The Anaer­obe Lab

by Roberto

As a stu­dent of mol­e­c­u­lar ge­net­ics in the 1970s, I "grew up" un­der the im­pres­sion that E. coli was the dom­i­nant bac­terium among the mul­ti­tude of mi­cro­bial species res­i­dent in our large in­tes­tine. Its ease of cul­ti­va­tion made it ap­pear so; prob­a­bly go­ing back to the days of Theodor Es­cherich one hun­dred years be­fore. It's no sur­prise then, that to this day, I re­mem­ber the mo­ment I was shock­ingly in­tro­duced to E. coli's ut­terly mi­nus­cule num­bers rel­a­tive to so many other bac­te­r­ial species in the hu­man gut. It was 2004 and I had just heard a talk by Joel Doré at the 10th ISME meet­ing in Can­cún. The over­whelm­ing ma­jor­ity of our gut mi­crobes were anaer­obes. The ad­vent of cul­ture-in­de­pen­dent meth­ods for as­sess­ing mi­cro­bial com­mu­nity com­po­si­tion, al­ready over a decade old and widely used in en­vi­ron­men­tal mi­cro­bi­ol­ogy by then, had just barely been dis­cov­ered by those in­ter­ested hu­man-as­so­ci­ated mi­crobes. 

The ex­plo­sive growth of the field study­ing the hu­man gut mi­cro­biome has been noth­ing short of rev­o­lu­tion­ary. Yet, as won­der­ful as cul­ture-in­de­pen­dent meth­ods are, there's al­ways the need char­ac­ter­ize the in­di­vid­ual strains of (mostly) anaer­o­bic bac­te­ria. For­tu­nately, there are meth­ods to cul­ti­vate those gut anaer­obes. What fol­lows is the story of the place where many of those meth­ods were de­vel­oped, the Anaer­obe Lab at Vir­ginia Tech. More im­por­tantly, it's the story of the two key in­di­vid­u­als that made that lab a re­al­ity. They both are the gi­ants on whose shoul­ders stand all those work­ing on hu­man in­testi­nal mi­cro­bio­mes. Since it is likely that the younger gen­er­a­tions of mi­cro­bi­ol­o­gists do not know their names, it's my hope to help make sure they are re­mem­bered.

One of the two key play­ers was Wal­ter Ed­ward (Ed) Moore. Af­ter train­ing at Madi­son, WI, he was hired at Vir­ginia Tech, circa 1955, and be­gan what would be­come a long and stel­lar ca­reer study­ing anaer­o­bic bac­te­ria. Build­ing on the pi­o­neer­ing work of Robert Hun­gate on cul­ti­vat­ing anaer­obes, Ed im­proved meth­ods for grow­ing the in­testi­nal mi­crobes of farm an­i­mals. At the time, there were no funds to study hu­man-as­so­ci­ated anaer­obes. But the U.S. De­part­ment of Agri­cul­ture and the cat­tle in­dus­try pro­vided plenty of re­search funds to study anaer­obes of the cow's ru­men. Still, Ed was keen on the hu­man gut anaer­obes and, on the side, had a stu­dent cul­ti­vate bac­te­ria from hu­man fe­ces. By the mid-1960s their ef­forts had led them to suc­cess­fully cul­ti­vate what Ed es­ti­mated to be about 80% of the mi­crobes they ob­served in stool sam­ples.

The sec­ond key player was Lil­lian "Peg" Halde­man¹. Peg's path had taken her from train­ing at Duke in Zo­ol­ogy to a po­si­tion at the CDC in At­lanta, start­ing in 1951. While ob­tain­ing on-the-job train­ing and tak­ing night courses at the Uni­ver­sity of Geor­gia, she be­came an ex­pert in the growth and char­ac­ter­i­za­tion of anaer­o­bic bac­te­ria, fo­cus­ing on Clostrid­ium bot­u­linum. She re­mained in her job at CDC for fif­teen years. Along the way, Peg earned a Ph.D. from Mon­tana State in 1964, based on her work on bot­u­linum toxin. 

Two of the few pho­tographs freely avail­able in the web of Peg Halde­man (source) and Ed Moore (source).

Their two tra­jec­to­ries would join and cat­alyze one of the most im­por­tant ven­tures in the his­tory of anaer­o­bic mi­cro­bi­ol­ogy. You might call this an ac­ci­den­tal meet­ing, I much pre­fer to call it a won­der­ful case of serendip­ity, where two par­tic­i­pants of a chance en­counter had the sagac­ity to rec­og­nize the enor­mous pos­si­bil­i­ties of fu­ture col­lab­o­ra­tion. Or, per­haps, I might also call it a match made in heaven. As it turns out, it was a match made in At­lantic City, New Jer­sey.

At­lantic City was the site of the 1965 Gen­eral Meet­ing of the Amer­i­can So­ci­ety for Mi­cro­bi­ol­ogy. There, Peg and Ed met and shared their knowl­edge, the­o­ries and ex­cite­ment around anaer­o­bic res­i­dents of the hu­man gut. It must have been a mag­i­cal mo­ment. Robert Smib­ert, a Vir­ginia Tech col­league was present and re­called: "It was like two way­ward souls get­ting to­gether. We just sat there and lis­tened to them talk. Be­fore we knew it, they came up with what they wanted to do with anaer­obes. So, Peg left the CDC and came up to the vet sci­ence de­part­ment." ²

Two "way­ward souls" now had a com­mon goal. 

