Diel vertical migration of the marine copepod<Emphasis Type="Italic"> Calanopia americana</Emphasis>. I. Twilight DVM and its relationship to the diel light cycle

Marine copepods commonly exhibit vertical movements in the water column over the diel cycle, termed diel vertical migration (DVM), with the most common pattern being an ascent in the water column to minimum depth around sunset and descent to maximum depth around sunrise. The present study characterized the DVM pattern of the pontellid copepod Calanopia americana Dahl in the Newport River estuary (North Carolina, USA, in July 2003). The estuary is shallow and well-mixed, and the study site (34°43N; 76°40W), 1.5 km inside the estuary entrance, is unusual in lying within a gyre where tidal currents are always in the seaward direction. Changes in C. americana vertical abundance were related to spectrally relevant changes in light throughout the diel cycle. Simultaneous measurements of light and zooplankton abundance near the surface (0.5 m depth) and near the bottom (0.5 m above bottom) were made over one 4-h period and two 3-day periods during different phases of the tide. These observations suggest that C. americana undertook twilight DVM in the Newport River estuary; an ascent to the surface occurred at sunset, followed by a descent to near the bottom around midnight, with a second ascent to the surface and then descent to near bottom at sunrise. DVM in C. americana was independent of the tidal cycle, with the initial ascent in the water column at sunset possibly associated with relative rates of irradiance change. Copepod vertical movements were consistent with a night-active endogenous rhythm, and appeared independent of the abundance of predatory chaetognaths, Sagitta spp. In DVM studies with migrators like C. americana that are broadly sensitive to visible wavelengths of light, measuring photosynthetically active radiation may be a reasonable alternative to measuring light in a spectrally relevant photometric unit.Electronic Supplementary Material Supplementary material is available in the online version of this article at http://dx.doi.org/10.1007/s00227-005-1569-x.Communicated by J.P. Grassle, New Brunswick     

Impacts of copepods on marine seston,and resulting effects on<Emphasis Type="Italic"> Calanus finmarchicus</Emphasis> RNA:DNA ratios in mesocosm experiments

We investigated the impact of copepods on the seston community in a mesocosm set-up, and assessed how the changes in food quantity, quality and size affected the condition of the grazers, by measuring the RNA:DNA ratios in different developmental stages of Calanus finmarchicus. Manipulated copepod densities did not affect the particulate carbon concentration in the mesocosms. On the other hand, chlorophyll a content increased with higher copepod densities, and increasing densities had a positive effect on seston food quality in the mesocosms, measured as C:N ratios and 3:6 fatty acid ratios. These food quality indicators were significantly correlated to the nutritional status of C. finmarchicus. In contrast to our expectations, these results suggest a lower copepod growth potential on higher quality food. However, in concordance with earlier studies, we found that when copepods were in high densities the large particles (>1000 µm³) decreased and that the smaller particles (<1000 µm³) increased in number. These patterns were closely linked to the condition of C. finmarchicus, which were of better condition (RNA:DNA ratios) with increasing biovolumes of large particles, and, conversely, lower RNA:DNA ratios with increasing biovolumes of smaller particles. Consequentially, the selective grazing by copepods stimulated increased biovolumes of smaller plankton, and this increase was responsible for the increased food quality, in terms of C:N and 3:6 ratios. Thus, we conclude that the decreasing growth potentials of C. finmarchicus were a result of a decrease of favourably sized food particles, induced by copepod grazing.Communicated by O. Kinne, Oldendorf/Luhe     

Swimming pattern as an indicator of the roles of copepod sensory systems in the recognition of food

The roles of copepod sensory systems in the recognition of food were investigated using the Bugwatcher, a video-computer system designed to track and describe quantitatively the swimming patterns of aquatic organisms. The swimming behavior of the copepodPseudocalanus minutus in the presence of phytoplankton is characterized by a decrease in average swimming speed and an increase in pause behaviors compared to its swimming behavior in filtered seawater. Copepods exposed to chemosensory stimulation alone (filtered phytoplankton exudate) exhibited an increase in average swimming speed and an increase in the number of burst swimming behaviors. When exposed to a novel, non-food chemosensory stimulus (morpholine), no change in swimming behavior was observed unless the copepods had been conditioned to this odor in the presence of phytoplankton. Copepods exposed to mechanosensory stimulation alone (plastic spheres) exhibited a decrease in swimming speed and an increase in pause behaviors. When exposed to both forms of stimulation simultaneously (phytoplankton exudate and plastic spheres), a further decrease in swimming speed and increase in pause behaviors occurs, yielding a swimming pattern similar to that found in the presence of phytoplankton. This analysis of swimming pattern indicates that both chemoreception and mechanoreception contribute to the recognition of food inP. minutus.Contribution No. 406 of the US EPA Environmental Research Laboratory; Narragansett, Rhode Island 02882, USA     

