Biotechnological Methods to Accelerate Cheddar Cheese Ripening

ABSTRACT

Cheese is one of the dairy products that can result from the enzymatic coagulation of milk. The basic steps of the transformation of milk into cheese are coagulation, draining, and ripening. Ripening is the complex process required for the development of a cheese's flavor, texture and aroma. Proteolysis, lipolysis and glycolysis are the three main biochemical reactions that are responsible for the basic changes during the maturation period. As ripening is a relatively expensive process for the cheese industry, reducing maturation time without destroying the quality of the ripened cheese has economic and technological benefits. Elevated ripening temperatures, addition of enzymes, addition of cheese slurry, attenuated starters, adjunct cultures, genetically engineered starters and recombinant enzymes and microencapsulation of ripening enzymes are traditional and modern methods used to accelerate cheese ripening. In this context, an up to date review of Cheddar cheese ripening is presented.     

Milk microbiome signatures of subclinical mastitis-affected cattle analysed by shotgun sequencing

Aims: Metagenomic analysis of milk samples collected from Kankrej, Gir (Bos indicus) and crossbred (Bos taurus × B. indicus) cattle harbouring subclinical mastitis was carried out by next‐generation sequencing 454 GS‐FLX technology to elucidate the microbial community structure of cattle milk. Methods and Results: Milk samples from Kankrej, Gir and crossbred cattle were subjected to metagenomic profiling by pyrosequencing. The Metagenomic analysis produced 63·07, 11·09 and 7·87 million base pairs (Mb) of sequence data, assembled in 264 798, 56 114 and 36 762 sequences with an average read length of 238, 197 and 214 nucleotides in Kankrej, Gir and crossbred cattle, respectively. Phylogenetic and metabolic profiles by the web‐based tool MG‐RAST revealed that the members of Enterobacteriales were predominant in mastitic milk followed by Pseudomonadales, Bacillales and Lactobacillales. Around 56 different species with varying abundance were detected in the subclinically infected milk. Escherichia coli was found to be the most predominant species in Kankrej and Gir cattle followed by Pseudomonas aeruginosa, Pseudomonas mendocina, Shigella flexneri and Bacillus cereus. In crossbred cattle, Staphylococcus aureus followed by Klebsiella pneumoniae, Staphylococcus epidermidis and E. coli were detected in descending order. Metabolic profiling indicated fluoroquinolones, methicillin, copper, cobalt–zinc–cadmium as the groups of antibiotics and toxic compounds to which the organisms showed resistance. Sequences indicating potential of organisms exhibiting multidrug resistance against antibiotics and resistance to toxic compounds were also present. Interestingly, presence of bacteriophages against Staph. aureus, E. coli, Enterobacter and Yersinia species was also observed. Conclusions: The analysis identified potential infectious organisms in mastitis, resistance of organisms to antibiotics and chemical compounds and the natural resistance potential of dairy cows. Significance and Impact of the Study: The findings of this study may help in formulating strategies for the prevention and treatment of mastitis in dairy animals and consequently in reducing economic losses incurred because of it.     

Chemical Studies on Components of Dried Bonito, “Katsuobushi”

The flavor concentrate obtained by the extraction of “Katsuobushi” of bonito (Katsuwonus pelamis) with 80% ethanol and by the subsequent steam distillation of the extract was fractionated by the usual method and the resulting neutral, non-carbonyl oxygenated fraction was investigated by gas chromatography. The following components were tentatively identified: 2-pentanol and 2-methyl-1-heptanol as free alcohols, and 4 alcohols of n- and isobutanol, n-pentanol and n-dodecanol and 9 carboxylic acids of propanoic, n-butanoic, n-pentanoic, n-octanoic, n-nonanoic n-decanoic, n-dodecanoic, n-tetradecanoic and n-hexadecanoic acid as the constituents of esters. A constituent alcohol existing in the largest amount was isolated by gas chromatography and identified as 2-methyl-1-heptanol by elemental analysis, NMR, IR, and MS. A constituent acid existing in large amount was also isolated and investigated similarly, and the structure was partially estimated. 2-Methyl-1-heptanol holds a fresh woody aroma and seems to have a major effect on “Katsuobushi” flavor.     

