LABORATORY STUDIES ON THE CHEMO-SENSORY REACTIONS OF BLOWFLIES

Attraction of various blowflies to sheep wool has been demonstrated under laboratory conditions using a choice-chamber technique. The attractive factor in wool is intimately associated with the wool fibre for it does not disappear during storage and it is not completely removed by washing.
Only female flies were attracted to wool. The highest degree of attraction was obtained from fertilized females which had daily access to meat. Even in the absence of fertilization and meat, attraction was not completely abolished.
The species and strains tested can be placed in the following order in terms of strength of response to sheep wool: Lucilia sericata (British), L. cuprina (tested on damp floor)>L. sericata (Danish: country)> L. sericata (Danish: city), L. sericata (Australian) > L. illustris, L. caesar. No response was given by Calliphora vomitoria.
The olfactory organs involved in the response to wool are mainly on the antennae but antennaeless L. sericata and L. cuprina with some tarsi removed were still able to show some response to wool provided that they were tested on a damp floor. In the course of testing the olfactory responses it was also shown that the main humidity receptors are present on the antennae.
The existence of a 'wool-factor' to which only certain strains of L. sericata and L. cuprina are specially sensitive offers a possible explanation for the greater efficiency of these species as myiasis-producing forms.     

Orientation of agents of wound myiasis to hosts and artificial stimuli in Hungary

Abstract. The orientation of adults of the myiasis species Wohlfahrtia magnified and Lucilia sericata to artificial visual stimuli (cloth targets) and olfactory stimuli ('swormlure-4', a potent screwworm attractant), was studied in sheep pastures near to Sarbogard in Hungary. Experiments with odour-baited cloth targets, enclosed in electrocuting grids, demonstrated that colour was an important factor influencing catches on targets: a black target caught most flies of both species, with other colours in the following order of effectiveness, blue > white > yellow. Wohlfahrtia magnified did not respond to swormlure-4 in the strong manner that L. sericata did. The sex ratios of W. magnifica caught on targets (67.2% males) and hand-netted from fence posts (68.8% males) were similar and biased towards males, whereas that of L. sericata on targets was strongly biased towards females (15.6% males), indicating a fundamental difference in the response of these two myiasis species towards the swormlure-baited targets. The orientation of these two species towards hosts was also recorded together with that of a third species, Phormia regina . Electric nets placed beside infested sheep caught significantly more flies of all three species than those placed beside uninfested sheep or in the absence of sheep. The sex ratio of W. magnified caught around infested sheep was the reverse of that on targets, with 68.5% females. Wohlfahrtia magnifica responded more strongly to healthy, uninfested sheep than did L. sericata and P. regina . The potential for use of targets both for population monitoring and control is discussed.     

Seasonal and spatial changes in blowfly production from small and large carcasses at Durham in lowland northeast England

Colonization by blowflies (Diptera: Calliphoridae) of mouse carcasses exposed in open agricultural land near Durham (54 45'N) changed from early spring monopolization by Calliphora vicina R.-D. to a summer pattern of multiple species exploitation by this species together with Lucilia caesar L., L. illustris Mg., L. silvarum Mg., L. sericata Mg. and L. richardsi Collin. In a garden at the edge of Durham, mouse carcasses were dominated by C. vicina from spring to autumn. Difference in mouse colonization between the agricultural and garden sites seemed to reflect differences in the blowfly species present, as measured by baited trap catches at the sites. In sets of C. vicina reared from mice under conditions of competition for larval food, it was found that resulting females were significantly larger than males, size being measured as mean wing length. Blowfly production from three sheep carcasses exposed successively at the agricultural site was dominated by C. vomitoria L. and L. caesar, but also produced other Lucilia species in small numbers, including L. sericata. These L. sericata females from sheep that had died from causes other than myiasis included full-sized specimens, in contrast to those produced from mouse carcasses that were all undersized individuals. As L. sericata females trapped on sheep pastures are predominantly full-sized, this suggests that large carcasses may, in part, be a source of the L. sericata population that attacks sheep as a myiasis agent. The nature of large carcasses as possible sources of L. sericata in lowland Britain is discussed.     

