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Reproductive Span of Caenorhabditis elegans Is Extended by Microbacterium sp. Cover

Reproductive Span of Caenorhabditis elegans Is Extended by Microbacterium sp.

Journal of Nematology
Volume 54 (2022): Issue 1 (February 2022)
By: ,  ,  ,  ,  ,  ,   and    
Open Access
|May 2022
Figures & tables

Figures & Tables

Table 1

Average daily individual brood countsa for C. eleganson three environmental bacterial species and the OP50 control.

Reproductive day numberAcinetobacter sp. CFBb9 (N = 4, n = 29)bSerratia sp. CFBb17 (N = 4, n = 39)bMicrobacterium sp. CFBb37 (N = 3, n = 37)bEscherichia coli OP50 (N = 5, n = 33)b
Eggs laid per individualGreater than zero P (one-sample t-test)Eggs laid per individualGreater than zero P (one-sample t-test)Eggs laid per individualGreater than zero P (one-sample t-test)Eggs laid per individualGreater than zero P (one-sample t-test)
Day 141.24<0.00169.30<0.00135.16<0.001151.69<0.001
Day 2162.34<0.001165.74<0.00147.00<0.00195.39<0.001
Day 344.51<0.00139.23<0.00124.08<0.0012.96<0.001
Day 42.07<0.0010.95<0.00113.59<0.0011.12<0.001
Day 5008.86<0.0010
Day 6006.16<0.0010
Day 73.51<0.001
Day 81.49<0.001
Day 91.02<0.001
Day 100.24<0.001
Day 110.51<0.001
Day 120
Day 130
Average brood250.16275.22141.62251.16
size per wormc
RS (days)d44113.8

a First, an average brood number per individual was calculated per pool, then the average of the pools was reported.

b N, number of pools tested; n, total numbers of individuals tested.

c Average brood size per worm in pools is the sum of all the daily brood sizes.

d RS is the number of days with average daily individual brood counts >0 based on one-sample t-test.

RS, reproductive span.

Figure 1

RS of individual nematodes on Microbacterium sp. CFBb37 and the E. coli OP50 control. (A) C. elegans, (B) C. briggsae, (C) C. tropicalis, and (D) Protorhabditis sp. on CFBb37 had longer RS than on OP50 (P < 0.001, log-rank test). Full RS data in Table 2. RS, reproductive span.

Table 2

Total average brood and RS of tested nematode species.