Descriptions and durations of premolt setal stages in female Dungeness crabs,<Emphasis Type="Italic"> Cancer magister</Emphasis>

For crustaceans with a well-defined annual molting season, such as adult female Dungeness crabs (Cancer magister Dana), setal molt staging can, in principle, be used to predict molting destiny of individual crabs. Size-specific predictions of molting probability could, in turn, be useful for estimating mortalities due to molting. Female Dungeness crabs were collected January–March 1997 at depths of 10–30 m off the California coast, USA (41°N). Intermolt stage A₁–C₄ and premolt stages D₀, D₁, D₁, D₁ and D₂ were described based on features of the branchial epipod. Laboratory experiments suggested that stage D₁ was the earliest stage beyond which eventual molting was inevitable so that molting destiny could be determined. Estimated mean number of days from the beginning of stage D₁ to molting was 85. Estimated individual stage durations, based on laboratory experiments, were 24, 51, –4, 18, and 20 days, respectively, for stages D₀, D₁, D₁, D₁, and >D₂. The estimated –4 days for stage D₁ suggests that this stage must be very brief and raises a question of its utility in designation of molt stages. Molt staging of three field samples, 300–600 crabs in each, collected prior to the 1997 annual molting season, indicated an increase in the size and frequency of crabs staged D₁ or later as the time to molt approached, although crabs of 150 mm carapace width and larger showed few signs of molt preparation. Because the duration of the molting season (approximately 120 days) for adult female C. magister in northern California exceeds the estimated maximum duration of reliable prediction of molting destiny (85 days), it does not appear that molt staging can be used to predict molting destiny in this population. However, the procedures that we have used in this paper for application to female C. magister might be used with success for other crustaceans if the duration of stages D₁ to molting exceeds the duration of the molting season, and furthermore may be used for describing temporal molting trends.Communicated by J.P. Grassle, New Brunswick     

Effects of light intensity on nitrate and nitrite uptake and excretion by Chaetoceros curvisetus

Michaelis-Menten uptake kinetics were observed at all light intensities. With constant illumination, the V_max and K₁ in nitrate uptake over the natural light intensity range of 0 to 2000 E were 0.343 g-at NO₃–N(g)^-1 at protein-N h^-1 and 26 E, respectively. Nitrate uptake was inhibited at higher light intensities. The K_s for nitrate uptake did not vary as a function of light intensity remaining relatively constant at 0.62 g-at NO₃–N 1^-1. With intermittent illumination, the V_mzx for light intensity in nitrate uptake over a light intensity range of 0 to 5000 E was 0.341 g-at NO₃–N(g)^-1-at protein-N h^-1. No inhibition of nitrate uptake was observed at higher than natural light intensities. Chaetoceros curvisetus will probably never experience light inhibition of nitrate uptake under natural conditions.     

Primary production in Dumbell Bay in the Arctic Ocean

Photosynthesis, chlorophyll, inorganic nutrients, and related factors were measured throughout a productive season in a small coastal inlet of the Arctic Ocean. Significant production was confined to 5 m for a very limited time period beacause of both light and nutrient limitations. Maximum chlorophyll was 8.2 mg Chl a m^-3 and maximum gross and net photosynthesis rates were 830 and 550 mg C m^-2 d^-1. Annual gross and net photosynthesis is estimated at about 12 and 9 g C m^-2 respectively. The effect of light stimulation on assimilation is described, sources of coastal zone nutrient enrichment are considered, and discrepancies between gross and net photosynthesis are discussed with reference to the growth characteristics of the phytoplankton population.Bigelow Laboratory for Ocean Sciences. Contribution No. 76008     