STUDIES ON LUCILIA SPECIES (DIPTERA) UNDER DANISH CONDITIONS

Whilst sheep myiasis caused by Lucilia sericata is a severe economic problem in Great Britain, it rarely reaches such proportions in countries bordering Western Europe. Possible reasons for this relative absence of myiasis were investigated in Denmark.
During the period 19 July-15 August 1948, trapping studies showed that blowflies normally associated with myiasis in Great Britain, and particularly L. sericata , occurred fairly abundantly in the Mols area of Jutland.
The reactions of Danish L. sericata to attractive materials placed on sheep were similar to those shown by the species under British conditions. The number of egg batches laid on sheep in response to chemical stimuli were fewer than would have been expected in Great Britain.
Studies on the fleece microclimate showed that relative humidities of 50 % or more frequently occurred on the skin surface. Humidities 3 cm. above the skin were markedly lower. Except during or immediately after rain, humidities high enough to ensure the hatching of blowfly eggs were never recorded.
It is concluded that the absence of myiasis in Denmark and its relatively low incidence in other countries is not due to the existence on the Continent of a strain of L. sericata differing in oviposition habits from the British species.     

Fly abundance and climate as determinants of sheep blowfly strike incidence in southwest England

The relationships between abundance of the blowfly Lucilia sericata (Meigen) (Diptera: Calliphoridae), climate, animal management procedures and the incidence of cutaneous myiasis (blowfly strike) in sheep were examined in three sheep pasture systems in southwest England during the summers of 2002 and 2003. In each year, flies were collected using liver-baited sticky targets, daily weather and routine husbandry practices were noted and the age-class of each animal infested and body position of each strike were recorded. On sites where no strike control was used, 5.8-12.1% of ewes and 5.7-15.8% of lambs were struck. Ewe strikes predominated at the beginning of the season. The incidence of ewe strikes was significantly associated with higher mean temperature, rainfall and shearing; shearing was associated with a 95% reduction in the risk of ewe strike. In lambs, the incidence of strike was significantly related to higher fly abundance, ewe shearing, treatment and mean ambient temperature. Lambs were 4.6 times more likely to be struck after the ewes had been shorn than before; however, the strongest relationship was with mean L. sericata abundance. Average minimum threshold temperatures of 9.5 degrees C for lamb strikes and 8.5 degrees C for all strikes were extrapolated, below which oviposition did not occur. Over 80% of ewe strikes occurred in the breech region in 2002, as did 100% in 2003. However, in lambs both body and breech strikes occurred in both years. The distribution of lamb strikes appeared to change over time, with breech strikes predominating in May, June and July and body strikes occurring increasingly later in the season. The incidence of lamb breech strikes was significantly associated with higher L. sericata abundance and ewe shearing but there was no relationship with weather conditions. By contrast, the risk of body strike in lambs was significantly associated with higher blowfly abundance, higher rainfall and higher maximum temperatures. The relationship between strike incidence and L. sericata abundance is important because it enhances our understanding of strike incidence patterns and management of this disease. Clearly, any factors that facilitate larger L. sericata populations, such as inappropriate carcass burial or increased average ambient temperatures, are likely to increase the incidence of strike.     

Development and laboratory evaluation of chemically‐based baited ovitrap for the monitoring of Aedes aegypti

Aedes (Stegomyia) aegypti is considered to be the most important dengue vector worldwide. Studies were conducted to design and evaluate a chemically‐based baited ovitrap for monitoring Ae. aegypti under laboratory conditions. Several known chemical attractants and three types of ovitraps (ovitraps A, B, and C) were evaluated throughout the oviposition bioassays. Oviposition responses of gravid female Ae. aegypti were evaluated to n‐heneicosane, 3‐methylindole (skatole), 4‐methylphenol (p‐cresol), and phenol. Female Ae. aegypti were attracted to all the evaluated compounds. Among them, n‐heneicosane at a concentration of 10 ppm (mg/l), skatole from 50 to 1000 ppm, p‐cresol at 100 ppm, and phenol at 50 ppm showed a significant positive oviposition response. A blend of the four chemical attractants increased the oviposition response; 67% of the eggs were deposited in the treatment compared to the control. Female Ae. aegypti were signi?cantly more attracted to ovitrap A loaded with the four‐component synthetic blend compared to the standard ovitrap in the oviposition bioassays. The compound used in ovitrap A retained its attractant property for up to three days. The chemically‐based baited ovitrap may be considered as an option to be integrated during the monitoring of dengue virus vectors in México.     

