New Insights into the Nature of Symbiotic Associations in Aphids: Initial Steps Involved in Aphid Gut Infection by Serratia symbiotica Bacteria

ABSTRACT Symbiotic microorganisms are widespread in nature and can play a major role in the ecology and evolution of animals. The aphid-Serratia symbiotica bacterium interaction provides a valuable model to study the mechanisms behind these symbiotic associations. The recent discovery of cultivable S. symbiotica strains with a free-living lifestyle allowed us to simulate their environmental acquisition by aphids to examine the mechanisms involved in this infection pathway. Here, after oral ingestion, we analyzed the infection dynamics of cultivable S. symbiotica during the host’s lifetime using quantitative PCR and fluorescence techniques and determined the immediate fitness consequences of these bacteria on their new host. We further examined the transmission behavior and phylogenetic position of cultivable strains. Our study revealed that cultivable S. symbiotica bacteria are predisposed to establish a symbiotic association with a new aphid host, settling in its gut. We show that cultivable S. symbiotica bacteria colonize the entire aphid digestive tract following infection, after which the bacteria multiply exponentially during aphid development. Our results further reveal that gut colonization by the bacteria induces a fitness cost to their hosts. Nevertheless, it appeared that the bacteria also offer an immediate protection against parasitoids. Interestingly, cultivable S. symbiotica strains seem to be extracellularly transmitted, possibly through the honeydew, while S. symbiotica is generally considered a maternally transmitted bacterium living within the aphid body cavity and bringing some benefits to its hosts, despite its costs. These findings provide new insights into the nature of symbiosis in aphids and the mechanisms underpinning these interactions. IMPORTANCE S. symbiotica is one of the most common symbionts among aphid populations and includes a wide variety of strains whose degree of interdependence on the host may vary considerably. S. symbiotica strains with a free-living capacity have recently been isolated from aphids. By using these strains, we established artificial associations by simulating new bacterial acquisitions involved in aphid gut infections to decipher their infection processes and biological effects on their new hosts. Our results showed the early stages involved in this route of infection. So far, S. symbiotica has been considered a maternally transmitted aphid endosymbiont. Nevertheless, we show that our cultivable S. symbiotica strains occupy and replicate in the aphid gut and seem to be transmitted over generations through an environmental transmission mechanism. Moreover, cultivable S. symbiotica bacteria are both parasites and mutualists given the context, as are many aphid endosymbionts. Our findings give new perception of the associations involved in bacterial mutualism in aphids.

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Studies on the Life Cycle of Some New Zealand Anisakidae (Nematoda)

10.26686/wgtn.16958437 ◽

2021 ◽

Author(s):

◽

Rosemary Jennifer Hurst

Keyword(s):