Reproductive day numberOP50CFBb37Different between the strains P (paired t-test)
Eggs laid per day (mean ± SE)Greater than zero P (one-sample t-test)Eggs laid per day (mean ± SE)Greater than zero P (one-sample t-test)
C. elegans
Day 132.68 ± 5.04<0.0013.76 ± 1.74<0.001<0.01
Day 2149.75 ± 4.71<0.00124.64 ± 2.53<0.001<0.001
Day 378.55 ± 6.46<0.00123.49 ± 2.12<0.001<0.001
Day 43.25 ± 0.81<0.00122.05 ± 2.31<0.001<0.001
Day 51.40 ± 0.77>0.0515.10 ± 1.75<0.001<0.001
Day 60.58 ± 0.55>0.057.93 ± 1.14<0.001<0.001
Day 70.05 ± 0.03>0.054.39 ± 0.79<0.001<0.001
Day 80.03 ± 0.03>0.051.88 ± 0.51<0.001<0.01
Day 900.56 ± 0.23<0.05N.A.
Day 100.07 ± 0.04>0.05N.A.
Day 110.03 ± 0.03>0.05N.A.
Day 120
na4041
Average brood size per wormb266.28 ± 6.42137.63 ± 5.74<0.001
RS (day)c4.08 ± 0.167.29 ± 0.25<0.001 (log-rank test)
C. briggsae
Day 111.59 ± 1.33<0.0012.11 ± 0.34<0.001<0.001
Day 218.21 ± 1.89<0.0013.80 ± 0.57<0.001<0.001
Day 322.56 ± 3.46<0.0013.17 ± 0.56<0.001<0.001
Day 412.00 ± 2.52<0.0012.03 ± 0.50<0.001<0.001
Day 50.88 ± 0.28<0.012.30 ± 0.50<0.0010.0109
Day 60.16 ± 0.10>0.051.69 ± 0.44<0.001<0.01
Day 700.50 ± 0.17<0.001N.A.
Day 80.22 ± 0.07<0.01N.A.
Day 90.08 ± 0.08>0.05N.A.
Day 100.08 ± 0.08>0.05N.A.
Day 110
na3236
Average brood size per wormb65.40 ± 7.0916.00 ± 1.88<0.001
RS (day)c3.97 ± 0.255.61 ± 0.39<0.001 (log-rank test)
C. tropicalis
Day 14.37 ± 0.78<0.0014.75 ± 0.65<0.0011.0
Day 24.81 ± 1.37<0.0014.79 ± 0.92<0.0010.8652
Day 31.22 ± 0.39<0.017.20 ± 1.71<0.001<0.01
Day 40.37 ± 0.24>0.053.88 ± 1.05<0.01<0.01
Day 503.29 ± 0.92<0.01N.A.
Day 60.95 ± 0.29<0.01N.A.
Day 70.33 ± 0.14<0.05N.A.
Day 80.04 ± 0.04>0.05N.A.
Day 90N.A.
na2724
Average brood size per wormb10.78 ± 1.9025.25 ± 3.59<0.01
RS (day)c2.37 ± 0.204.96 ± 0.36<0.001 (log-rank test)
Protorhabditis sp.
Day 145.00 ± 5.02<0.0012.70 ± 0.30<0.001<0.001
Day 295.15 ± 7.18<0.0014.21 ± 0.49<0.001<0.001
Day 318.06 ± 4.95<0.0013.23 ± 0.52<0.001<0.01
Day 42.00 ± 0.69<0.013.12 ± 0.56<0.001>0.05
Day 50.88 ± 0.70>0.052.38 ± 0.43<0.001N.A.
Day 60.24 ± 0.18>0.051.36 ± 0.28<0.001N.A.
Day 70.06 ± 0.06>0.051.09 ± 0.24<0.001N.A.
Day 801.00 ± 0.24<0.001N.A.
Day 91.17 ± 0.33<0.01N.A.
Day 100.53 ± 0.18<0.01N.A.
Day 110.53 ± 0.19<0.01N.A.
Day 120.42 ± 0.14<0.01N.A.
Day 130.14 ± 0.06<0.05N.A.
Day 140.10 ± 0.07>0.05N.A.
Day 150.06 ± 0.04>0.05N.A.
0
na3447
Average brood size per wormb161.38 ± 10.0522.08 ± 2.26<0.01
RS (day)c3.59 ± 0.206.68 ± 0.61<0.001 (log-rank test)

a n, the total number of tested individuals in each test.

b Average brood size per worm was the total brood of all hermaphrodites (n) divided by n.

c Average RS. N.A., not applicable.

RS, reproductive span; SE, standard error.

Figure 2

Lifespan assays of C. elegans on Microbacterium sp. CFBb37 and the OP50 control. C. elegans grown on CFBb37 survived longer than OP50 (P < 0.001, log-rank test). Detailed lifespan data are in Tables S4 and S5.

Figure 3

Summary of RS extension in the Caenorhabditis species (orange) and the outgroup Protorhabditis sp. (light blue). Red lines indicate inferred ancestral evolutionary lineages with extended RS. We suggest that other species within the genus Caenorhabditis are likely to have extended RS but this is untested. For simplification, some species are not listed and are grouped together in triangles, with the numbers of known species indicated. Phylogeny is adapted from Kiontke and Fitch (2005) and Felix et al. (2014) with the position of Protorhabditis sp. CFB231 placed next to P. sp. 1 species based on the 18S ribosomal RNA gene sequence (Zhang et al., 2000; Morgulis et al., 2008). Protorhabditis sp. CFB231 is self-fertile but the sexual system, hermaphroditic or parthenogenetic, is unknown (indicated by a question mark). RS, reproductive span.

