Penicillin

The discovery of antibiotics is one of the most significant medical accomplishments of the last century. Their impact has been profound. Along with improvements in sanitation and vaccinations, the use of antibiotics remains at the forefront of our largely successful efforts to control infectious diseases. As with PCR, we owe the availability of antibiotics to evolution. They are not invented, they are produced naturally by a variety of different organisms.

The modern era of control of bacterial infections began in 1927 with the discovery by Alexander Fleming that extracts from the mold Penicillium notatum lysed bacterial cells. He was not able to isolate the active compound but gave it the name Penicillin after the mold’s genus. The active compound was eventually isolated and developed into a therapy in the early 1940’s by a groups led by Chan and Florey. For their efforts, they shared the 1945 nobel prize for physiology and medicine with Fleming. The receipt of the Nobel Prize just a few short years after the introduction of penicillin as a drug speaks to the impact this medication had.

Many new antibiotics have been introduced since the 1940's but Penicillin and penicillin like antibiotics remain in wide use, comprising almost 20% of all antibiotics manufactured.

Image source

Flavobacterium columnare

I study the bacteria Flavobacterium columnare. It is a fresh water bacteria found all over the world. It is an opportunistic fish pathogen and is the cause of Columnaris Disease (CD). It infects many commercially important fish species. Outbreaks in aquaculture facilities are common. Costs associated with CD are high and represent a major barrier to the development of commercially viable aquaculture enterprises.

Outbreaks are brought on by stress and can occur suddenly. Mortality rates can reach 100%. The most virulent strains F. columnare kill susceptible fish within 24 to 48 hours, leaving little time for treatment.

CD is common in tropical aquaria where it is called by various names including, fin rot, tail rot and cotton mouth. These are frequently discussed on websites for aquaria hobbyist. The bacteria responsible is often identified as Flexibacter columnaris, an obsolete genus and species name replaced in the mid 1990. The characteristic fuzzy white growth the bacteria cause, often results in hobbyists mistaking it for a fungal infection.

CD also effects wild fish stocks. Outbreaks usually occur in the spring as the water warms. These outbreaks can be quite dramatic with hundreds or even thousands of moribund fish washing ashore covered with lesions. In less populous areas, these outbreaks go unnoticed. When they occur in lakes in populated areas, the fish die-offs make the news.

No one likes to see (or smell) tons of dead fish washing up on their shore.

Bacteria and the cost of oil

Oil is considered sour if it contains reduced sulfur compounds (sulfides) at concentrations of 1% or greater. High concentrations of sulfides in oil are problematic for a variety of reasons including:

• Hydrogen sulfide is extremely corrosive and can cause damage to the pipes used to transport oil.
  
• When complexed with other metals such as iron, the sulfides can form precipitates that restrict the flow of oil in the pipes.
  
• Sulfides are toxic and cause environmental and health problems in areas where sour oil is produced, processed or burned.

Removal of sulfides is costly and so refining costs for sour oil is significantly greater than for sweet oil.

Souring of oil is exacerbated by the common practice of pumping water into older oil fields to increase the pressure in the fields as a way to increase oil recovery. Depending on the source of the water used, this practice can introduce large quantities of sulfate (SO₄) into the oil/water mixture. Any oxygen present in the water when it is first pumped underground is rapidly consumed by microbial activity. Once the oxygen is gone, anaerobic microbes can contiune to extract energy from the organic mater present by using compounds other than oxygen as terminal electron acceptors. Sulfate reducing bacteria or SRBs are anaerobes that are able to use sulfate as an electron acceptor. The process results in the production of oxidized carbon compounds and reduced sulfur (sulfides). Biocides are often added to the water to inhibit microbial activity but this process is not efficient requiring enormous amounts of toxic compounds to be added to the water to have a lasting impact.

Thus, microbial activity in oil fields contributes the the cost of oil production. Microbial sulfide production is not limited to oil reservoirs. SRBs are also responsible for the 'rotten egg' smell associated with other anaerobic environments such as swamps and septic systems.

Stability - Diversity relationships

I mentioned in this post, my concerns about speculations by Xu et al on the role of the human host in the maintenance of a diverse gut microbial community. The proposed benefit to us is that the high diversity encouraged stability and assured that our guts continued to provide the desired services, but the mechanism by which we control diversity was not clear.

An article by Ives and Carpenter in a recent issue of the journal Science makes it clear that Xu et al. are in good company. Ives and Carpenter state that we lack of a good understanding of the relationship between diversity and stability in part because term stability is actually used in several related (but distinct) ways in the ecology literature.