Life Cycle ◽

New Zealand ◽

Body Cavity ◽

The Body ◽

In Vitro Cultivation ◽

Free Living ◽

Larval Stages ◽

Factors Influencing ◽

Phocarctos Hookeri

<p>The life cycle of Anisakis simplex in New Zealand waters is described from observations on the morphology, distribution and behaviour of free-living and parasitic stages. Comparison with the life cyles of two other anisakids, Phocanema decipiens Myers 1959 and Thynnascaris adunca Rudolphi 1802 shows differences in distribution, degrees of host specificity, the status of invertebrate hosts, the factors influencing infestation levels of teleost hosts, and the location and pathological effects of infestation. Larval stages occurring in intermediate and paratenic hosts were identified by comparison of larval and adult morphometrics. A. simplex larvae were also positively identified by in vitro cultivation through to adults. Some morphometric variations compared to overseas descriptions are apparent. The ventriculus of A. simplex larvae is shorter relative to body length and the intestinal caecum of P. decipiens is longer relative to ventriculus length. Egg and free-living larval stages were obtained from in vitro cultivation of (A. simplex) and collection of eggs from mature adults from definitive hosts (T. adunca). Eggs of P. decipiens were not obtained. Eggs of A. simplex and T. adunca hatch in 8-11 days at 15 [degrees] C. A. simplex eggs hatch in 6 days at a temperature of 22 [degrees] C and did not hatch in 16 days at 10 [degrees] C. Eggs and free-living stage III larvae of A. simplex and T. adunca are similar in morphology with little differentiation of internal structures. Examination of the stomach contents of pelagic fish infested with anisakids indicated that possible intermediate hosts of A. simplex are the euphausiid Nyctiphanes australis and the decapod Munida gregaria. Possible hosts of T. adunca and M. gregaria are a wide variety of smaller zooplanktonic groups, e.g. decapod larvae and copepods. Larvae of A. simplex were found in one of 8850 N. australis; larvae of T. adunca were found in 69 of 3999 chaetognaths (Sagitta spp.) a medusa and a decapod larva. These larvae are morphologically similar to Stage III larvae from teleosts. No anisakids were found in 3956 Euphausia spp., 1147 M. gregaria and 740 prawns. Twenty five T. adunca larvae and adults were found in 818 freshly eaten M. gregaria in teleost stomachs, indicating that this invertebrate may act as a paratenic and a definitive host. Experimental infection of N. australis and M. gregaria with stage II larvae of A. simplex and T. adunca was unsuccessful. The location of anisakid infestation in three pelagic teleost species, Thyrsites atun, Trachurus novaezelandiae and Trachurus declivis is described. A. simplex larvae are found mainly in the body cavity of all species, at the posterior end of the stomach, with less than one percent occurring in the musculature. Distribution of A. simplex larvae does not change with increasing size of the host or increasing total worm burden. Thyrsites atun have a higher proportion of larvae in the stomach wall (8-13%) compared to Trachurus spp. (< 4%). T. adunca larvae are found infrequently in the body cavity of all three species, on the pyloric caeca and in the stomach wall. Adults and larvae of T. adunca are found more commonly in the alimentary canal, indicating that these teleosts are more important as definitive hosts in the life cycle of this anisakid. P. decipiens larvae are found only in Thyrsites atun and occur mainly in the muscles (98.5%). No quantitative pathogenic effects of anisakid infestation on these teleosts hosts were detected. The main factors influencing the infestation of the three teleost species are age of the host, locality and season. Sex of the host and depth (over the continental shelf, 0-250 m) are not important. A. simplex infestation increased with age in all host species examined, and was higher in Trachurus declivis from the southern-most locality, suggesting the existence of at least two distinct populations of this species. Significant differences in infestation of Thyrsites atun with P. decipiens suggests that this anisakid may be more common in southern localities also. The infestation of Thyrsites atun by larval and adult T. adunca in the alimentary canal is most influenced by season and closely related to diet. Nematode samples were obtained from the marine mammals Arctocephalus forsteri, Kogia breviceps and Phocarctos hookeri. Adult A. simplex were recorded from A. forsteri (a new host record) and Kogia breviceps; preadults from Phocarctos hookeri. Adult P. decipiens were recorded from Phocarctos hookeri; preadults from Arctocephalus forsteri and K. breviceps. Other anisakids found were Anisakis physeteris (Baylis 1923), Contracaecum osculatum Rudolphi 1802 and Pseudoterranova kogiae (Johnston and Mawson 1939) Mosgovoi 1951. These records are all new for the New Zealand region except P. decipiens from P. hookeri and C. osculatum from Arctocephalus forsteri. A. simplex and C. osculatum were found associated with gastric ulcers in Arctocephalus forsteri.</p>

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Studies on the Life Cycle of Some New Zealand Anisakidae (Nematoda)

10.26686/wgtn.16958437.v1 ◽

2021 ◽

Author(s):

◽

Rosemary Jennifer Hurst

Keyword(s):

