0 +/- 162.7 ppm (mean +/- SEM) and throughout the 7 d of fumigation ranged from 253.6 +/- 59.9 to 407.1 +/- 76.5 ppm (mean +/- SEM). Hydrogen phosphide residues after fumigation and aeration were <10 ppb and were below the U.S. Environmental Protection Agency tolerance of 0.1 ppm in animal feeds. The results of the test fulfills regulatory agency testing requirements and confirms the efficacy of the treatment to control Hessian fly in large-size, polypropylene fabric-wrapped bales of hay. Hessian fly puparia (2,160) did not survive a large-scale commercial test of compression (105 kg/cm2) and a 3-d hydrogen phosphide (60 g/28.3 m3) fumigation for standard bales.