Triterpenoid and caffeic acid derivatives in the leaves of ragweed, <Emphasis Type="Italic">Ambrosia artemisiifolia</Emphasis> L. (Asterales: Asteraceae), as feeding stimulants of <Emphasis Type="Italic">Ophraella communa</Emphasis> LeSage (Coleoptera: Chrysomelidae)

Summary. The leaf beetle Ophraella communa infests almost exclusively Ambrosia artemisiifolia in the fields of Japan, even though it normally feeds on several Asteraceous plants. A filter paper bioassay showed that the feeding of O. communa is strongly stimulated by methanolic extracts of A. artemisiifolia. The feeding stimulants for O. communa have been isolated from methanolic extracts of A. artemisiifolia. -Amyrin acetate, -amyrin acetate, 5-caffeoylquinic acid (chlorogenic acid) and 3,5-dicaffeoylquinic acid from A. artemisiifolia have been identified as feeding stimulants for O. communa. Triterpenoid derivatives (-amyrin acetate or -amyrin acetate) and caffeic acid derivatives (3, 5-dicaffoylquinic acid or 5-caffeoylquinic acid) showed feeding stimulant activity when mixed together.     

A comparative study of photoadaptation in four diatoms isolated as endosymbionts from larger foraminifera

Four endosymbiotic diatoms were isolated from 2 species of larger foraminifera collected in the Red Sea and Hawaii. The photoadaptive responses of the cultured diatoms were measured at 312, 19 and 7 W cm^-2. Two of the diatoms (Fragilaria shiloi and Nitzschia laevis), both isolated from Amphistegina lessonii, grew fastest at 312 W cm^-2. The other two diatoms (N. valdestriata and N. panduriformis) which were isolated from Heterostegina depressa, grew best at 19 W cm^-2. Of the four diatoms, F. shiloi grew best at high light levels. Also in F. shiloi, chlorophyll c content per cell was directly proportional to light intensity; in contrast chlorophyll a and carotenoids increased to maxima at 19 W cm^-2. The chlorophyll a and c and carotenoid content of N. valdestriata were also maximal at 19 W cm^-2. Photosynthetic rates, measured by respirometry, suggested that the diatoms were photoinhibited at higher light intensities and did well at moderately low light intensities (175W cm^-2). The photocompensation points of all 4 diatoms were about 2% of the light available in the spring at 1-m depth at Elat on the Red Sea. At Elat the photocompensation point would lie between 40 and 50 m if the algae were free in nature. The amount of attenuation of light by the shells of the host has not yet been measured. Presumably photocompensation of the algae within hosts is reached at depths less than 40 m.     

Effect of inhibitors of carbonic anhydrase activity on photosynthesis in the red alga Soliera filiformis (Gigartinales: Rhodophyta)

The effect of light intensity, pH and carbonic anhydrase (CA) inhibitors on photosynthesis of the red marine macroalgae Solieria filiformis (Kützing) Gabrielson, collected from Taliarte (Gran Canaria, Canary Islands) in 1991, has been investigated. Plants taken from the sea (wild phenotype) developed spherical morphology (ball phenotype) after 2 mo culture in aerated tanks. The photosynthetic oxygen evolution in the wild phenotype was saturated at 100 mol photons m^-2s^-1, while the ball phenotype displayed saturation at 200 mol photons m^-2s^-1. The inhibitors of total CA activity (6-ethoxizolamide) and extracellular CA activity (dextran-bound sulfonamide) inhibited photosynthesis at pH 8.2, to 90 and 50% respectively, in both phenotypes. No inhibition of the photosynthetic oxygen evolution was detected at pH 6.5. CA activity was associated with both supernatant and pellet fractions of crude extracts of S. filiformis. The rate of alkalization of the medium by the algae was dependent on light intensity. We suggest that carbon dioxide is the general form of inorganic carbon transported across the plasmamembrane in S. filiformis. HCO₃ transport into the cell takes place simultaneously by an indirect mechanism (dehydration to CO₂ catalyzed by CAext) and by direct uptake. Extracellular (CAext) and intracellular (CAint) CAs are involved in the mechanisms of inorganic carbon assimilation by S. filiformis.     