Life Cycle ◽

New Zealand ◽

Body Cavity ◽

The Body ◽

In Vitro Cultivation ◽

Free Living ◽

Larval Stages ◽

Factors Influencing ◽

Phocarctos Hookeri

<p>The life cycle of Anisakis simplex in New Zealand waters is described from observations on the morphology, distribution and behaviour of free-living and parasitic stages. Comparison with the life cyles of two other anisakids, Phocanema decipiens Myers 1959 and Thynnascaris adunca Rudolphi 1802 shows differences in distribution, degrees of host specificity, the status of invertebrate hosts, the factors influencing infestation levels of teleost hosts, and the location and pathological effects of infestation. Larval stages occurring in intermediate and paratenic hosts were identified by comparison of larval and adult morphometrics. A. simplex larvae were also positively identified by in vitro cultivation through to adults. Some morphometric variations compared to overseas descriptions are apparent. The ventriculus of A. simplex larvae is shorter relative to body length and the intestinal caecum of P. decipiens is longer relative to ventriculus length. Egg and free-living larval stages were obtained from in vitro cultivation of (A. simplex) and collection of eggs from mature adults from definitive hosts (T. adunca). Eggs of P. decipiens were not obtained. Eggs of A. simplex and T. adunca hatch in 8-11 days at 15 [degrees] C. A. simplex eggs hatch in 6 days at a temperature of 22 [degrees] C and did not hatch in 16 days at 10 [degrees] C. Eggs and free-living stage III larvae of A. simplex and T. adunca are similar in morphology with little differentiation of internal structures. Examination of the stomach contents of pelagic fish infested with anisakids indicated that possible intermediate hosts of A. simplex are the euphausiid Nyctiphanes australis and the decapod Munida gregaria. Possible hosts of T. adunca and M. gregaria are a wide variety of smaller zooplanktonic groups, e.g. decapod larvae and copepods. Larvae of A. simplex were found in one of 8850 N. australis; larvae of T. adunca were found in 69 of 3999 chaetognaths (Sagitta spp.) a medusa and a decapod larva. These larvae are morphologically similar to Stage III larvae from teleosts. No anisakids were found in 3956 Euphausia spp., 1147 M. gregaria and 740 prawns. Twenty five T. adunca larvae and adults were found in 818 freshly eaten M. gregaria in teleost stomachs, indicating that this invertebrate may act as a paratenic and a definitive host. Experimental infection of N. australis and M. gregaria with stage II larvae of A. simplex and T. adunca was unsuccessful. The location of anisakid infestation in three pelagic teleost species, Thyrsites atun, Trachurus novaezelandiae and Trachurus declivis is described. A. simplex larvae are found mainly in the body cavity of all species, at the posterior end of the stomach, with less than one percent occurring in the musculature. Distribution of A. simplex larvae does not change with increasing size of the host or increasing total worm burden. Thyrsites atun have a higher proportion of larvae in the stomach wall (8-13%) compared to Trachurus spp. (< 4%). T. adunca larvae are found infrequently in the body cavity of all three species, on the pyloric caeca and in the stomach wall. Adults and larvae of T. adunca are found more commonly in the alimentary canal, indicating that these teleosts are more important as definitive hosts in the life cycle of this anisakid. P. decipiens larvae are found only in Thyrsites atun and occur mainly in the muscles (98.5%). No quantitative pathogenic effects of anisakid infestation on these teleosts hosts were detected. The main factors influencing the infestation of the three teleost species are age of the host, locality and season. Sex of the host and depth (over the continental shelf, 0-250 m) are not important. A. simplex infestation increased with age in all host species examined, and was higher in Trachurus declivis from the southern-most locality, suggesting the existence of at least two distinct populations of this species. Significant differences in infestation of Thyrsites atun with P. decipiens suggests that this anisakid may be more common in southern localities also. The infestation of Thyrsites atun by larval and adult T. adunca in the alimentary canal is most influenced by season and closely related to diet. Nematode samples were obtained from the marine mammals Arctocephalus forsteri, Kogia breviceps and Phocarctos hookeri. Adult A. simplex were recorded from A. forsteri (a new host record) and Kogia breviceps; preadults from Phocarctos hookeri. Adult P. decipiens were recorded from Phocarctos hookeri; preadults from Arctocephalus forsteri and K. breviceps. Other anisakids found were Anisakis physeteris (Baylis 1923), Contracaecum osculatum Rudolphi 1802 and Pseudoterranova kogiae (Johnston and Mawson 1939) Mosgovoi 1951. These records are all new for the New Zealand region except P. decipiens from P. hookeri and C. osculatum from Arctocephalus forsteri. A. simplex and C. osculatum were found associated with gastric ulcers in Arctocephalus forsteri.</p>

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Survival, feeding and embryo development of Gyrodactylus gasterostei (Monogenea: Gyrodactylidae)

Parasitology ◽

10.1017/s0031182001008861 ◽

2002 ◽

Vol 124 (1) ◽

pp. 53-68 ◽

Cited By ~ 20

Author(s):