Microbiological quality of raw milk produced in Estonia

Aims: The microbial quality of farm bulk‐tank raw milk produced in Estonia during years 2004–2007 was investigated. Methods and Results: Bulk‐tank milk samples were analysed for lactic acid bacteria count (LABC), psychrotrophic bacteria count (PBC), aerobic spore‐forming bacteria count (ASFBC), total bacterial counts using BactoScan and somatic cell count (SCC) using Fossomatic. Randomly selected psychrotrophic isolates were subjected to 16S–23S PCR‐ribotyping. LABC remained below 10⁴ CFU ml^?1 in most samples, while psychrotrophic micro‐organisms dominated in 60% of farms. PBC ranged from 4·2 × 10² to 6·4 × 10⁴ CFU ml^?1, and ASFBC varied from 5 to 836 CFU ml^?1. Conclusions: In general, the microbiological quality of the farm bulk‐tank milk was good – more than 91% of samples contained <50 000 CFU ml^?1, and SCC in the majority of samples did not exceed the internationally recommended limits. Genus Pseudomonas spp. was the dominating spoilage flora with Pseudomonas fluorescens as the prevailing species. Significance and Impact of the Study: Specific bacterial groups (LABC, PBC and ASFBC), not analysed routinely by dairies, were determined in bulk‐tank raw milk of numerous dairy farms during 4‐year period. Based on the survey, dairy plants can better control their supply chains and select farms (milk) for the production of specific products, i.e. milk with low PBC and high LABC for cheesemaking.     

Vergleich eines "glatten" und eines "rauhen" Stammes von Pseudomonas syringae pv. phaseolicola

Comparison of a “smooth” and a “rough” isolate of Pseudomonas syringae pv. phaseolicola The “smooth” (S) wild strain of Pseudomonas syringae pv. phaseolicola was compared with a “rough” (R) variant of low virulence. Both strains grew nearly equally well on a sucrose containing medium with yeast extract and casamino acids, and the strains did not differ markedly in the quantity of produced EPS (= extracellular polysaccharides). Principally the same results were obtained for high and medium concentrations of sucrose, or when sucrose was replaced by glucose or fructose. However, on glucose and fructose considerably lower quantities of EPS were produced. The biological activity of S-EPS was higher than that of R-EPS. This difference between the EPS preparations was not as marked as leaf inoculation with both bacterial isolates. After prolonged bacterial culture the EPS-production increased further, so that the differences between both strains decreased. A different EPS type was produced on the glycerol containing medium of KING B. Variations in the composition of this medium resulted in different morphology of the agar grown cultures, and the relative differences between S and R bacteria changed. When 62 different physiological tests for both bacterial strains were compared, the “rough” bacteria revealed a lowered range of positive reactions, with a few exceptions. However, it appeared unlikely that the reduced virulence of the “rough” bacteria was due to these differences. Obviously, defects in the extracellular products, but not in levan, were responsible for the reduction of virulence.     

Metal concentrations in certain coastal organisms from Beirut

Trace metal concentrations were determined in six species of intertidal organisms common to the coast of Ras Beirut, Lebanon. Lead, cadmium, nickel, iron, and zinc were highest in the polychaete,Hermodice carunculata, but the eggs of the sea urchin,Arbacia lixula, had similar iron levels and the sea anemone,Actinia equina, had zinc concentrations which also approached levels in the polychaete. The highest copper occurred in the shore crab,Pachygrapsus transversus, while chromium was highest in the eggs ofArbacia lixula.Cystoseira spinosa, the only alga studied, had average copper and iron concentrations similar to those found in the same species in another study. Together with the sea urchin eggs,C. spinosa exhibited the most variable zinc levels in the present investigation. The fish,Thalassoma pavo, which is rather common along the rocky coastal areas of Lebanon, appeared to have fairly high concentrations of certain elements in relation to levels reported in species of fish from other locations.Sewage, garbage, industrial and agricultural waste materials all enter the Mediterranean from Lebanon without prior treatment, which along with increased land erosion, probably contribute substantially to the availability of metals to the biota studied. It is suggested that more work on trace elements in coastal organisms from the eastern Mediterranean basin be undertaken before any conclusive statements are made. Such work should also investigate the various physiological and biochemical factors involved in metal uptake and retention by each species.     

Bleu cheese flavor production by submerged fermentation with on-line monitoring by gas chromatography

Eight cultures ofPenicillium roqueforti and related species were compared in shake-flask cultures on the basis of aroma.P. decumbens IFO 7091 was chosen as having the aroma closest to real Bleu cheese, and was used for fermentation studies. Gas chromatography was employed to obtain a continuous record of 2-pentanone concentration in the fermentor. Methyl ketones are characteristic of Bleu cheese flavor, and a subjective correlation was observed between the aroma of the fermentor off-gas and the 2-pentanone concentration measured. Cell growth and milk fat utilization increased consistently with increasing agitation and aeration, but 2-pentanone production did not correlate with growth and was maximum at intermediate values of agitation and aeration. The maximum 2-pentanone concentration, 100 ppm, was approximately ten times the concentration in Bleu cheese on a dry solids basis. Production of Bleu cheese flavor by submerged fermentation offers the opportunity for a new commercial value-added product from butterfat.Mention of brand or firm names does not constitute an endorsement by the US Department of Agriculture over others of a similar nature not mentioned     