Effects of an artificial blend of host‐infested plant volatiles on plant attractiveness to parasitic wasps

For the biological control of diamondback moth (DBM) larvae in commercial greenhouses, we have previously identified a blend of volatiles that attracted Cotesia vestalis, a parasitoid of DBM larvae. Here, we tested the effects of an artificial volatile blend on the attractiveness of komatsuna plants (Japanese mustard spinach; Brassica rapa var. perviridis) to C. vestalis under greenhouse conditions. First, we showed that female C. vestalis preferred infested komatsuna plants to uninfested plants in the greenhouse. Under the same conditions, placing the artificial attractants near both infested and uninfested plants did not affect the wasps’ preference. However, when comparing infested komatsuna plants coupled with the artificial attractants with infested plants without them, significantly more female C. vestalis were attracted to the former. The possible use of artificial C. vestalis attractants for the biological control of DBM is discussed.     

The use of carrion as breeding sites by the blowfly Lucilia sericata and other Calliphoridae

Abstract. To examine the species composition of flies breeding in carrion in the field, the carcasses of mice and quail were exposed on sheep farms in the South West of England. Calliphora vicina was the dominant species of Diptera; 19,294 individuals emerged from 175 of the 241 infested carcasses recovered. Lucilia sericata emerged from only 39 of the infested carcasses, at a median of 10 adults per infested carcass. Other species of Lucilia present were L.ampullacea, L.caesar and L.illustris. The highest number of L.sericata emerged from carcasses placed in open pasture, the highest number of C.vicina emerged from carcasses in hedgerow, whereas the highest numbers of L.caesar, L.ampullacea and L.illustris emerged from carcasses in woodland. The duration of exposure of carcasses in the field was negatively related to the size of the adult L.sericata which emerged and, in woodland and hedgerow habitats, to the number of L.sericata which emerged. These data indicate that the larvae of L.sericata in carcasses experience significant levels of competition and that the intensity of this competition may be sufficient to reduce the numbers of L.sericata able to emerge successfully. The size distributions of female L. sericata which emerged from carcasses or which were caught as adults in the field showed only a small degree of overlap, suggesting that only a relatively small proportion of the wild L.sericata population emerge from carcass breeding sites. The results are discussed in relation to the development of new approaches to the control of blowfly strike of sheep.     

Adult mortality and oviposition rates in field and captive populations of the blowfly Lucilia sericata

Abstract.  1. Adult mortality and oviposition rates were determined for populations of the blowfly Lucilia sericata (Meigen) (Diptera: Calliphoridae). This species is of economic importance as the primary agent of sheep myiasis throughout north-western Europe.
2. Populations of marked flies in six, 1 m³, outdoor field cages and unmarked wild flies at two farms in south-west England were studied simultaneously between May and September 1998.
3. In the field, wild female L. sericata were caught and aged using a combination of wing-fray and ovarian dissection techniques. Survivorship analysis gave estimates of mortality of 1.94% (± 0.037) and 2.09% (± 0.044) per day-degree and mean life expectancy of 51.5 and 47.9 day-degrees above a threshold of 11 °C, at the two farms studied. Mean lifetime reproductive output in the field was estimated to be 159.6 and 138.4 eggs per female at the two farms respectively.
4. The survivorship of cohorts of marked female flies in cages was followed by counting the number of dead individuals each day; the mortality rate of these flies was 0.81% per day-degree (± 3.49 × 10⁻⁴%) and the mean life expectancy was 123.1 day-degrees above a threshold of 11 °C. Mortality rate was shown to increase significantly with average ambient temperature and relative humidity lagged for two sample periods (approximately 10 days). Oviposition rate also increased with average temperature but declined with average relative humidity. A best-fit multiple regression model incorporating both ambient temperature and humidity explained 60.5% of the variance in the pattern of oviposition.
5. The differences between the field and cage populations highlight the caution required when extrapolating life-history parameters from artificial to natural habitats.     

Development of an attractive target for the sheep blowfly Lucilia sericata

Coloured, odour-baited, sticky targets, designed to catch the sheep blowfly Lucilia sericata Meigen are described and the results of field trials are presented. The number of L. sericata caught by the targets was positively related to target surface area and was dependent on target colour. The performance of the coloured targets is described by a model in which catch is related negatively to reflectivity in the 300-450 nm (ultra-violet/blue) band of the spectrum and positively to the reflectivity in the 450-580 nm (blue-green-yellow) band. Of the Lucilia blowflies caught by the targets, 13-16% were L.sericata and 72-75% of the L.sericata were female. The number of L.sericata caught appeared to be positively related to the availability of sheep over a relatively localized area and showed a marked decline following insecticidal dipping of sheep. The further development of targets and the implications of the results for L.sericata control are discussed.     