Figure S1

Size comparison of the four tested nematode species grown on Microbacterium sp. CFBb37 and Escherichia coli OP50. Images are representative samples.

Figure S2

Protorhabditis sp. CFB231 make sperm. (A) DIC image of spermathecal region (outlined). One oocyte is apparently transiting the spermatheca. (B) DAPI staining for nuclei. (C) Merged images of DIC and DAPI. DIC, differential interference contrast.

Table S1

Wild-type isolates of nematodes in Cat Tien National Park.

Species nameStrain numberGenBank accession No.Best BLAST hitaQuery coverageIdentity (%)Bit score (max/total)E value
Caenorhabditis briggsaeCFB233MZ457559Caenorhabditis briggsae AF1698%100%1,544/1,5440.0
Protorhabditis sp.CFB231MZ474671Protorhabditis sp. 1 GVDU-201990%95.351,229/1,2290.0

a Query date March 25, 2022.

Table S2

Bacterial strains isolated from rotting vegetation samples collected in Cat Tien National Park and their effects on nematodes.

StrainaSpeciesIsolation siteGenBank accession No.Phenotype
CFBb3Pseudomonas sp.11°23′54.4″N
107°20′21.4″E		OM456346Late laying egg
CFBb9bAcinetobacter sp.11°24′20.9″N
107°24′22.3″E		MZ467312Normal
CFBb11Acinetobacter sp.11°24′20.9″N
107°24′22.3″E		OM456348Small body
CFBb17Serratia sp.11°24′21.4″N
107°24′21.4″E		MZ457333Small body
CFBb35Serratia sp.11°27′5″N
107°27′25.6″E		OM629173Normal
CFBb37Microbacterium sp11°27′5″N
107°27′25.6″E		MZ453404RS; small body
CFBb39Sphingobacterium sp.11°27′17.9″N
107°22′5.6″E		OM892000Normal
CFBb40Bacillus sp.11°27′17.9″N
107°22′5.6″E		OM456347Normal
CFBb42Serratia sp.11°26′47.8″N
107°26′19.8″E		OM456350Small body
CFBb53Bacillus cereus11°26′12.1″N
107°25′25.7″E		OM883917Small body
CFBb54Enterobacter sp.11°26′12.1″N
107°25′25.7″E		OM456349Small body
CFBb57Serratia sp.11°23′26″N
107°21′1.8″E		OM892003Small body

Gray highlights bacteria isolated from the same sites as the two new nematode strains isolated from this study (C. briggsae CFB233 [light gray] and Protorhabditis sp. CFB231 [dark gray]).

a Strains were isolated in this study unless indicated.

b From Le et al., 2021.

RS, reproductive span.

Table S3

Bacterial isolates with description of colonial morphology, and geography of sample sites.