Egg Trading in the Chalk Bass,Serranus tortugarum,a Simultaneous Hermaphrodite

The simultaneously hermaphroditic fish, Hypoplectrus nigricans, engages in a form of reciprocal spawning called “egg trading”, where one partner gives up eggs for fertilization in exchange for the opportunity to fertilize those of the partner. Since egg trading appears to be important in the evolutionary maintenance of hermaphroditism in H. nigricans, an attempt was made to determine if another species, Serranus tortugarum, behaved in a similar fashion. S. tortugarum do trade eggs, and the phenomenon seems to be widespread in simultaneously hermaphroditic serranids. However, the cues for spawning are much subtler than in H. nigricans and other simultaneously hermaphroditic serranids. This trait appears to be an evolutionary response to the occurrence of heavy sperm competition in the form of streaking.     

PHENOTYPIC DIVERSIFICATION ACROSS AN ENVIRONMENTAL GRADIENT: A ROLE FOR PREDATORS AND RESOURCE AVAILABILITY ON THE EVOLUTION OF LIFE HISTORIES

Changes in age/size‐specific mortality, due to such factors as predation, have potent evolutionary consequences. However, interactions with predators commonly impact prey growth rates and food availability and such indirect effects may also influence evolutionary change. We evaluated life‐history differences in Trinidadian killifish, Rivulus hartii, across a gradient in predation. Rivulus are located in (1) “high predation” sites with large piscivores, (2) “Rivulus/guppy” sites with guppies, and (3) “Rivulus‐only” sites with just Rivulus. Rivulus suffer higher mortality with large predators, and guppies may prey upon small/young Rivulus in Rivulus/guppy environments. In turn, population densities decline while growth rates increase in both localities compared to Rivulus‐only sites. To explore how the direct and indirect effects of predators and guppies influence trait diversification in Rivulus, we examined life‐history phenotypes across five rivers. High predation phenotypes exhibited a smaller size at reproduction, a greater number of eggs that were smaller, and increased reproductive allotment. Such changes are consistent with a direct response to predation. Rivulus from Rivulus/guppy sites were intermediate; they exhibited a smaller size at reproduction, increased fecundity, smaller eggs, and larger reproductive allotment than Rivulus‐only fish. These changes are consistent with models that incorporate the impacts of growth and resources.     

The contribution of aromatic components in Katsuobushi to preference formation and reinforcement effect

Katsuodashi, a dried bonito broth, is very basic and indispensable in Japanese cuisine and contains taste-exhibiting components and unique aroma. We previously reported that its unique aroma contributes to the preference and reinforcement effect associated with dried bonito. This study aims to elucidate the contribution of aromatic components in Katsuobushi to preference formation and reinforcement effect. Volatile components obtained from dried bonito were fractionated and the fractions were subjected to two-bottle choice test. The fractionation test suggested that the component responsible for the preference is not one but comprises multiple components. In the GC–MS analysis/reconstruction test, solution with aromatic flavor narrowed down to 125 compounds had preference, and also had reinforcement effect. Moreover, GC–MS–olfactometry analysis narrowed down the candidate components to 28 out of 125. Mice showed preference for the test solution with aromatic flavor reconstructed with 28 components but did not show reinforcement behavior.     

SENSORY EVALUATION OF VANILLA-DAIRY DESSERTS BY REPERTORY GRID METHOD AND FREE CHOICE PROFILE

“Natillas” are semisolid dairy desserts widely consumed in Spain, with notable differences existing between commercial brands in their composition and sensory and physical properties. The repertory grid method, in conjunction with the free choice profile (FCP), was used to provide data on how consumers perceived eight commercial “natillas” samples. The most frequent terms used were: vanilla flavor, sweetness, consistency and liquid texture as described by 77, 67, 57 and 53% of the consumers, respectively. A three‐dimensional solution was chosen. Dimension 1 accounted for 27.23% of variance and separated the samples largely by yellow‐color intensity and consistency. Dimension 2 (15.88%) was mainly related to the visual attributes of texture, creaminess and different notes of flavor. The third dimension (11.34%) was related to structural‐texture attributes, orange‐yellow color and acid and artificial flavors. The assessors plot showed two clusters. Analysis of each group data revealed differences in their perception of sample variation, mainly regarding texture and color.     

FLAVOR PROPERTIES OF PLAIN SOYMILK*

A trained sensory panel was used to (1) establish terminology for describing flavor attributes of a wide range of “fresh” and processed soymilks (SMs); and (2) test the lexicon and describe the flavor properties of a wide range (n = 32) of plain SM samples. Twenty‐eight attributes were identified in the SMs studied. However, findings indicate that the main differences are a consequence largely of the presence of certain attributes that are either infrequently present or unique to a specific product. Thus, sensory evaluation of SM needs to include a review of samples to ensure that all attributes are documented prior to evaluation, or use a method that allows the addition of attributes during testing.     