J. CABLE ◽

R. C. TINSLEY ◽

P. D. HARRIS

Keyword(s):

Close Contact ◽

Free Living ◽

New Host ◽

Source Of Infection ◽

Parental Feeding ◽

Age Dependent ◽

Reproductive Rates ◽

Dense Vesicles ◽

Tissue Ultrastructure

Viviparous gyrodactylid monogeneans which are detached from their host may form an important source of infection in fish communities. This is particularly significant for these ectoparasites which have no specific free-living stage and where transmission usually occurs through transfer of adult parasites when fish come into close contact. In this study, in vitro survival of Gyrodactylus gasterostei was correlated with changes in tissue ultrastructure of parents and their embryos during detachment and following return to a host. At 10 °C, detached worms survived for up to 89 h (50% survival = 20 h) with mortality influenced by both age-independent and age-dependent factors. The gastrodermis of attached, fed parasites is syncytial and contains a variety of vesicles and feeding vacuoles. However, following detachment and starvation, degenerative changes occur within the gastrodermis, eventually leading to marked damage to the embryo/s. When starved worms reattached to a fish, they began browsing on host epithelial cells within 1 min and ultrastructural evidence for phagocytic activity in the gastrodermis was detected after 5 min. Putative waste vacuoles increased in abundance after 5–30 min on the new host, coinciding with the reappearance of electron-dense vesicles and possibly indicating completion of the first intracellular cycle of digestion. Parental feeding directly influenced the normal cyclical maturation and regression of the uterine lining between births. In extreme cases, starvation led to detached parasites (6·1%) aborting their embryos. However, even short periods off the host influenced development and survival of embryos, suggesting that temporary interruption of nutrient flow to the embryo can significantly affect gyrodactylid reproductive rates.

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Immunological tests by anti-free-living amoebas serum produced in experimental animals I. Immobilization of free-living amoebas in vitro by rabbit antiserum

The Korean Journal of Parasitology ◽

10.3347/kjp.1978.16.1.41 ◽

1978 ◽

Vol 16 (1) ◽

pp. 41 ◽

Cited By ~ 4

Author(s):

Kyung Il Im ◽

Hyea Sook Oh

Keyword(s):

Rabbit Antiserum ◽

Free Living ◽

Experimental Animals ◽

Immunological Tests

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Development of an in vitro culture method for harvesting the free-living infective larvae of Strongyloides venezuelensis

The Korean Journal of Parasitology ◽

10.3347/kjp.1998.36.1.15 ◽

1998 ◽

Vol 36 (1) ◽

pp. 15 ◽

Cited By ~ 9

Author(s):

B K Baek ◽

M K Islam ◽

J H Kim

Keyword(s):

In Vitro Culture ◽

Culture Method ◽

Free Living ◽

Infective Larvae ◽

Strongyloides Venezuelensis

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Current Challenges in the Development of Vaccines and Drugs Against Emerging Vector-borne Diseases

Current Medicinal Chemistry ◽

10.2174/0929867325666181105121146 ◽

2019 ◽

Vol 26 (16) ◽

pp. 2974-2986 ◽

Cited By ~ 4

Author(s):

Kwang-sun Kim

Keyword(s):

New Host ◽

In Vivo Models ◽

Vector Borne Diseases ◽

Novel Drugs ◽

Huge Impact ◽

Tick Borne Disease ◽

Vector Borne ◽

Living Organisms

Vectors are living organisms that transmit infectious diseases from an infected animal to humans or another animal. Biological vectors such as mosquitoes, ticks, and sand flies carry pathogens that multiply within their bodies prior to delivery to a new host. The increased prevalence of Vector-Borne Diseases (VBDs) such as Aedes-borne dengue, Chikungunya (CHIKV), Zika (ZIKV), malaria, Tick-Borne Disease (TBD), and scrub typhus has a huge impact on the health of both humans and livestock worldwide. In particular, zoonotic diseases transmitted by mosquitoes and ticks place a considerable burden on public health. Vaccines, drugs, and vector control methods have been developed to prevent and treat VBDs and have prevented millions of deaths. However, development of such strategies is falling behind the rapid emergence of VBDs. Therefore, a comprehensive approach to fighting VBDs must be considered immediately. In this review, I focus on the challenges posed by emerging outbreaks of VBDs and discuss available drugs and vaccines designed to overcome this burden. Research into promising drugs needs to be upgraded and fast-tracked, and novel drugs or vaccines being tested in in vitro and in vivo models need to be moved into human clinical trials. Active preventive tactics, as well as new and upgraded diagnostics, surveillance, treatments, and vaccination strategies, need to be monitored constantly if we are to manage VBDs of medical importance.