Laboratory and field evaluation of an oviposition trap for Culex quinquefasciatus (Diptera: Culicidae)

An ovitrap (BR-OVT) based on physical and chemical stimuli for attracting gravid Culex quinquefasciatus (Diptera: Culicidae) females was developed and evaluated under laboratory and field conditions. Attractants were assayed using alternative chamber bioassays prior to being used in the BR-OVT oviposition trap. A significant preference of gravid females for sites containing conspecific egg rafts was observed, as a response to the natural oviposition pheromone, as well as for sites treated with the synthetic pheromone erythro-6-acetoxy-5-hexadecanolide. Five- to 20-day old grass infusion was strongly attractive to gravid females for laying eggs. On the other hand, entomopathogenic Bacillus sphaericus (Bs) did not influence the choice of an oviposition site when used in combination with grass infusion and can therefore be used as a larvicide in ovitraps. Results from field trials showed that the BR-OVT with grass infusion and with or without Bs works as a preferred oviposition site for Cx. quinquefasciatus. The BR-OVT was more effective for egg collection when placed indoors and comparison with the number of egg rafts laid in cesspits over 40 days indicates that this very simple ovitrap may be a useful tool for monitoring populations of the most important of the vectors of bancroftian filariasis.     

Bimodal cue complex signifies suitable oviposition sites to gravid females of the common green bottle fly

Gravid females of the common green bottle fly, Lucilia sericata Meigen (Diptera: Calliphoridae), readily locate recently deceased vertebrates as oviposition sites, particularly when these animals have been injured. We investigated semiochemical and visual cues that mediate attraction of gravid females to fresh rat carrion. Female flies were more strongly attracted to incised rat carrion than to intact carrion. They were also attracted to Porapak Q headspace volatile (HSV) extract of incised rat carrion. Analyzing aliquots of Porapak Q HSV extract by gas chromatographic‐electroantennographic detection revealed nine components [phenol, para‐ and/or meta‐cresol (could not be separated), guaiacol, dimethyl trisulfide (DMTS), phenylacetaldehyde, (E)‐2‐octenal, nonanal, and tetramethyl pyrazine] that consistently elicited responses from blow fly antennae. In laboratory experiments, a synthetic blend of these nine components was as attractive to gravid females as Porapak Q HSV extract, but blend attractiveness was due entirely to DMTS. In both laboratory and field experiments, increasing doses of DMTS attracted increasingly more flies. Coupled with DMTS, carrion‐type color cues (dark red, black) were more effective than bright color cues (white, yellow) in attracting flies. In field experiments, dark traps baited with DMTS captured a total of 214 calliphorid flies (200 L. sericata, 10 Lucilia illustris Meigen, three Calliphora vicina Robineau‐Desvoidy, one Calliphora vomitoria L.), all of which were gravid females. These results support the conclusion that DMTS and dark color represent a bimodal cue complex that signifies suitable oviposition sites to gravid calliphorid females, particularly L. sericata.     

Comparative field studies of various traps and attractants of the olive fruit fly,Bactrocera oleae

In the framework of an ongoing testing and refining process of a mass trapping method, using a combination of semiochemicals, for the control of the olive fruit fly,Bactrocera (Dacus) oleae, four trap designs, three trap colors, six different food attractants, and two pheromone formulations were compared under field conditions. No differences were observed between the trap designs and the food attractants tested. Trap color had a significant effect in preference tests only. Traps combining food attractants and pheromones attracted higher numbers of both male and female flies but the differences became statistically significant only in cases of low trap densities. Pheromones enclosed in cyclodextrenes were not as attractive as standard pheromone formulations. The findings of these tests allow a choice among trap types and semiochemicals for cost reduction and convenience, but not for enhancement of the efficacy of the method.     

Lifetime reproductive output of Calliphora vicina and Lucilia sericata in outdoor caged and field populations; flight vs. egg production?