Ontogenetic diel vertical migration of the planktonic copepodCalanus sinicus in the Inland Sea of Japan

The ontogenetic diel vertical migration of the planktonic copepodCalanus sinicus was investigated in the Inland Sea of Japan in summer 1988, when the water was thermally stratified with a thermocline of ca 5 °C between 35 and 45 m. Stage-specific differences in the diel vertical migration behavior ofC. sinicus were found. Eggs were spawned primarily within the surface-waters between midnight and dawn by ascending females, and sank gradually to deeper waters until they hatched into nauplii. Non-feeding nauplius stages (NI and II) were distributed throughout the water column, but the first feeding stage (NIII) performed an ontogenetic upward migration. NIV to VI and copepodite (C) stages I to III continuously aggregated in the phytoplankton-rich euphotic layer. However, the depth of the median CI to III populations descended as stage progressed. The onset of prominent diel vertical migration took place in CIV, and the amplitude of vertical migration increased with age, being maximal in adult females (CVI). Adult males (CVI), however, remained in the layer below 20 m, and did not migrate dielly. The ecological significance of ontogenetic diel vertical migration is discussed.     

Effect of daylight variations on the energy budgets of shallow-water corals

Energy budgets were determined for small pieces (nubbins) of the coralsPocillopora damicornis, Montipora verrucosa andPorites lobata living at a water depth of 3 m on the fringing reef of Coconut Island, Kaneohe, Hawaii. The budgets were determined for three different types of day: an ideal day with no cloud and an in situ daily integrated irradiance at 3 m of 14.385 E m^–2 d^–1; a normal day with sporadic cloud cover and daily irradiance of 11.915 E m^–2 d^–1; and an overcast day with daily irradiance of 6.128 E m^–2 d^–1. On the ideal day, the energy fixed in photosynthesis was more than that required for respiration and growth of both zooxanthellae and animal components of the association, and there was a predicted loss of between 19.3 and 32.4% of the energy fixed. On a normal day, the total photosynthetic energy fixation was lower and the excess was between 12.1 and 27.9% of the energy fixed. On the overcast day, however, in bothPocillopora damicornis andPorites lobata energy expenditure exceeded photosynthetic energy fixation and the budget was in deficit. Estimates of rate of mucus secretion on an overcast day were derived and, when incorporated into the energy budget, it was predicted that all three species would have a deficit budget, necessitating the catabolism of lipid reserves. From published values for lipid storage in these species it was calculated that the reserves would last from 28 d inPocillopora damicornis to 114 d inM. verrucosa. A model is suggested in which corals draw upon their extensive lipid stores on days of sub-optimal light, replenishing the reserves again when daily light levels are high, and finally excreting the excess energy fixed, as mucus-lipid when the lipid stores are replete.     

Carbon fixation in marine phytoplankton: carboxylase activities and stable carbon-isotope ratios; physiological and paleoclimatological aspects

We measured the activity of three carboxylases: RuBP carboxylase, PEP carboxylase and PEP carboxykinase of marine phytoplankton species in culture and in natural communities. Activities of the three carboxylases were measured simultaneously with stable carbon-isotope ratios. The enzymatic activities have been used to estimate the importance of carboxylation and its impact on the ¹³C:¹²C ratio (expressed as ¹³C). The marine phytoplankton species in culture were Fragilariopsis kerguelensis, Nitzschia turgiduloides, Skeletonema costatum, Phaeodactylum tricornutum, Isochrysis galbana, Dunaliella marina, and Prorocentrum micans, and the field samples were collected from different depths off the coast of Portugal (August/September, 1981). Our results indicate that, as in terrestrial plants, the ¹³C value is a good indicator of the extent of carboxylation. RuBP carboxylase activity was always predominant, whereas the ¹³C value never reached values typical of the C₄ pathway. The carboxylases could be PEP carboxylase (in dinoflagellates) or PEP carboxykinase (in diatoms). carboxylation increased at the end of the exponential growth phase in a diatom culture and with increased biomass in natural samples. We interpret these increases as an adaptative response mechanism to poor environmental conditions, especially to low light intensity.     

