488 resultados para PSYCHROTROPHIC CLOSTRIDIUM
Resumo:
Polyphenolic compounds occurring naturally in knotwood of plants are known to have antimicrobial effects. The knots (i.e. the branch bases inside tree stems) and outer branches in pine trees contain a remarkably high concentration of phenolic stilbenes, while lignans are the major phenolic constituents of spruce knots. Large amount of these phenolic compounds can be extracted from wood knots at pulp and paper mills where their presence is undesirable. In Finland, marinating of broiler meat is done not only to increase or add value to the meat, but also to enhance the safety and shelf-life. These products are usually packed under a modified atmosphere for further protection against spoilage microorganisms. However, studies have revealed that addition of marinades to poultry products do not have an inhibitory effect on either some psychrotrophic anaerobic bacteria, such as Brochothrix thermosphacta or lactic acid bacteria associated with spoilage. Also, the activity of pathogenic Campylobacter jejuni is not affected by marinating. The objective of this study was to investigate the inhibitory and lethal activities of extracts from spruce (Picea spp.) and pine (Pinus spp.) knotwood and outer branches that are dissolved in ethanol against the spoilage microorganisms in modified atmosphere packaged marinated broiler products. Modified atmosphere packaged broiler products were separately inoculated with ‘normal’ marinades, marinades with 70% ethanol, marinades with a mixture of spruce and pine extracts dissolved in 70% ethanol or mixture of spruce and pine extracts in powder form. The bacterial colony forming units per gram obtained from each of the samples were analysed on de Man Rogosa and Sharpe agar at days 1, 6, 12 and 15. The results showed that there were significant differences in bacterial colony forming units per gram (P <0.05) between packages with ‘normal’ marinades and packages with extracts added to their marinades on the 12th and 15th day. It can be concluded that the addition of extracts from spruce and pine knotwood to marinades significantly retarded growth of spoilage microorganisms during the 15 day test period. However further research is warranted to characterise and establish the safety and suitability of the compound(s) in spruce and pine knotwood extracts that are responsible for inhibitory or lethal activity against the microbes that may be present in marinated poultry meat.
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In cells, N-10-formyltetrahydrofolate (N-10-fTHF) is required for formylation of eubacterial/organellar initiator tRNA and purine nucleotide biosynthesis. Biosynthesis of N-10-fTHF is catalyzed by 5,10-methylene-tetrahydrofolate dehydrogenase/cyclohydrolase (FolD) and/or 10-formyltetrahydrofolate synthetase (Fhs). All eubacteria possess FolD, but some possess both FolD and Fhs. However, the reasons for possessing Fhs in addition to FolD have remained unclear. We used Escherichia coli, which naturally lacks fhs, as our model. We show that in E. coli, the essential function of folD could be replaced by Clostridium perfringens fhs when it was provided on a medium-copy-number plasmid or integrated as a single-copy gene in the chromosome. The fhs-supported folD deletion (Delta folD) strains grow well in a complex medium. However, these strains require purines and glycine as supplements for growth in M9 minimal medium. The in vivo levels of N-10-fTHF in the Delta folD strain (supported by plasmid-borne fhs) were limiting despite the high capacity of the available Fhs to synthesize N-10-fTHF in vitro. Auxotrophy for purines could be alleviated by supplementing formate to the medium, and that for glycine was alleviated by engineering THF import into the cells. The Delta folD strain (harboring fhs on the chromosome) showed a high NADP(+)-to-NADPH ratio and hypersensitivity to trimethoprim. The presence of fhs in E. coli was disadvantageous for its aerobic growth. However, under hypoxia, E. coli strains harboring fhs outcompeted those lacking it. The computational analysis revealed a predominant natural occurrence of fhs in anaerobic and facultative anaerobic bacteria.