Females of the blowflies Calliphora vicina (Robineau-Desvoidy) and Lucilia sericata (Meigen) (Diptera: Calliphoridae) maintained in sheltered outdoor cages and supplied with excess food, oviposited later than would have been expected from the temperature-sum. The survival rates of the caged flies was high and the isolation of flies from predation, extreme temperatures and food shortages is likely to have contributed to this. Despite good survival rates, subsequent egg production over the greater part of the adult life span was reduced to approximately 24% for C. vicina and approximately 55% for L. sericata, of the potential expected from the published temperature-sums required for the maturation of successive egg batches. The data suggest that under field-cage conditions there is a considerable variation in egg development between individuals of the same age and that this variation should not be overlooked, since it may have significant implications in ecological and forensic investigations; however, the cause of this variability remains unclear. While lower than expected, the reproductive outputs recorded in the cages were nevertheless considerably greater than those that have been estimated for blowflies in the field and illustrate the potential for population increase in these species under favourable conditions. The possibility of a greater energy investment in flight activity relative to reproductive output in C. vicina compared to L. sericata is proposed.     

Responses of the sheep blowflies Lucilia sericata and Lcuprina to odour and the development of semiochemical baits

Abstract. The literature relating to the attraction of the sheep blowflies Lucilia sericata and Lucilia cuprina to their ovine hosts is reviewed. The responses of the two species are similar and different components of the behaviour leading to host location and oviposition appear to involve at least two distinct sets of semiochemical cues. Activation, upwind orientation and landing appear to occur in response to putrefactive sulphur-rich volatiles, originating from bacterial decomposition products. Oviposition is elicited primarily by the presence of ammonia-rich compounds; moisture, pheromones and tactile stimuli may also act as oviposition stimuli. There is a pronounced sex difference in the response of Lucilia to semiochemicals with a higher proportion of females attracted than males and a higher proportion of gravid than non-gravid females. While the mechanisms of host location by Lucilia are of intrinsic interest, understanding the responses to semiochemicals is important in the attempt to develop powerful synthetic baits for deployment with the traps or targets used for population sampling or suppression. The literature is discussed with respect to the development of synthetic semiochemical baits.     

Field evaluation of the response of Aedes albopictus (Stegomyia albopicta) to three oviposition attractants and different ovitrap placements using black and clear autocidal ovitraps in a rural area of Same,Timor‐Leste

Known oviposition attractants or stimulants were compared, singly and in combination, using inexpensive autocidal ovitraps designed to trap emerging adults, in a rural area of Timor‐Leste during the dry season. In this area, the dengue vector Aedes albopictus (Stegomyia albopicta) Skuse (Diptera: Culicidae) was abundant, but Aedes aegypti (Stegomyia aegypti) L. was not detected. The attractants were: (a) a compound found in Aedes eggs (dodecanoic acid); (b) components of nitrogen, phosphorous and potassium‐based (NPK) fertilizer, and (c) infusions of discarded cigarette butts. A solution of ammonium phosphate and potassium nitrate was significantly more attractive to gravid Ae. albopictus than water only. Dodecanoic acid and cigarette butt infusions were not significantly more attractive than the control; however, they attracted various other Diptera and many non‐culicid larvae developed in ovitraps in which these substances were used; thus, the presence of eggs or larvae of other species may have deterred Aedes oviposition. Significantly more Aedes eggs were found in ovitraps under vegetation than in ovitraps placed inside houses or against external walls. Clear‐sided ovitraps in which black mesh was placed over a black ring floating on the water surface collected significantly fewer eggs than black ovitraps with identically placed mesh and rings.     

Evaluation of herbivore-induced plant volatiles for monitoring green lacewings in Washington apple orchards

We evaluated five herbivore-induced plant volatiles plus a male-produced pheromone as attractants for adult green lacewings in Washington apple orchards in 2008. We found at least five attractants or combinations of attractants were attractive to the three most abundant green lacewing species in our trials. Chrysopa nigricornis and Chrysopa oculata were attracted to the combination of methyl salicylate and iridodial with iridodial alone being the second best attractant. Chrysoperla plorabunda was found in lower numbers than C. nigricornis and C. oculata, but did exhibit a significant attraction to benzaldehyde. In mid-summer, we added the herbivore-induced plant volatile squalene to the study and found it to be exceedingly attractive, but only to male C. nigricornis. Whether alone or in combination, squalene attracted 4–5-fold more C. nigricornis than any other compound tested. Our data have revealed C. nigricornis to be an abundant orchard predator that can be readily monitored with squalene-baited traps. Despite the obvious promise of HIPVs in biological control programs, we urge caution in their deployment as large-scale attractants, at least until further studies have investigated potential disruption of natural enemy population dynamics.     