Cat Tien National ParkCuc Phuong National Park
StrainMorphology of colonyaNorthEastStrainMorphology of colonyaNorthEast
CFBb1Small11°23′57.5″107°20′21.4″CFBb65Big20°17′40.4″105°39′46.6″
CFBb2Big11°23′54.4″107°20′21.4″CFBb66Big––
CFBb3Small––CFBb67Small––
CFBb4Irregular11°24′22.8″107°24′22″CFBb68Small––
CFBb5Irregular––CFBb69Big20°17′40″105°39′57.5″
CFBb6Big––CFBb70Small––
CFBb7Big––CFBb71Small20°17′41.3″105°39′59.6″
CFBb8Small––CFBb72Small––
CFBb9Big11°24′20.9″107°24′22.3″CFBb73Big––
CFBb10Irregular11°24′21.4″107°24′25.6″CFBb74Big––
CFBb11Big11°24′20.9″107°24′22.3″CFBb75Small––
CFBb12Small11°24′20.9″107°24′22.1″CFBb76Irregular20°20′56.7″105°35′48.1″
CFBb13Big11°24′19.6″107°24′22.1″CFBb77Big––
CFBb14Small––CFBb78Small20°15′51.2″105°41′57.6″
CFBb15Big11°24′19″107°24′22.2″CFBb79Big20°20′56.6″105°35′47.4″
CFBb16Big11°24′18.6″107°24′22.3″CFBb80Small––
CFBb17Irregular11°24′21.4″107°24′21.4″CFBb81Irregular––
CFBb18Big––CFBb82Big20°15′7.6″105°42′45″
CFBb19Big11°28′46.4″107°22′52.9″CFBb83Small––
CFBb20Small––CFBb84Big20°14′53.5″105°42′34.3″
CFBb21Medium11°23′18.7″107°21′3.4″CFBb85Small––
CFBb22Medium––CFBb86Small––
CFBb23Big11°29′7.2″107°22′58″CFBb87Medium20°14′59.8″105°42′28.1″
CFBb24Small––CFBb88Medium––
CFBb25Medium11°27′56.1″107°22′43.5″CFBb89Big––
CFBb26Big, yellow11°27′0.9″107°21′27.1″CFBb90Small––
CFBb27Small––CFBb91Big20°14′56.7″105°42′21.1″
CFBb28Big11°27′1.5″107°21′27.5″CFBb92Big––
CFBb29Small––CFBb93Big20°14′39.1″105°42′25.3″
CFBb30Big, red11°26′56.6″107°21′35.9″CFBb94Small––
CFBb31Big––CFBb95Big20°20′56.7″105°35′46.5″
CFBb32Big11°23′18.8″107°21′3.6″CFBb96Small––
CFBb33Small––CFBb97Medium20°20′58.4″105°35′36.5″
CFBb34Big11°27′31.9″107°20′43.1″CFBb98Medium––
CFBb35Big11°27′5″107°27′25.6″CFBb99Big––
CFBb36Irregular11°27′3″107°21′51.7″CFBb100Big––
CFBb37Small11°27′5″107°27′25.6″CFBb101Small––
CFBb38Small––CFBb102Big20°21′0.5″105°35′36.3″
CFBb39Big11°27′17.9″107°22′5.6″CFBb103Big––
CFBb40Big11°27′17.9″107°22′5.6″CFBb104Small––
CFBb41Small11°27′17.9″107°22′5.6″CFBb105Small––
CFBb42Big11°26′47.8″107°26′19.8″CFBb106Big––
CFBb43Small11°26′47.8″107°26″19.8″CFBb107Small––
CFBb44Big11°27′50.4″107°27′43.5″CFBb108Big20°20′54.1″105°35′48.5″
CFBb45Irregular11°27′51.8″107°27′42.1″CFBb109Small––
CFBb46Big––
CFBb47Small––
CFBb48Big11°23′19.3″107°21′3.7″
CFBb49Small––
CFBb50Big, pink11°26′14.5″107°25′24.2″
CFBb51Small––
CFBb52Big11°26′12.1″107°25′25.7″
CFBb53Small11°26′12.1″107°25′25.7″
CFBb54Small11°26′12.1″107°25′25.7″
CFBb55Big11°25′23″107°25′41.4″
CFBb56Small––
CFBb57Medium11°23′26″107°21′1.8″
CFBb58Big11°24′49″107°25′27.7″
CFBb59Big––
CFBb60Small––
CFBb61Small––
CFBb62Big––
CFBb63Medium11°24′48.2″107°25′25.9″
CFBb64Small11°23′39.7″107°21′47.9″

Dashes indicate the same site above.

a Colonies had circular shapes and white color, otherwise noted as “irregular” and specific color.

Table S4

Counts of surviving and lifespan of C. elegans on Microbacterium sp. CFBb37 and the OP50 control.

Day of lifespanNumber of surviving worms
OP50CFBb37
17360
27360
37360
47360
57360
67360
77360
87360
97259
107259
116959
126659
136358
144654
153953
163652
172552
181849
191648
201448
211347
221247
231145
24743
25643
26642
27436
28330
29325
30124
31116
32011
336
341
350
Censored70
Lifespan (days) (mean ± 1SE)17.26 ± 0.5726.77 ± 0.86
P (log-rank test)<0.001

[i] SE, standard error.

