BLANCO, JOSE ENRICO PAOLO L.

OCTOBER 22, 2019

VENTURING THE DIVERSITY, THE PARASITIC AND MUTUALISTIC

EFFECTS, AND THE POTENTIAL APPLICATION OF WOLBACHIA

Wolbachia are classified as members of order Rickettsiales which are

characterized as an intracellular bacteria. It is commonly found in different species of
nematodes and arthropods. Wolbachia genus is divided accordingly into eight
supergroups with monophyletic relationship among each other. Wolbachia is reportedly
transmittable through lateral and vertical gene transfer. In that way, Wolbachia bacteria
can be used in suppressing disease infection, controlling pest population, and reducing
virus transmission (World Mosquito Program, n.d.).
It was mentioned that the genus Wolbachia is divided into eight supergroups with
monophyletic relationship among each other (Casiraghi, et. al., 2005). Based from
Werren, Baldo & Clark, it is recorded that Wolbachia genetic diversity ranges from 6% to
9% in terms of nucleotide divergence in housekeeping genes, and could rise to 30% in
prophage ORF7, and could exceed >43% at wsp (2008). The uncertainty concerning the
naming of Wolbachia has for quite some time been a problem for organizing and
understanding strain diversity. In fact, the discovery of extensive recombination has been
a problem for traditional phylogeny methods. Extensive recombination among strains
affects the genomic characteristic including surface protein, housekeeping, and prophage
genes together with intergenic regions (Baldo, Bordenstein, Wernegreen, & Werren,
2005). It is the primary reason for the acceleration of genetic and functional diversity.
Recombination among Wolbachia is applicable with high genetic diversity in the genus
and the genus cohesiveness of Wolbachia strains with respect to a distant strain.

Wolbachia has the ability to live within and to operate the mechanism on cell and
reproductive process in invertebrates. With that, it reflects their long evolutionary history
as intracellular bacteria. On the study of Baldo, Clark, & Werren, it has been found that
only one other bacterial group, the genus Candidatus Cardinium on the class of
Bacteriodetes, to have an unusual,similar, and diverse set of reproductive manipulation
as to Wolbachia(2008).

Studies had shown that Wolbachia infection causes phenotypical effects including
Cytoplasmic incompatibility, Parthenogenesis induction, feminization, male killing, and
multi-potent Wolbachea.

First, cytoplasmic incompatibility is cause by Wolbachia infection. It is the most

frequently found Wolbachia-induced phenotypical effect and is described by O’Neill,
Giordano, Colbert, Karr, & Robertson as an occurrence in which certain crosses between
symbiont-infected individuals lead to embryonic death or sex ratio distortion (1992). It was
seen commonly in each host taxon there were defects especially in the embryonic mitosis
inferable from the disturbance of cell cycle which result to asynchronous development of
the pronuclei in male and female causing to the incompatible cross (Lassy & Karr, 1996).
Inconsistent crosses bring out haploid development, which has been seen in mosquitoes,
wasps, and flies. In diploid living beings, the outcome will be embryonic lethality, while on
haplodiploids, haploidy will result to normal male development (Reed & Werren, 1995).
Despite that, the molecular mechanism that underlie CI remain unclear despite numerous
studies were conducted on the effect and various proposed mechanisms (Poinsot,
Charlat, & Mercot, 2003).
Second, on the same hand, another cause of Wolbachia infection is
Parthenogenesis induction. Wolbachia-induced female parthenogenesis induction is less
common than cytoplasmic incompatibility and only observed in selected species that
undergoes arrhenotokous development which is define as an observable phenomenon
where unfertilized egg turns into a male organism. Mites, hymenopterans, and thrips are
some common species
that undergo arrhenotokous development (Weeks & Breeuwer, 2001). But in cases
when female was infected, instead of producing a male offspring from an unfertilized egg,
female offspring is produced. Also, infected females can go down the bacteria to their
offspring not at all like for the situation for infected male where it isn't transmitted. In the
study of Stouthamer & Kazmer (1994), it was mentioned that Wolbachia-induced
parthenogenesis is the result of cell cycle disruption during the early embryonic
improvement which is similar to cytoplasmic incompatibility and primarily behind why
unfertilized eggs experience diploid development.
Third, another cause of Wolbachia infection is feminization. Wolbachia-induced
feminization was first seen in species of isopods and was seen lately in species of insects.
Feminization occurs in different mechanism depending on the organism. It was observed
in some isopod species specifically from Oniscidea, wolbachia have been appeared to
multiply inside the androgenic organ, causing to androgenic gland hypertrophy and
inhibited function. Therefore, genetic males turn upon from females because of
feminization event (Vandekerckhove, et al., 2003). Given all information, the mechanism
of the feminization does not have enough evidence to state as it really occurs. In the case
of Eurema hecabe, Wolbachia must be readily existing and functioning throughout the
development for successful feminization. The absence of Wolbachia in the development
will result in intersexual development (Narita, Kageyama, Nomura & Fukatsu,2007).