Suppression of the blowfly Lucilia sericata using odour-baited triflumuron-impregnated targets

Abstract. Field trials were carried out in 1995 and 1996 on farms in the south-west of England to assess the extent to which odour-baited targets could be used to suppress populations of the ectoparasitic blowfly, Lucilia sericata , in sheep pastures. Targets were constructed from 41 × 41 cm squares of aluminium sheet, covered by white cloth which had been dipped in a mixture of sucrose solution (50% w/v) and the chitin synthesis inhibitor triflumuron (10% suspension concentrate). Each target was baited with ≈ 300 g of liver and sodium sulphide solution (10%). Three matched sheep farms were used in the trials. In 1995, triflumuron-impregnated targets were placed around the periphery of sheep pastures at one of the farms in late June, at approximately one target per hectare. In 1996, triflumuron-impregnated targets were placed around the periphery of sheep pastures of a second of the farms in early May, at approximately five targets per hectare. Each year, five sticky targets, used to monitor the L. sericata populations, were also placed in fields at the experimental and the other two farms, which acted as controls. In 1995, the results provided some, although inconclusive, evidence that the triflumuron-impregnated targets had reduced the numbers of L. sericata relative to the populations on the two control farms. In 1996, however, the density of L. sericata on the experimental farm was reduced to almost zero and remained significantly lower than on two control farms throughout the period during which the triflumuron-impregnated targets remained in the field. The results are discussed in relation to the use of triflumuron-treated targets as a practical means of controlling L. sericata and sheep blowfly strike.     

Choosy egg parasitoids: Specificity of oviposition-induced pine volatiles exploited by an egg parasitoid of pine sawflies

Generalist parasitoids are well‐known to be able to cope with the high genotypic and phenotypic plasticity of plant volatiles by learning odours during their host encounters. In contrast, specialised parasitoids often respond innately to host‐specific cues. Previous studies have shown that females of the specialised egg parasitoid Chrysonotomyia ruforum Krausse (Hymenoptera: Eulophidae) are attracted to volatiles from Pinus sylvestris L. induced by the egg deposition of its host Diprion pini L. (Hymenoptera: Diprionidae), when they have previously experienced pine twigs with host eggs. In this study we investigated by olfactometer bioassays how specifically C. ruforum responded to oviposition‐induced plant volatiles. Furthermore, we studied whether parasitoids show an innate response to oviposition‐induced pine volatiles. Naïve parasitoids were not attracted to oviposition‐induced pine volatiles. The attractiveness of volatiles from pines carrying eggs was shown to be specific for the pine and herbivore species, respectively (species specificity). We also tested whether not only oviposition, but also larval feeding, induces attractive volatiles (developmental stage specificity). The feeding of D. pini larvae did not induce the emission of P. sylvestris volatiles attractive to the egg parasitoid. Our results show that a specialist egg parasitoid does not innately show a positive response to oviposition‐induced plant volatiles, but needs to learn them. Furthermore, the results show that C. ruforum as a specialist does not learn a wide range of volatiles as some generalists do, but instead learns only a very specific oviposition‐induced plant volatile pattern, i.e., a pattern induced by the most preferred host species laying eggs on the most preferred food plant.     

The effect of pH, salt concentration and temperature on the survival and growth of Listeria monocytogenes

Factorially designed experiments have been used to study the growth and survival of Listeria monocytogenes in different combinations of pH and salt concentrations at ambient and chill temperatures. Survival at low pH and high salt concentration was strongly temperature dependent. The minimum pH values that allowed survival after 4 weeks from an initial 10⁴ cells were 4·66 at 30†C, 4·36 at 10†C and 4·19 at 5†C. These limits were salt dependent, low (4–6%) salt concentrations improved and higher concentrations reduced survival at limiting pH values. The lowest pH that allowed a 100-fold increase in cell numbers within 60 d was 4·66 at 30†C but this was increased to 4·83 at 10†C. At 5†C growth occurred at pH 7·0 but not at pH 5·13. Simple predictive models describing the effect of hydrogen-ion and salt concentration on the time for at least a 100-fold increase in numbers at 10†C and 30†C were constructed after analysis of the results for a least squares fit to a quadratic model. The interactions between salt and hydrogen-ion concentration on growth were found to be purely additive.