Resumo:
Methylenetetrahydrofolate dehydrogenase-cyclohydrolase (FolD) catalyzes interconversion of 5,10-methylene-tetrahydrofolate and 10-formyl-tetrahydrofolate in the one-carbon metabolic pathway. In some organisms, the essential requirement of 10-formyl-tetrahydrofolate may also be fulfilled by formyltetrahydrofolate synthetase (Fhs). Recently, we developed an Escherichia coli strain in which the folD gene was deleted in the presence of Clostridium perfringens fhs (E. coli Delta folD/p-fhs) and used it to purify FolD mutants (free from the host-encoded FolD) and determine their biological activities. Mutations in the key residues of E. coli FolD, as identified from three-dimensional structures (D121A, Q98K, K54S, Y50S, and R191E), and a genetic screen (G122D and C58Y) were generated, and the mutant proteins were purified to determine their kinetic constants. Except for the R191E and K54S mutants, others were highly compromised in terms of both dehydrogenase and cyclohydrolase activities. While the R191E mutant showed high cyclohydrolase activity, it retained only a residual dehydrogenase activity. On the other hand, the K54S mutant lacked the cyclohydrolase activity but possessed high dehydrogenase activity. The D121A and G122D (in a loop between two helices) mutants were highly compromised in terms of both dehydrogenase and cyclohydrolase activities. In vivo and in vitro characterization of wild-type and mutant (R191E, G122D, D121A, Q98K, C58Y, K54S, and Y50S) FolD together with three-dimensional modeling has allowed us to develop a better understanding of the mechanism for substrate binding and catalysis by E. coli FolD.
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Changes in the quality of canned tilapia packed in oil and tomato sauce at ambient and accelerated temperatures were examined by microbiological and sensory evaluation. Canned tilapia were found to be microbiologically stable and organoleptically acceptable after six months storage period. Total viable count (TVC) were generally low (2.5 x 10 super(2)). Thermophilic organisms (Clostridium) were absent in all samples. The yield of edible part of tilapia was 72% after dressing. Pre-cooking of tilapia resulted in a loss of 21.5% of its dressed weight. Comparison of canned tilapia with available canned fishes (geisha and bonga) showed similar trends in the taste, proximate composition, microbiological stability and sensory scores.The possibility for investment in tilapia cannary was also investigated. It was found that production of canned tilapia will be economically viable if a ten hectare tilapia farm is used as a source of raw materials.
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Antagonistic activity of lactic acid bacteria (LAB) namely Streptococcus faecalis, Pediococcus cerevisiae and Lactobacillus casei was tested against seafood-borne bacteria such as Staphylococcus aureus, Bacillus cereus, Escherichia coli, Clostridium perfringens and Listeria monocytogenes. Three lactic acid bacteria such as Streptococcus faecalis, Lactobacillus casei and Pediococcus cerevisiae were coated on cooked mackerel meat, individually and in combination against fish-borne bacteria. S. faecalis inhibited C. perfringens in individual coat by 3.7 log units as compared to control, whereas L. casei did not inhibit C. perfringens. P. cerevisiae inhibited S. aureus by 5 log units. L. casei, inhibited L. monocytogenes by 3.3 log units on the third day of storage as compared to control. On the other hand, S. aureus and B. cereus were inhibited on the third and second day by 4.9 log and 5.2 log units respectively. B. cereus, S. aureus, L. monocytogenes were the most sensitive to all three LAB. C. perfringens was the least inhibited among all the seafood-borne bacteria tried. Multiple LAB or LAB strains in combination showed much earlier inhibitory activity on seafood-borne bacteria than single LAB coat.
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The results of the study carried out on the quality of fish preparations served in catering establishments of Bombay revealed that there is no serious potential health hazard to the consumer. Pathogens like Salmonella and Clostridium per were found to be absent. Based on organoleptic, biochemical and bacteriological parameters the quality of fish curry was better than that of fish fry. Overall quality of samples from grade I establishments was better in comparison with grade II and III. However, a few samples of poor quality were also observed in grade I. Extraneous matter like hair and dead housefly were observed in a few samples from grade III indicating poor handling practices. The importance of good hygiene and sanitary practices in catering establishments is discussed.
Resumo:
Study carried out on the quality of fresh fish in retail markets of Bombay revealed that only 75% of the samples were of acceptable quality. Incidence of faecal streptococci was generally high, indicating poor sanitary and hygienic practices in handling of fresh fish. Total bacterial counts higher than Indian standard specified limits were observed in more than one third of the samples analysed. 7.5% of the samples were found to be contaminated either with Salmonella or Clostridium perfringens, thus posing a serious potential health hazard to the consumer. The quality of fish in different markets is also discussed. The urgent need for formulation and implementation of quality standards for fresh fish in domestic trade is highlighted.