Fourth, same cause of Wolbachia infection is male killing. This mainly occurring in
arthropod order Coleoptera, Diptera, Lepidoptera, and Pseudoscorpiones. Wolbachia
killing of males happens during embryogenesis; hence, it will result to more food materials
for female progeny. According to Kageyama, Nishimura, Hoshizaki, & Isikawa the reason
for male killing in lepidopteran host namely Ostrinia scapulalis was brought from
feminization induced by Wolbachia. Genetic females die during larval development when
there is an absence of Wolbachia, while in the presence of Wolbachia, feminization
happen to genetic males and eventually die during the stage of larval development.
Hence, Wolbachia-induced male killing happens through lethal feminization (Kageyama
& Traut, 2004).
 Fifth, there are selected strains that can induce multiple phenotype. One evidence
is a Wolbachia strain that infects the lepidopteran host Cadra cautella normally causes
CI, but when cases that the strain transferred into different host, the same strain cause
male killing but not CI (Sasaki & Ishikawa, 2000). Wolbachia has a lot more effect to
discover. Its infection is not limited only for negative effects. It also has its positive effects.

Wolbachia could possibly have implication and application specifically in

combatting human disease and controlling pest (Bourtzis, 2008; Pfarr & Houerauf, 2006).
It contributes to resolving the filarial pathogenicity (Saint André, 2002). Studies have
shown interest on Anti- Wolbachea therapies due to its promising effect in eliminating
filarial nematode infections and in lessening the effect of infections. Antibiotic intervention
in reducing nematode load is not enough (Richards Jr., 2007). Anti-Wolbachia therapy
treatment together with traditional vermicides is an effective tool in resolving the problem
(Debrah, et. al., 2007; Debrah, et. al., 2006; Turner, et. al., 2006; Bazzocchi, et. al., 2008).
Currently, the Wolbachia genomes are being utilized to identify possible targets for
therapeutic agents. Wolbachea can also be used in proliferation of arthropod population.