Understanding the dynamics of complex systems such as the human gut is challenging. Here is the background knowledge Ives and Carpenter suggest is necessary for beginning to develop an understanding of the diversity/stability relationship:

Before designing an empirical study, it is necessary to know enough about the dynamics of an ecosystem and the environmental perturbations that impinge upon it to select appropriate definitions of stability; there will often be several appropriate definitions. These concepts also identify key features—we will refer to them as mechanisms—that together dictate stability. These mechanisms involve the strength of interactions among species, the mode in which species interact (whether they are competitors, predators, mutualists, etc.) that gives the food-web topology, and the ways in which species experience different types of environmental perturbations. Because both species interactions and environmental perturbations can drive fluctuations in species densities, these must be sorted out and quantified to understand their mechanistic roles in diversity-stability relationships.

And, here is a excerpt from the recommendations they make at the end of the paper:

The relationship between diversity and stability has interested ecologists since the inception of the discipline (35), and the absence of a resolution reflects the complexity of the problem. Much of the complexity derives from the multiplicity of diversity-stability relationships, depending on the definitions of diversity and stability and on the context in which an ecosystem is perturbed. We cannot expect a general conclusion about the diversity-stability relationship, and simply increasing the number of studies on different ecosystems will not generate one.

Rather than search for generalities in patterns of diversity-stability relationships, we recommend investigating mechanisms. A given diversity-stability relationship may be driven by multiple mechanisms, and the same mechanisms may evoke different diversity-stability relationships depending on the definitions of diversity and stability. We need more studies revealing exactly what these mechanisms are. This requires models joined to empirical studies that can reproduce, in a statistically robust way, not only a diversity-stability relationship but also the dynamics exhibited by a system.

The human gut community does exhibit characteristics of a stable system such as the ability to resist perturbations. So, what are the mechanisms that maintain the diversity, what is the diversity stability relationship and how do we go about studying it.

More on human guts

Yet another interesting open access gut microbe paper in PLoS Biology came out in June. This one describes patterns in the colonization of the intenstines of human infants. As mentioned in a previous post, we are born with a sterile intestinal tract and depend upon the ingestion of compatible microbes for the establishment of our gut community. This study used 16S rhibosomal DNA sequences to document changes in the structure and diversity of infant guts over the first year of life. As with the previous paper, Liza Gross wrote a nice summary article.

Some key points:

• 14 babies were followed (including one set of twins) for one year. Early the communities were quite different but by the end of the first year they had acquired a composition similar to that of the adult human.
• At one week of age, two babies delivered by cesarian had fewer total gut bacteria indicating that during natural child birth, the colonization begins during the birthing process.
• While broadly similar to each other and to the adult community, each infant had a distinct profile that persisted over time.
  

This paragraph from the end of the Gross summary provides a good overview of the most interesting findings:

The idiosyncratic nature of the early stages of colonization suggests that a baby’s initial bacterial profile largely results from incidental microbial encounters. The fact that some of the early stool samples matched their mother’s breast milk or vaginal sample supports this interpretation. Shared environment may also explain the coincidental appearance of microbes in the twins. The researchers explain the tendency of these communities to eventually converge by hypothesizing that the human–microbe symbiosis has likely evolved under strong selection and that certain well-adapted microbes repeatedly “win” the battle over the opportunistic early colonizers.

Selections from the final paragraph describes some of the future directions the work will take:

By comparing the surprising range of microbial profiles found in these healthy babies to the microbiota of infants born prematurely or with health problems, future studies can explore how diet, delivery method, or other factors might spell the difference between health and disease.

and that the approach used in the study will allow us to explore questions about

the environmental and genetic factors that shape and personalize the amazing “alien” ecosystem that lives within us.

Bacteriorhodopsin phototrophy

Classification of the metabolic capabilities of microbes can be challenging. With few exceptions, macroorganisms are either photosynthesizing primary producers (photo-autotrophs) or consumers (organotrophs or more commonly, heterotrophs).

For microbes, the story is more complicated. In addition to phototrophy, microbes can be chemo- or litho-trophs meaning they are able to derive energy from the oxidation of inorganic compounds such as reduced sulfur. If they can use light or chemical energy to fix carbon, then they are considered autotrophs. If the energy they acquire can be used to synthesize ATP but not to fix carbon, they are dependent on external sources of organic carbon making them mixotrophs.

An example of a phototrophic mixotroph is pictured above. These are salt loving haloarchaea in salt production ponds near (in?) San Francisco. The red color is due to the transmembrane protein bacteriorhodopsin. Using this protein some haloarchaea can harness sunlight to pump protons across their cell membrane. This establishes a proton gradient across the membrane. This gradient can be used to generate ATP.

There is a large number of scientific papers on bacteriorhodopsin because of its relative simplicity, it has become a model system for the study of membrane associated ion pumps.