Vertical distribution and feeding patterns of midwater fish in the central equatorial Atlantic II. Sternoptychidae

The vertical distribution of seven sternoptychid species was examined from RMT 1+8 samples collected aboard R. V. Meteor in March-April 1979 and from Royal Research Ship R.R.S. Discovery in July 1974 in the central equatorial Atlantic. During daytime sternoptychids occupied depths between 200 and 1250 m, with Sternoptyx pseudobscura living deepest, centering between 800 and 900 m, and Argyropelecus sladeni most shallow, aggregating predominantly at 300 and 400 m. They are all considered limited or partial migrants, ascending only some 100 and 200 m towards the surface at night. Only A. sladeni was observed to enter the epipelagic zone (0 and 200 m).-Feeding patterns were investigated from stomach content analyses of Sternoptyx diaphana, S. pseudobscura, Argyropelecus sladeni and A. affinis. Additional stomach contents were analysed from samples of S. diaphana, A. hemigymnus and A. olfersi collected in June 1985 from F.R.V. Walther Herwig in the temperate NE Atlantic at 46°N, 17°W by means of the Engel Trawl. The food spectrum of the six species is generally described, and additional dietary evidence regarding calanoid copepod prey is provided for four of these taxa. All sternoptychid species investigated were planktivorous, feeding predominantly on copepods and ostracods, except for the largest size class, which preyed heavily on euphausiids and amphipods. The relationship of predator size towards prey type and prey size is analysed for both Sternoptyx species. Of these, S. pseudobscura in particular exhibits taxonomic selectivity towards polychaete prey. The diet of both species of Sternoptyx included a number of epipelagic or even neustonic calanoid copepod species which contributed more than 50% of the total copepod population by numbers. So far it is not known how the predators find access to prey organisms of the upper 200 m, as netfeeding is considered unlikely. Cyclopoid copepods of the genus Sapphirina were observed as dietary component particular of S. diaphana.     

Feeding deterrence properties of apo-fucoxanthinoids from marine diatoms. I. Chemical structures of apo-fucoxanthinoids produced by Phaeodactylum tricornutum

Methanol extracts of freshly harvested cells of the marine diatom Phaeodactylum tricornutum were found to produce a feeding deterrent effect in the copepod Tigriopus californicus. Bioassay guided fractionation of the methanol extracts led to the isolation of four compounds possessing feeding deterrent activity. The compounds were identified as apo-10-fucoxanthinal (1), apo-12-fucoxanthinal (2), apo-12-fucoxanthinal (3), and apo-13-fucoxanthinone (4) by detailed spectroscopic analysis and comparison with authentic compounds produced semi-synthetically from fucoxanthin. Compounds 1 to 4 exhibited feeding deterrent responses in T. californicus at concentrations of less than 20 ppm.     

Further insight on carapid—holothuroid relationships

Carapidae (or pearlfish) are eel-like fishes that live inside different invertebrates, such as holothurians, sea stars, or bivalves. Those of the genus Carapus are commensal and use their host as a shelter, while Encheliophis species are parasitic and eat the hosts gonads. In areas where they live in sympatry, C. boraborensis, C. homei, C. mourlani and E. gracilis are able to inhabit the same host species. Infestation is considered as monospecific when several conspecifics are observed in the same host. However, many aspects of this particular relation remain obscure, e.g. communication between carapids and the defence systems of the different protagonists (carapids and hosts). Experiments have been conducted in the field and laboratory to investigate several aspects of the carapids relationships with their hosts. Sampling carried out in the Bay of Opunohu (Moorea, French Polynesia) determined the sex ratio of C. boraborensis (3:1) and C. homei (1:1) and their distribution rate within different Echinodermata. Our study showed that neither species was capable of determining whether a heterospecific already occupied a sea cucumber or not. They were, however, able to locate the sea cucumbers cloaca, due to the excurrent resulting from respiration. The sea cucumbers defence system (Cuverian tubules) minimises predator attacks, but is not effective against carapid intrusion. The Carapidae defence system is twofold. Due to a passive system related to the sea cucumbers low cloacal position, the Cuverian tubules are not expelled when fish enter the cloaca. Moreover, carapids resist sea cucumber toxins better than other reef fish. Their increased resistance might be related to their gills rather than to their mucus coating; however, the latter may assist the fish in resisting the sticky substances emitted by the Cuverian tubules.Communicated by S.A. Poulet, Roscoff     

Chitinolytic enzymes in the integument and midgut-gland of the shrimpPalaemon serratus during the moulting cycle

Chitinase andN-acetyl--D-glucosaminidase activity were quantified inPalaemon serratus (Pennant) integument and midgut gland during the moulting cycle. Studies were performed on specimens collected near Concarneau, France, in July 1989. The changes in specific activity are different in the two organs: in the midgut gland enzymatic activity is high throughout the whole moulting cycle with a weak peak at the early premoult Stage D1, whereas in the integument the activity of both enzymes is very low throughout post- and intermoult stages and rises only after premoult Stage D1. The highest specific activity is reached in D1 for chitinase and somewhat earlier (D1) forN-acetyl--D-glucosaminidase. The increase in specific chitinolytic activity coincides with an increase in ecdysteroids.     