Resumo:
An examination was made of the rate of penetration of heat into fish sausage during processing at 115.6°C. Findings showed processing for 24 minutes to bring about complete destruction of Clostridium botulinum. A processing time of 30 minutes destroys almost all spoilage-causing organisms, thus prolonging the shelf life of the products.
Resumo:
Nisin is a widely used naturally occurring antimicrobial effective against many pathogenic and spoilage microorganisms. It has been proposed that reduced efficacy of nisin in foods can be improved by technologies such as encapsulation to protect it from interferences by food matrix components. The aim of this study was using of spray dried encapsulated nisin with zein in concentration of (0.15 and 0.25 g/kg) and sodium citrate (1.5 and 2.5%) and treatments with both of them to extent the shelf life of filleted trouts packaged by Modified Atmosphere Packaging (45% CO2, 50% N2 ,5% O2) and stored at 4±1 °C for 20 days. Furthermore, to evaluate the antimicrobial efficiency of encapsulated nisin and soudium citrate the trouts fillets was inoculated with Staphylococcus aureus as an index pathogenic bacteria. Assessment of chemical spoilage indexes such as (Proxide value, Thiobarbituric acid, total volatile base nitrogen and pH) , microbial parameters (Total Plate Count, Psychrotrophic count, Lactic acid bacteria count), Staphylococcus aureus cont in treatments which were inoculated with 5 logcfu/g of this bacteria and sensory evaluation of fillets including (smell, color, texture and total acceptability) was carried out in days of 0, 4, 8, 12, 16 and 20. The results revealed that treatment with both exposure of nisin and sodium citrate showed significantly lower chemical spoilage indexes in comparison with controls (vaccum packed and MAP) (P<0.05). Furthermore, (nisin 0.25 g/kg sodium citrate 2.5%) treatment which was exposed to the maximal level used of both materials was significantly the lowest treatment with (Proxide value, Thiobarbituric acid, total volatile base nitrogen and pH) of 9.95 (meq O2/kg) , 1.55 (mgMA/kg), 29.65 (mgN/100g) and 6.65 , respectively and according to the maximal recommended level of this indices , shelf life of fillets in this treatment was esstimated 20 days.The control (vaccum packed) treatment was significantly the highest treatment with (Proxide value, Thiobarbituric acid, total volatile base nitrogen and pH) of 15.17 (meq O2/kg), 3.03 (mgMA/kg), 38.4 (mgN/100g) and 6.95 , respectively and according to the maximal recommended level of this indices , shelf life of fillets in this treatment was estimated 11 days. Also, in microbial point of view (nisin 0.25 g/kg- sodium citrate 2.5%) treatment was the lowest treatment with Total Plate Count, Psychrotrophic count, Lactic acid bacteria count and Staphylococcus aureus count of 6.7, 6.83, 5.25 and 6.04 logcfu/g respectively, and conrol (vaccum packed) treatment was the highest treatment with 9.15, 9.41, 7.7 and 9.01 logcfu/g respectively. According to the lower results of chemical and microbial indices and higher sensory evaluated scores assessed in this research for encapsulated nisin in comparison with free nisin , it was concluded that encapsulation of nisin with zein capsules may improve the efficiency of nisin. The measuremented values of Mass yield, Total solids content of capsules, Encapsulation efficiency, In vitro release kinetics in 200 hour for encapsulated nisin in this study was 49.89, 62, 98.31 and 69% respectively and Encapsulated particle size was lower than 674.21 μm for 90% of particles. As a consequence, nisin , in particular encapsulated nisin, and sodium citrate alone or together with and Modified Atmosphere packaging might be considered as effective tools in preventing the quality degradation of the fillets, resulting in an extension of their shelf life.