In­deed, Peg left the CDC and was ap­pointed Pro­fes­sor of Bac­te­ri­ol­ogy at Vir­gina Tech in 1966. Work­ing with Ed, they jointly lay the foun­da­tion for what was to be­come the Anaer­obe Lab, which, by 1970 had its own build­ing.

The build­ing hous­ing the orig­i­nal Anaer­obe Lab at Vir­ginia Tech. Source

But still, as late as 1970, there were no funds to di­rectly an­a­lyze the anaer­obes of the hu­man gut. Then, in 1971, a fund­ing op­por­tu­nity came from an un­ex­pected source, NASA. The agency wanted to know if as­tro­nauts' mi­cro­biota changed dur­ing space flight. Peg and Ed's pro­posal to NASA was ap­proved and they re­ceived a one-hun­dred-thou­sand-dol­lar grant (a hu­mungous amount at the time). And within a month – be­cause when it rains it pours – they were also funded by the Na­tional Can­cer In­sti­tute as there were hints that ab­nor­mal gut mi­cro­biota could pro­mote colon can­cer. With great lead­er­ship and well-funded, the Anaer­obe Lab was truly up and run­ning. They never stopped to look back. A decade af­ter join­ing the fac­ulty at Vir­ginia Tech, Peg was named Dis­tin­guished Uni­ver­sity Pro­fes­sor.

In the early years of the lab, Peg and Ed, along with their col­leagues and trainees, made nu­mer­ous key con­tri­bu­tions in their field. In 1972, they pub­lished the au­thor­i­ta­tive "Anaer­obe Lab­o­ra­tory Man­ual," de­scrib­ing their cut­ting-edge method­olo­gies for the iso­la­tion and char­ac­ter­i­za­tion of these dif­fi­cult-to-cul­ti­vate bac­te­ria. Among their many re­search pa­pers of that pe­riod, I find two es­pe­cially in­ter­est­ing. First, a study of the fe­cal flora of twenty healthy Japan­ese-Hawai­ians, in which they de­scribed the re­mark­able species di­ver­sity of the hu­man gut mi­cro­biota. Sec­ond, a study on the mi­cro­biota vari­a­tions in the guts of three as­tro­nauts dur­ing a weeks-long on-Earth sim­u­lacrum of the iso­la­tion and diet the as­tro­nauts would en­counter in the Sky­lab. Therein, they de­scribe dra­matic in­creases in the amount of one anaer­obe, en­dear­ingly known as "B. theta," in in­di­vid­u­als un­der emo­tional stress! Turns out, the as­tro­nauts had be­come an­gered and stressed by a se­ries of equip­ment fail­ures and the lousy diet of the Sky­lab. In­ter­est­ingly, Peg and Ed found sim­i­lar in­creases in B. theta in other stressed in­di­vid­u­als (e.g., in sub­jects of an un­re­lated study who were stressed and an­gered while on a long-term mo­not­o­nous diet). Cu­ri­ous about the vari­a­tions of B. theta in re­sponse to stress, they de­ter­mined the amount of B. theta in the fe­ces of grad­u­ate stu­dents on the day of their pre­lim­i­nary qual­i­fy­ing ex­am­i­na­tion and two weeks af­ter. They de­tected no sig­nif­i­cant vari­a­tion. Les­son to be had? Grad­u­ate stu­dents take note: pre­lims are not all that stress­ful 😉.

All in all, Peg and Ed did cut­ting-edge work on anaer­obes for nearly thirty years (1966–1995) at the Anaer­obe Lab. Their work was not lim­ited to the hu­man in­tes­tine. Dur­ing their last ten years of joint work, they moved on to also carry out pi­o­neer­ing re­search in the oral mi­cro­biome.

On a per­sonal note, twenty years af­ter they met in At­lantic City, Peg and Ed mar­ried in 1985³. Maybe it was, af­ter all, a match made in heaven! Ed re­tired from Vir­ginia Tech in 1995 and died in 1996. Peg re­tired in 1996 and died in 2020. Their legacy was to lay the foun­da­tion for much of the work that to this date ad­vances our knowl­edge of the hu­man mi­cro­biome. In that way, they live on!

1 – On Peg's last name as an au­thor. While Peg's last name was Halde­man, she used the al­ter­na­tive spelling Holde­man in all her pa­pers. In­ter­est­ingly, in 1964, she signed her Ph. D. the­sis as Lil­lian Halde­man Holde­man. Why? We'll never find out. Not that it mat­ters.

2 – One key source of in­for­ma­tion I used for this post was Jes­sica Sny­der Sachs' won­der­ful 2007 book "Good Germs, Bad Germs." An out­stand­ing read in its en­tirety. In the book, Jes­sica de­scribes Peg and Ed's story in much greater de­tail than I have done here. Im­por­tantly, the Robert Smib­ert words I cite are taken from Jessica's book and come from a June 19, 2002 in­ter­view con­ducted by John Hess and stored as part of Vir­ginia Tech's Oral His­tory Project. 

3 – Af­ter her mar­riage, Peg au­thored her pa­pers as Lil­lian Moore.