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Effect of Gaseous Ozone on Listeria monocytogenes Planktonic Cells and Biofilm: An In Vitro Study

Foods ◽

10.3390/foods10071484 ◽

2021 ◽

Vol 10 (7) ◽

pp. 1484

Author(s):

Felice Panebianco ◽

Selene Rubiola ◽

Francesco Chiesa ◽

Tiziana Civera ◽

Pierluigi Aldo Di Ciccio

Keyword(s):

Listeria Monocytogenes ◽

In Vitro Study ◽

Planktonic Cells ◽

Free Living ◽

Biofilm Inhibition ◽

Additional Tool ◽

Complete Inactivation ◽

Gaseous Ozone ◽

Food Borne

Among food-borne pathogens, Listeria monocytogenes continues to pose concerns to food business operators due to its capacity to form biofilm in processing environments. Ozone may be an eco-friendly technology to control microbial contaminations, but data concerning its effect on Listeria monocytogenes biofilm are still limited. In this study, the effect of gaseous ozone at 50 ppm on planktonic cells and biofilm of reference and food-related Listeria monocytogenes strains was evaluated. Ozone caused a reduction in microbial loads of 3.7 ± 0.4 and 3.9 ± 0.4 Log10 CFU/mL after 10 and 30 min, respectively. A complete inactivation of planktonic cells after 6 h of treatment was observed. Biofilm inhibition and eradication treatments (50 ppm, 6 h) resulted in a significant decrease of the biofilm biomass for 59% of the strains tested, whilst a slight dampening of live cell loads in the biofilm state was observed. In conclusion, gaseous ozone is not sufficient to completely counteract Listeria monocytogenes biofilm, but it may be useful as an additional tool to contrast Listeria monocytogenes free-living cells and to improve the existing sanitization procedures in food processing environments.

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Assessing the Phytosanitary Risk Posed by an Intraspecific Invasion of Cryphonectria parasitica in Europe

Phytopathology ◽

10.1094/phyto-06-19-0197-r ◽

2019 ◽

Vol 109 (12) ◽

pp. 2055-2063 ◽

Cited By ~ 3

Author(s):

Francesca Dennert ◽

Joana Beatrice Meyer ◽

Daniel Rigling ◽

Simone Prospero

Keyword(s):

United States ◽

South Korea ◽

Plant Pathogens ◽

Castanea Sativa ◽

The United States ◽

Cryphonectria Parasitica ◽

Seedling Mortality ◽

New Host ◽

Phytosanitary Measures

Intraspecific cryptic invasions may occur when new strains of an invasive species are introduced into an area where this species had already been introduced previously. In plant pathogens, such invasions are not well studied, even if, potentially, they can have severe consequences. Here, we investigated the effects of a potential intraspecific invasion in Europe of Cryphonectria parasitica, the causal agent of chestnut blight. Specifically, we tested the hypotheses that (i) non-European strains are more virulent on Castanea sativa than those already present in Europe because they have never encountered this new host, and (ii) the variation in virulence among strains is higher within native than within introduced populations. In a greenhouse, 2-year-old C. sativa seedlings were inoculated with Cryphonectria parasitica strains from South Korea, the United States, and Switzerland, and lesion development and seedling mortality were recorded weekly. Additionally, growth and sporulation of the strains were measured in vitro on agar medium at 15 and 24°C. Although lesion growth was similar for all strains, seedlings inoculated with strains from South Korea and Switzerland died faster than seedlings inoculated with strains from the United States. Moreover, in vitro strains from South Korea grew faster and produced more spores at both temperatures than the strains from the other two countries. In conclusion, our results did not support the two hypotheses. All strains, regardless of their origin, were found to be highly virulent on the inoculated chestnut seedlings. Nevertheless, current phytosanitary measures to avoid the introduction of new genotypes of C. parasitica into Europe should be further implemented.

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