Resumo:
Effects of different thawing method i.e. in a refrigerator, in water, at air ambient temperature and in a microwave oven on proximate, chemical (PV, TBA, FFA, TVB-N, SSP, FA), biochemical (pH, WHC,ThL), microbial (total viable, psychrotrophic, coliform, Shewanella and yeast-mould count) and sensory analysis were carried out on frozen whole Caspian sea Kutum (Rutilus frisii kutum) and Rainbow trout (Oncorhynchus mykiss) carcasses. The values of ash, protein, SSP, WHC, PUFA, PUFA/SFA. EPA+DHA/C16:0, pH, and microbial count of thawed samples decreased significantly while fat, PV, TBA, FFA, TVB-N, SFA and MUFA increased compared to the fresh fish (unfrozen) as control samples. Also, sensory evaluation all of thawed samples showed a significant (p<0.05) quality loss compared to the fresh fish as control samples. The lowest chemical and biochemical values as well as microbial growth were determined in water thawed samples. Therefore, based on this study thawing in water is most suitable for frozen whole rainbow trout.
Resumo:
本文从不同厌氧生境中获得7组(C-2、Y-2、L-2 、NZ、H-3、CZ、L-3)具有纤维素降解能力的复合菌系。经过不断传代、淘汰纤维素降解能力降低的菌系,最后得到一组高效、传代稳定的厌氧纤维素分解复合菌系L-3。该菌系可使滤纸在42 h内溃烂,并能在分解纤维素的同时产氢气。对L-3复合菌系的产酶条件进行了研究,结果表明,在实验范围内该菌系的产酶最适条件为:pH 6.5,温度37 ℃,接种量5 %,最佳碳源为滤纸,最佳氮源为硫酸铵。第10天测得羧甲基纤维素酶(CMCase)、滤纸酶(FPA)、外切葡聚糖酶(C1)、β-葡聚糖苷酶(β-glucodase)的酶活分别为0.216 U/ml、0.101 U/ml、0.132 U/ml、0.002 U/ml,滤纸失重率70.6 %。发酵代谢产物乙醇和丁酸含量分别可达1378 mg/L 、2695 mg/L,发酵产生的气体中氢气含量最高可达70.2 %。DGGE结果表明该菌系主要由14种菌组成,其中有三株菌在发酵前后菌数发生了明显的变化,说明在以滤纸为底物的降解过程中,这三株菌起到了重要作用,对这三株菌进行了分子生物学鉴定,初步定为Clostridium phytofermentans、Clostridium cellulovorans、Desulfovibrio sp。 利用实验室分离得到的纤维素降解菌,最终配制出由10、X-1、X-13、ST-13、L-3组成的好氧-厌氧纤维素降解复合菌剂。以秸秆为发酵底物,菌剂接种量1%,利用复合菌剂预处理后的秸秆,发酵总产气量相对于对照提高了71.62%,甲烷含量最高可达70.08%。 A group of microbial consortia L-3 was isolated from the anaerobic fermentation residue of corn stalk, which could degrade cellulose and produce hydrogen. The CMCase, FPA, C1 and β-glucosidase activity of L-3 could reach to 0.216 U/ml, 0.101 U/ml, 0.132 U/ml and 0.002 U/ml, respectively. In the filter degrading process, the filter paper collapsed in the liquid culture within 42 h and the filter degrading rate could reach to 70.6% in the 13 days, meanwhile, hydrogen was determined and the highest hydrogen content was 70.2%. The optimum cellulase-degrading conditions were filter papaer as the carbon source, (NH4)2SO4 as the nitrogen source, 37 ℃ and pH 6.5 in this experiment. DGGE results showed that the microbial consortia L-3 mainly included 14 strains. The amount of 3 strains were changed during the fermentation. These strains were identified as Clostridium phytofermentans、Clostridium cellulovorans、Desulfovibrio sp by 16S rDNA sequence analysis. The cellulose- degrading microbial agent was composed by 10, X-1, X-13, ST-13, L-3 which were isolated in the laboratory. The straw pretreated by cellulose-degrading microbial agent was used to ferment, the total biogas production increased by 72% comparing to the control. The content of methane could reach to 70.08%。
Resumo:
本文从四川绵竹酒厂、成都市龙泉长安垃圾填埋场以及四川大学荷花池底的厌氧污泥中先后分离得到63株厌氧产氢菌,其中H-8、H-61、HC-10等16株产氢细菌产氢能力较高,HC-10的产氢能力最高,最大产氢量和最大产氢速率分别达到2840 ml H2/L培养基和25.39 mmol H2/g drycell·h,对HC-10进行生理生化鉴定和分子生物学鉴定,判定其为clostridium sp.,对HC-10的产氢条件进行了研究,结果表明,该菌的最适生长温度为35 ℃,最适生长初始pH为7,以葡萄糖为最佳碳源,以蛋白胨为最佳氮源,不利用无机氮源,其产氢发酵液相产物以乙醇和乙酸为主,其发酵类型属于乙醇型发酵。此外,以酒糟废液作为底物,进行了菌株HC-10的生物强化试验,研究表明,投加了HC-10的强化系统其产氢量比对照高出40.32%。 同时为了获得厌氧产氢菌的高效突变株,分别以产氢菌H-8和H-61为原始菌株进行微波诱变处理,对微波诱变参数进行了优化,考察了突变株的遗传稳定性、产氢特性及耐酸性。菌株H-8经过微波诱变得到5株高产氢突变株HW7、HW33、HW181、HW184、HW195,经多次传代表明HW195是稳定的高产突变株。突变株HW195具有较好的耐酸性,在pH值为2.8时仍能生长。通过间歇发酵实验,其最大产氢量和最大产氢速率分别达到2460 mL/L培养基和27.97 mmol H2/g drycell·h,比原始菌分别提高了50.75%和41.7%。菌株H-61经过微波诱变后选育得到的突变株HW-18,其最大产氢量和最大产氢速率分别达到2190 mL/L培养基和25.86 mmol H2/g drycell·h,比原始菌分别提高了23.03%和31.00%。 