Table S5

Counts of daily uterus eggs of close nematode species.

NematodeReproductive day numberOP50CFBb37Different between the strains P (paired t-test)
Number of tested hermaphroditesNumber of eggs in uterus/individual (mean ± SE)Greater than zero P (one-sample t-test)Number of tested hermaphroditesNumber of eggs in uterus/individual (mean ± SE)Greater than zero P (one-sample t-test)
C. elegansDay 1323.94 ± 0.68<0.001351.57 ± 0.34<0.0010.0051
Day 23225.25 ± 1.12<0.001313.13 ± 0.32<0.001<0.001
Day 3295.38 ± 0.98<0.001424.02 ± 0.33<0.0010.06615
Day 4301.13 ± 0.34<0.01323.69 ± 0.33<0.001<0.001
Day 5370.03 ± 0.03>0.05343.5 ± 0.46<0.001<0.001
Day 6373.22 ± 0.51<0.001
Day 7342.29 ± 0.36<0.001
Day 8350.43 ± 0.16<0.01
C. briggsaeDay 1393.30 ± 0.73<0.001390.51 ± 0.09<0.001<0.001
Day 2
Day 3344.09 ± 0.39<0.001350.6 ± 0.15<0.001<0.001
Day 4392.38 ± 0.24<0.001320.81 ± 0.14<0.001<0.001
Day 5350.2 ± 0.07<0.01351.46 ± 0.15<0.001<0.001
Day 6340.09 ± 0.05>0.05311.45 ± 0.19<0.001<0.001
Day 7320.63 ± 0.15<0.001
C. tropicalisDay 1332.09 ± 0.22<0.001320.28 ± 0.09<0.001<0.001
Day 2323.47 ± 0.23<0.001372.76 ± 0.32<0.0010.2149
Day 3381.87 ± 0.23<0.001363.19 ± 0.4<0.0010.0044
Day 4280.64 ± 0.23<0.05382.6 ± 0.24<0.001<0.001
Day 5360.19 ± 0.13>0.05342.94 ± 0.36<0.001<0.001
Day 6361.53 ± 0.2<0.001
Day 7271.63 ± 0.22<0.001
Day 8170.82 ± 0.16<0.01
P. sp.Day 1343.23 ± 0.55<0.001323.28 ± 0.26<0.0010.595
Day 2363.67 ± 0.63<0.001381.61 ± 0.18<0.001<0.001
Day 3391.38 ± 0.24<0.001361.58 ± 0.12<0.0010.629
Day 4360.19 ± 0.03<0.05361.56 ± 0.12<0.001<0.001
Day 5340.15 ± 0.03>0.05401.80 ± 0.16<0.001<0.001
Day 6351.06 ± 0.13<0.001<0.001
Day 7530.81 ± 0.10<0.001<0.001

[i] SE, standard error.

DOI: https://doi.org/10.2478/jofnem-2022-0010 | Journal eISSN: 2640-396X | Journal ISSN: 0022-300X
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Language: English
Submitted on: Jun 29, 2021
Published on: May 10, 2022
Published by: Society of Nematologists, Inc.
In partnership with: Paradigm Publishing Services
Publication frequency: 1 issue per year
Keywords:
bacteria,
brood size,
Caenorhabditis nematodes,
lifespan,
Microbacterium sp.,
physiology,
Protorhabditis sp.,
reproductive span
Related subjects:
Life sciences,
Life sciences, other

© 2022 Tho Son Le, Thi Hong Gam Nguyen, Bich Hong Ha, Bui Thi Mai Huong, Thi Thu Hang Nguyen, Kim Dung Vu, Tu Cuong Ho, John Wang, published by Society of Nematologists, Inc.
This work is licensed under the Creative Commons Attribution 4.0 License.

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