Effects of nitrogen and phosphorus enrichement on in vivo symbiotic zooxanthellae of Pocillopora damicornis

The reef coral Pocillopora damicornis (Linnaeus) was grown for 8 wk in four nutrient treatments: control, consisting of ambient, unfiltered Kaneohe Bay seawater [dissolved inorganic nitrogen (DIN, 1.0 M) and dissolved inorganic phosphate (DIP, 0.3 M)]; nitrogen enrichment (15 M DIN as ammonium); phosphorus enrichment (1.2 M DIP as inorganic phosphate); and 15 M DIN+1.2 M DIP. Analyses of zooxanthellae for C, N, P and chlorophyll a after the 8 wk experiment indicated that DIN enrichment increased the cellular chlorophyll a and excess nitrogen fraction of the algae, but did not affect C cell^-1. DIP enrichment decreased both C and P cell^-1, but the decrease was proportionally less for C cell^-1. the response of cellular P to both DIN and DIP enrichment appeared to be in the same direction and could not be explained as a primary effect of external nutrient enrichment. The observed response of cellular P might be a consequence of in situ CO₂ limitation. DIN enrichment could increase the CO₂ (aq) demand by increasing the net production per unit area. DIP enrichment could slow down calcification, thus decreasing the availability of CO₂ (aq) in the coral tissue.Hawaii Institute of Marine Biology Contribution No. 920     

Do Tidal-Channel Turbulence Measurements Support k−5/3?

We seriously challenge the belief that tidal-channel data of Grant et al. support the theory for the classical energy-spectrum behavior, k ^–5/3 in the inertial subrange [where k is wave number and (k) is the one-dimensional energy-spectrum function]. The behavior revealed by our re-examination is explained in terms of a more recent theory.     

Effects of anoxia and hypoxia on the dormancy and survival of subitaneous eggs of<Emphasis Type="Italic"> Acartia bifilosa</Emphasis> (Copepoda: Calanoida)

Subitaneous eggs of the copepod Acartia bifilosa were incubated at low oxygen concentrations and in anoxic conditions to study the induction of dormancy and the longevity of the resting eggs. Eggs were incubated in hypoxic conditions for 8 or 10 days, and also in anoxic conditions for 8–441 days. After the anoxic and hypoxic incubations, the unhatched eggs were subsequently incubated in normal oxygen conditions to reveal the viability of the eggs. Low oxygen concentrations (< 0.17 ml l^–1) induced dormancy in the eggs: they did not hatch at these concentrations in the hypox experiments but did hatch afterwards. Eggs survived well in anoxic conditions: after 10 months at 4°C, approximately 40% of the eggs still hatched, while some eggs survived in anoxic conditions for a year at 13°C, which is longer than hitherto reported for subitaneous eggs of any copepod species. Also, sediment that contained eggs of copepods was stored in the laboratory to reveal the long-term survival of the eggs. When incubated after 1.8 years storage, more than 40% of A. bifilosa eggs hatched. The study shows that subitaneous eggs may also be used as a survival strategy on seasonal time scales. This may partly explain why A. bifilosa does not produce diapause eggs in the present study area in the northern Baltic Sea.Communicated by M. Kühl, Helsingør     

Bacteria as food for marine harpacticoid copepods

Feeding experiments were carried out on the harpacticoid copepods Tisbe holothuriae and Paramphiascella vararensis, using various species of marine and brackish water bacteria as food. Short-term experiments with ³H-labelled bacteria are described, as well as experiments covering longer periods of time using non-labelled, dried bacteria. Results show that for both T. holothuriae and P. vararensis, the amount of dry bacteria consumed ranged from 2.06 to 7.07 g copepod^-1 day^-1, indicating a carbon requirement of 1 to 3.5 g copepod^-1 day^-1. The food value of the bacteria strains tested is given in terms of the generation time of the copepods raised on these bacteria.