为了对比各种诱变方式对产氢菌产氢能力的影响,以厌氧产氢菌H-61为原始菌株,先后经亚硝基胍(NTG)、紫外(UV)诱变,选育得到1株高产突变株HCM-23。在葡萄糖浓度为10 g/L的条件下,其产氢量为3024 mL/L培养基,比原始菌株提高了69.89%;其最大产氢速率为33.19 mmol H2/g drycell·h,比原始菌株提高了68.14%。经过多次传代实验,稳定性良好。其发酵末端产物以乙醇和乙酸为主,属于典型乙醇型发酵。其最适产氢初始pH为6.5,最适生长温度为36 ℃,以蔗糖为最佳碳源。与原始菌株相比,突变株HCM-23的产氢特性发生了改变,如生长延滞期延长,可利用无机氮源等。 From anaerobic activated sludge, 16 strains of hydrogen producing bacteria were newly isolated. One of them named as HC-10 had the highest hydrogen producing capability, under the batch fermentative hydrogen production condition, the maximal hydrogen yield and hydrogen production rate was 2840 mL/L culture and 25.39 mmol H2/g drycell·h. It was identified as clostridium sp.HC-10 by 16S rDNA sequence analysis. Various parameters for hydrogen production, including substrates, initial pH and temperature, have been studied. The optimum condition for hydrogen producing of strain HC-10 were achieved as: initial pH 7.0, temperature 35 ℃, glucose as the favorite substrate, Moreover, using distiller's solubles wastewater as substrate, HC-10 strain was added in the biohydrogen producing system to research the bioaugmentation effection. The results showed that the hydrogen production of bioaugmentation system was 40.32% higher than the noaugmentation system. An anaerobic, hydrogen producing strain H-8 was irradiated by microwave to optimize the microwave mutagenesis condition, and to test the heredity, hydrogen-producing potential and aciduric of the mutants. An aciduric mutant named as HW195 with steady hydrogen-producing capability was obtained, which can grow at pH 2.8. Its capability of hydrogen production was tested in the batch culture experiments. The maximum hydrogen yield and hydrogen production rate was 2460 mL/L culture and 29.97 mmol H2/g drycell·h, which was 50.7% and 41.7% higher than those of the initial strain, respectively. When used the strain H-61 as original strain, a mutant named as HW18 was obtained. The maximum hydrogen yield and hydrogen production rate was 2190 mL/L culture and 25.86 mmol H2/g drycell·h, which was 23.03% and 31.00% higher than those of the initial strain, respectively. The results demonstrated that microwave mutagenesis could be used in the field of hydrogen producing microorganism. The hydrogen producing strain H-61 was used as an original strain which was induced by NTG and UV for increasing and the hydrogen production capability. One of the highest efficient H2-producing mutants was named as HCM-23 with its stable hydrogen production capability. which was tested in the batch culture experiments. With the condition of 10 g/L glucose, its cumulative hydrogen yield and hydrogen production rate was 3024 mL/L culture and 33.19 mmol H2/g drycell·h, 69.89%and 68.14% higher than that of the original strain, respectively. The terminal liquid product compositions showed that the mutant HCM-23 fermentation was ethanol type, while the original strain H-61 fermentation was butyric acid type. Varieties of parameters of hydrogen production fermentation were studied, including time, carbon source, nitrogen source, glucose concentration, glucose utilization, initial pH and incubation temperature had been studied, indicated the optimum condition of hydrogen production for the mutant HCM-23 as initial pH6.5, temperature 36 ℃, and the favorite substrate was sucrose. The hydrogen production characters of the mutant and the original strain were different, such as, the growth lag phase and the utilization of inorganic nitrogen source, etc. This work shows a good application potential of NTG-UV combined mutation in the biohydrogen production. And the hydrogen production mechanism and metabolic pathway should be explored furthermore.