Enhancement of cheese flavors with microbial esterases

The characteristic flavor of hard Italian cheeses is associated with the presence of fatty acids, particularly butyric acid, liberated from milk fat during the ripening process. To ensure proper development and control of flavor, animal pregastric esterases or lipases are routinely added to the milk before coagulation of the curd. Such esterases are also used to generate flavor in enzyme modified cheese and other dairy products. Esterases from microbial sources have been investigated as agents to enhance flavor in cheese. We have found that an esterase from Mucor miehei exhibits the type of lipolytic activity needed for this application. Romano and fontina cheeses of excellent quality have been prepared by the use of this esterase. It has also been used successfully in the preparation of enzyme modified cheese, and, in turn, processed American cheese.     

Biotechnological methods to accelerate cheddar cheese ripening

Cheese is one of the dairy products that can result from the enzymatic coagulation of milk. The basic steps of the transformation of milk into cheese are coagulation, draining, and ripening. Ripening is the complex process required for the development of a cheese's flavor, texture and aroma. Proteolysis, lipolysis and glycolysis are the three main biochemical reactions that are responsible for the basic changes during the maturation period. As ripening is a relatively expensive process for the cheese industry, reducing maturation time without destroying the quality of the ripened cheese has economic and technological benefits. Elevated ripening temperatures, addition of enzymes, addition of cheese slurry, attenuated starters, adjunct cultures, genetically engineered starters and recombinant enzymes and microencapsulation of ripening enzymes are traditional and modern methods used to accelerate cheese ripening. In this context, an up to date review of Cheddar cheese ripening is presented.     

Climate mitigation by dairy intensification depends on intensive use of spared grassland

Milk and beef production cause 9% of global greenhouse gas (GHG) emissions. Previous life cycle assessment (LCA) studies have shown that dairy intensification reduces the carbon footprint of milk by increasing animal productivity and feed conversion efficiency. None of these studies simultaneously evaluated indirect GHG effects incurred via teleconnections with expansion of feed crop production and replacement suckler‐beef production. We applied consequential LCA to incorporate these effects into GHG mitigation calculations for intensification scenarios among grazing‐based dairy farms in an industrialized country (UK), in which milk production shifts from average to intensive farm typologies, involving higher milk yields per cow and more maize and concentrate feed in cattle diets. Attributional LCA indicated a reduction of up to 0.10 kg CO₂e kg^?1 milk following intensification, reflecting improved feed conversion efficiency. However, consequential LCA indicated that land use change associated with increased demand for maize and concentrate feed, plus additional suckler‐beef production to replace reduced dairy‐beef output, significantly increased GHG emissions following intensification. International displacement of replacement suckler‐beef production to the “global beef frontier” in Brazil resulted in small GHG savings for the UK GHG inventory, but contributed to a net increase in international GHG emissions equivalent to 0.63 kg CO₂e kg^?1 milk. Use of spared dairy grassland for intensive beef production can lead to net GHG mitigation by replacing extensive beef production, enabling afforestation on larger areas of lower quality grassland, or by avoiding expansion of international (Brazilian) beef production. We recommend that LCA boundaries are expanded when evaluating livestock intensification pathways, to avoid potentially misleading conclusions being drawn from “snapshot” carbon footprints. We conclude that dairy intensification in industrialized countries can lead to significant international carbon leakage, and only achieves GHG mitigation when spared dairy grassland is used to intensify beef production, freeing up larger areas for afforestation.     

Traditional fermented foods

For centuries, diverse plant and animal materials have been fermented by various bacteria, yeasts, and fungi to make excellent foods. The kinds of micro organisms used in traditional fermentation are restricted to a relatively few genera, including Aspergillus, Rhizopus, Mucor, Actinomucor, Monascus, Saccharomyces, Neurospora, Acetobacler, Bacillus, and Lactobacillus. The two principal advantages of food fermentations over other processes are to add flavor and to prevent spoilage. Fermented fish is a common food in the Orient and may have been the first product made by fermentation. Flavor is especially important in vegetable diets based on bland foods such as rice. Shoyu is the best, known oriental food fermentation, and it is very widely used as a flavoring agent. Be sides this fermentation, there are a large number of additional ones not so well known outside the Orient, whose products serve as seasoning or flavoring agents. Miso and natto are prepared from soybeans in Japan. Sufu is a cheese like product made from soybean milk in China. Tempeh and ontjom are Indonesian foods prepared from soybeans and peanuts, respectively. These food fermentations are discussed with emphasis on how they are produced and the flavor formed.