Another application of the cytoplasmic incapability caused by Wolbachia, is the

proliferation of the arthropod population. Manipulation of Wolbachia-infected males as a
tool to decrease pest population by inducing elevated cytoplasmic incapability, same as
to how the use of sterile male programmes to control pest like insects (Xi, Koo, & Dobson,
2005; Zabalou, et. al., 2004; Dogson, Fox, & Jiggins, 2002). The use of Wolbachia in
resolving problem is not limited with filarial pathogenicity and proliferation of the arthropod
population. It can also be used in shortening the lives of vectors like Aedes mosquitoes
in which the disease agent for dengue fever requires a long incubation time within the
vector (Brownstein, Hett & O’Neill, 2003).
 REFERENCES
Baldo, L., Bordenstein, S., Wernegreen, J. J., & Werren, J. H. (2005). Widespread
recombination throughout Wolbachia genomes. Molecular biology and evolution,
23(2), 437-449.
Bazzocchi, C., Mortarino, M., Grandi, G., Kramer, L. H., Genchi, C., Bandi, C., ... &
McCall, J. W. (2008). Combined ivermectin and doxycycline treatment has
microfilaricidal and adulticidal activity against Dirofilaria immitis in experimentally
infected dogs. International journal for parasitology, 38(12), 1401-1410.
Bourtzis, K. (2008). Wolbachia-based technologies for insect pest population control. In
Transgenesis and the management of vector-borne disease (pp. 104-113).
Springer, New York, NY.
Brownstein, J. S., Hett, E., & O’Neill, S. L. (2003). The potential of virulent Wolbachia to
modulate disease transmission by insects. Journal of invertebrate pathology,
84(1), 24-29.
Debrah, A. Y., Mand, S., Marfo‐Debrekyei, Y., Batsa, L., Pfarr, K., Buttner, M., ... &
Hoerauf, A. (2007). Macrofilaricidal effect of 4 weeks of treatment with
doxycycline on Wuchereria bancrofti. Tropical medicine & international health,
12(12), 1433-1441.
Debrah, A. Y., Mand, S., Specht, S., Marfo-Debrekyei, Y., Batsa, L., Pfarr, K., ... &
Hoerauf, A. (2006). Doxycycline reduces plasma VEGF-C/sVEGFR-3 and
improves pathology in lymphatic filariasis. PLoS pathogens, 2(9), e92.
Dobson, S. L., Fox, C. W., & Jiggins, F. M. (2002). The effect of Wolbachia-induced
cytoplasmic incompatibility on host population size in natural and manipulated
systems. Proceedings of the Royal Society of London. Series B: Biological
Sciences, 269(1490), 437-445.
Kageyama, D., & Traut, W. (2004). Opposite sex–specific effects of Wolbachia and
interference with the sex determination of its host Ostrinia scapulalis.
Proceedings of the Royal Society of London. Series B: Biological Sciences,
271(1536), 251-258.
Lassy, C. W., & Karr, T. L. (1996). Cytological analysis of fertilization and early
embryonic development in incompatible crosses of Drosophila simulans.
Mechanisms of development, 57(1), 47-58.
Narita, S., Kageyama, D., Nomura, M., & Fukatsu, T. (2007). Unexpected mechanism of
symbiont-induced reversal of insect sex: feminizing Wolbachia continuously acts
on the butterfly Eurema hecabe during larval development. Appl. Environ.
Microbiol., 73(13), 4332-4341.
O'Neill, S. L., Giordano, R., Colbert, A. M., Karr, T. L., & Robertson, H. M. (1992). 16S
rRNA phylogenetic analysis of the bacterial endosymbionts associated with
cytoplasmic incompatibility in insects. Proceedings of the National Academy of
Sciences, 89(7), 2699-2702.
Pfarr, K. M., & Hoerauf, A. M. (2006). Antibiotics which target the Wolbachia
endosymbionts of filarial parasites: a new strategy for control of filariasis and
amelioration of pathology. Mini reviews in medicinal chemistry, 6(2), 203-210.
Poinsot, D., Charlat, S., & Mercot, H. (2003). On the mechanism of Wolbachia‐induced
cytoplasmic incompatibility: Confronting the models with the facts. Bioessays,
25(3), 259-265.
Reed, K. M., & Werren, J. H. (1995). Induction of paternal genome loss by the paternal‐
sex‐ratio chromosome and cytoplasmic incompatibility bacteria (Wolbachia): a
comparative study of early embryonic events. Molecular reproduction and
development, 40(4), 408-418.
Richards Jr, F. O., Amann, J., Arana, B., Punkosdy, G., Klein, R., Blanco, C., ... &
Maguire, J. H. (2007). No depletion of Wolbachia from Onchocerca volvulus after
a short course of rifampin and/or azithromycin. The American journal of tropical
medicine and hygiene, 77(5), 878-882.
Ros, V. I. D., Fleming, V. M., Feil, E. J., & Breeuwer, J. A. J. (2009). How Diverse Is the
Genus Wolbachia? Multiple-Gene Sequencing Reveals a Putatively New
Wolbachia Supergroup Recovered from Spider Mites (Acari: Tetranychidae).
Retrieved from https://aem.asm.org/content/75/4/1036
Saint André, A. V., Blackwell, N. M., Hall, L. R., Hoerauf, A., Brattig, N. W., Volkmann,
L., ... & Diaconu, E. (2002). The role of endosymbiotic Wolbachia bacteria in the
pathogenesis of river blindness. Science, 295(5561), 1892-1895.
Sasaki, T., & Ishikawa, H. (2000). Transinfection of Wolbachia in the Mediterranean
flour moth, Ephestia kuehniella, by embryonic microinjection. Heredity, 85(2),
130.
Stouthamer, R., & Kazmer, D. J. (1994). Cytogenetics of microbe-associated
parthenogenesis and its consequences for gene flow in Trichogramma wasps.
Heredity, 73(3), 317.
Turner, J. D., Mand, S., Debrah, A. Y., Muehlfeld, J., Pfarr, K., McGarry, H. F., ... &
Hoerauf, A. (2006). A randomized, double-blind clinical trial of a 3-week course of
doxycycline plus albendazole and ivermectin for the treatment of Wuchereria
bancrofti infection. Clinical Infectious Diseases, 42(8), 1081-1089.
Vandekerckhove, T., Watteyne, S., Bonne, W., Vanacker, D., Devaere, S., Rumes, B.,
... & Mertens, J. (2003). Evolutionary trends in feminization and intersexuality in
woodlice (Crustacea, Isopoda) infected with Wolbachia pipientis (alpha-
Proteobacteria). Belgian journal of zoology, 133(1), 61-69.
Weeks, A. R., & Breeuwer, J. A. J. (2001). Wolbachia–induced parthenogenesis in a
genus of phytophagous mites. Proceedings of the Royal Society of London.
Series B: Biological Sciences, 268(1482), 2245-2251.
Werren, J. H., Baldo, L., & Clark, M. E. (2008). Wolbachia: master manipulators of
invertebrate biology. Nature Reviews Microbiology, 6(10), 741–751.
doi:10.1038/nrmicro1969
World Mosquito Program (n.d.). Wolbachia. Retrieved from
http://www.eliminatedengue.com/our-research/wolbachia
Xi, Z., Khoo, C. C., & Dobson, S. L. (2005). Wolbachia establishment and invasion in an
Aedes aegypti laboratory population. Science, 310(5746), 326-328.
Zabalou, S., Riegler, M., Theodorakopoulou, M., Stauffer, C., Savakis, C., & Bourtzis, K.
(2004). Wolbachia-induced cytoplasmic incompatibility as a means for insect pest
population control. Proceedings of the National Academy of Sciences, 101(42),
15042-15045.