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从成都市洪河垃圾场采样分离到三株嗜热纤维素分解细菌,均为严格厌氧的革兰氏阴性杆菌,有较高的纤维素降解活性,能直接转化为纤维素产酒精。其中TCl菌株为周生鞭毛运动,不产生芽孢,大小为0.9×2.4~5.4um,表明菌落为白色,边缘整齐的圆形,最适温度在55℃,最适PH大约7.0,发酵纤维素或纤维而糖产生乙酸、丁酸。该菌属于拟杆菌料(Bacteroidaceae),不能归入已有的任何一属,很可能是一个新的属。TC3和TC4为单端丛生鞭毛运动,大小为0.6×2.4~15um,形成卵圆形或圆形的端生芽孢,使菌体膨大。TC3的菌落为灰白色、透镜状,TC4的菌落为黄色,透镜状,TC4在液体培养基中产生非水溶性黄色素。最适温度均为55℃,最适PH值TC3为8.0,TC4为7.5。TC3、TC4属于梭菌属,与所有已报道的同类菌株均有显著差异,因此认为它们可能是梭菌属 (clostridium)中的新种。
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从砷污染土壤中富集砷抗性细菌,在厌氧环境中进行培养,观察其对砷的还原能力。结果表明:在21h之内,As(V)就被完全还原为As(Ⅲ);培养72h后,培养基中出现黄色沉淀,采用X射线衍射分析(XRD)和扫描电镜-能谱分析(SEM-EDS)技术对沉淀进行分析表明,沉淀主要是以3种晶型存在的硫化砷(AsS);培养150h后,大约有65%的As以上述沉淀的方式从溶液中移除。此外,本文还采用了构建16SrDNA文库的方式对该体系中的微生物种群进行分析,利用RFLP技术对16SrDNA片段进行分型,共得到72个操作单元类型(OTU),其中6个OTU占了库容的51%,从这6个OTU中各选取1个克隆进行测序,结果表明,富集到的砷还原细菌属于喜热菌属(Caloramator)、梭菌属(Clostridium)和杆菌属(Bacillus)。
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The complex, K2.5Na2NH4[Mo2O2S2(cit)(2)]. 5H(2)O (1), was obtained by crystallization from a solution of (NH4)(2)MoS4, potassium citrate (K(3)cit) and hydroxyl sodium in methanol and water under an atmosphere of pure nitrogen at ambient temperature. The crystals are triclinic, space group
, a = 7.376 (3)Angstrom, b = 14.620 (2) Angstrom, c = 14.661 (1) Angstrom, alpha = 71.10 (1)degrees, beta = 81.77 (1)degrees, gamma = 78.27(2)degrees, R = 0.0584 for 2545 observed (I > 2 sigma (I)) reflections. Single crystal structure analysis reveals that citrate ligand coordinated to molybdenum atom through two carboxylato oxygens and one deprotonated hydroxyl oxygen together with two bridging sulfur atoms and a terminal oxygen atom completes distorted coordination octahedron around each molybdenum atom. Principal dimensions are: Mo = O-t, 1.707 Angstrom (av); Mo-S-b, 2.341 Angstrom (av); Mo-O-(hydroxyl), 2.021 Angstrom (av); Mo-O(alpha-carboxyl), 2.1290 Angstrom (av) and Mo-O(beta-carboxyl), 2.268(av) Angstrom. IR spectrum is in agreement with the structure.