Image from here http://www.sfgate.com/cgi-bin/article.cgi?file=/chronicle/archive/2001/03/13/MN152047.DTL

These salt ponds are near San Francisco. If you want another view of the bay area ponds follow this link, select the satellite map and zoom way in. I tried this for a few of the other places I know these salt production ponds exist but the satellite images did not provide good enough resolution. An example is Bon Aire in the Netherlands Antillies

Cilliate kleptoplasty

Chloroplasts, like mitochondria, have their own chromosomal DNA. This is, of course, evidence for the endosymbiotic theory of the origin of chloroplasts. It is also useful because it allows researchers to use the DNA to identify the source of the chloroplasts present in kleptoplastic organisms. This is a fairly standard method and is the way that Gast et al 2007 determined the source of the chloroplasts in the Antartic dinoflagellates discussed in a previous post.

In a paper (pdf) a few years ago in Limnology and Oceanography, McManus et al. used the chloroplasts present in the kleptoplastic tide-pool ciliates, Strombidium oculatum and Strombidium styliferseen to help reveal an interesting life history. The chloroplasts were from the large multicellular macroalgae Enteromorpha clathrata which raised the question of how these unicellular cilliates were able to acquire macroalgal chloroplasts.

McManus et al. found that the cilliates don't appear to be grazing directly on the large strands of the mature algae but on the small motile reproductive cells, called zoospores, mature algal strands release.

In addition to chloroplasts, the zoospores contain a pigmented eyespot. As the photo above (from the paper's figure 1) demonstrates, the kleptoplastic cilliates contain a pigmented eyespot similar to the ones possessed by the zoospores. This suggests that the Strombidium cilliates also owe their phototaxic abilities to the alga cells they ingest.

Some other interesting points about these cilliates:

• They are tidal organisms and live by the rhythm of the tides, becoming active during low tide when tidal pools are calm, and then attach to surfaces and encyst during high tide, presumably to prevent them from being washed out to sea and away from their food.
  
• They appear to be obligate mixotrophs, unable to grow in the dark or in the absence of algal food.

Complete reference:
McManus, G. B., H. Zhang, and S. Lin. 2004. Marine planktonic ciliates that prey on macroalgae and enslave their chloroplasts. Limnol. Oceanogr. 49:308-313.

Cow burps and global warming

Methane is a greenhouse gas and methane released into the atmosphere contributes to global climate change. Compared to CO₂, relatively little methane is released but it is ~ 20 times more effective at absorbing energy than CO₂ so its contribution is significant. There are both human influenced and natural sources of methane. These including rice patties, swamp gasses, termites and ruminants such as cows.

My friend (and a reader of this blog) Marek Kirs send me this link to a story on NPR about attempts to reduce the amount of methane produced by cows. The idea is to adjust the types of food they eat or even manipulate their gut microbial community to eliminate the methanogenic organisms responsible for the gas production.

One of the points the researchers make is that the production of methane is the result of incomplete oxidation of food eaten by the cows. So, the elimination of methanogenesis could result in increases production of meat and milk from these animals. Cattle food conversion ration are around 12%, and any increases would likely be small but the idea is to have enough of an increase to offset any increased cost to the producers.

Gut bacteria metabolic diversity

Another paper exploring microbial diversity of the human gut has been released in PLoS Biology. It is open source so go read it for yourself! There is also a summary article on the data intended for a lay audience. The data presented in the paper is very interesting. They sequenced the genomes of two gut bacteria, Bacteroides vulgatus and Bacteroides distasonis, and contrasted their metabolic potential (the genes present in the genome) with the well studied species Bacteroides thetaiotaomicron. This comparison does provide some evidence that niche specialization plays a role in maintaining the diversity of the gut microbiota.

The importance of studying the gut microbiota is summed up nicely in the beginning of the author summary:

"Our microbial partners provide us with certain features that we have not had to evolve on our own. In this sense, we should consider ourselves to be a supraorganism whose genetic landscape includes both our own genome as well as the genomes of our resident microbes, and whose physiologic features are a synthesis of human and microbial metabolic traits."

I would have worded the first sentence differently as saying 'have not had to" sounds a bit too teleological to me.

I am also troubled by their use (later in the introduction) of the term "top-down selection" in reference to host driven selective forces that they argue are responsible for maintaining a high degree of functional redundancy in the gut community. I am not aware that the term top-down selection, as used by the broader ecology community, is considered a force for the maintenance of ecosystem stability.

The end of the paper lists a set open questions that it would be very nice to have answers to:

"Do we share an identifiable core “microbiome”? If there is such a core, how does the shell of diversity that surrounds the core influence our individual physiologic properties? How is the human microbiome evolving (within and between individuals) over varying time scales as a function of our changing diets, lifestyle, and biosphere? Finally, how should we define members of the microbiome when microbes possess pan-genomes (all genes present in any of the strains of a species) with varying degrees of “openness” to acquisition of genes from other microbes?"