Short-term responses of king penguins Aptenodytes patagonicus to helicopter disturbance at South Georgia

The short-term behavioural effects of helicopter overflights on breeding king penguins Aptenodytes patagonicus at South Georgia were examined. Seventeen helicopter overflights were made at altitudes between 230 and 1,768 m (750–5,800 ft) above ground level. Noise from the aircraft engines and helicopter blades increased sound levels in the colony from a background level of 65–69 dB(A) to a maximum mean peak level of 80 dB(A) during overflights. Penguin behaviour changed significantly during all overflights at all altitudes compared to the pre- and post-flight periods. Pre-overflight behaviour resumed within 15 min of the aircraft passing overhead and no chicks or eggs were observed to be taken by predators during overflights. Non-incubating birds showed an increased response with reduced overflight altitude, but this was not observed in incubating birds. Variability in overflight noise levels did not affect significantly the behaviour of incubating or non-incubating birds. Penguins exhibited a reduced response to overflights as the study progressed (despite later flights generally being flown at lower altitudes) suggesting some degree of habituation to aircraft. To minimise disturbance to king penguins we recommend a precautionary approach such that overflights are undertaken at the maximum altitude that is operationally practical, or preferably are avoided altogether.     

Off‐road vehicles affect nesting behaviour and reproductive success of American Oystercatchers Haematopus palliatus

As human populations and associated development increase, interactions between humans and wildlife are occurring with greater frequency. The effects of these interactions, particularly on species whose populations are declining, are of great interest to ecologists, conservationists, land managers and natural resource policy‐makers. The American Oystercatcher Haematopus palliatus, a species of conservation concern in the USA, nests on coastal beaches subject to various forms of anthropogenic disturbance, including aircraft overflights, off‐road vehicles and pedestrians. This study assessed the effects of these human disturbances on the incubation behaviour and reproductive success of nesting American Oystercatchers at Cape Lookout National Seashore, on the Atlantic coast of the USA. We expanded on‐going monitoring of Oystercatchers at Cape Lookout National Seashore by supplementing periodic visual observations with continuous 24‐h video and audio recording at nests. Aircraft overflights were not associated with changes in Oystercatcher incubation behaviour, and we found no evidence that aircraft overflights influenced Oystercatcher reproductive success. However, Oystercatchers were on their nests significantly less often during off‐road vehicle and pedestrian events than they were during control periods before the events, and an increase in the number of off‐road vehicles passing a nest during incubation was consistently associated with significant reductions in daily nest survival (6% decrease in daily nest survival for a one‐vehicle increase in the average number of vehicles passing a nest each day; odds ratio = 0.94; 95% confidence interval (CI) 0.90, 0.98) and hatching success (12% decrease in hatching success for a one‐vehicle increase in the average number of vehicles passing a nest each day; odds ratio = 0.88; 95% CI 0.76, 0.97). Management of vehicles and pedestrians in areas of Oystercatcher breeding is important for the conservation of American Oystercatchers.     

Comparative nesting and feeding ecology of skylarks Alauda arvensis on arable farmland in southern England with special reference to set-aside

1. A study of skylark Alauda arvensis L. breeding ecology in relation to crop type was carried out from April to August 1992 on arable land in southern England. Set-aside land was included in this comparative study.
2. Territory density averaged 0·15 ha⁻¹. It was 2–3 times higher in fields of set-aside and grass, especially permanent pasture, than in winter and spring-sown cereals.
3. Territory size was nearly twice as large in fields of winter cereals (4·5 ha) than in other crop types (2·5 ha). Where set-aside was present on one farm, territory size in set-aside (1·7 ha) was a third lower than in cereals and grass.
4. Nesting began in set-aside and permanent pasture in April and peaked in late May. Nesting was not detected in spring barley until late May and in silage grass until early June. The density of successful nests in set-aside fields was more than double that in any of the arable crop types.
5. Average clutch size at hatching was 3·91 eggs in fields of set-aside, over 15% higher than in silage grass (3·40) and in spring barley (3·27).
6. Fledging success did not differ according to crop type, but productivity, expressed as the number of fledglings produced per hectare, was 0·50 in set-aside, 0·13 in silage grass, and 0·21 in spring barley. Nests with chicks were not found in fields of winter cereals. The causes of chick death were thought to be predation in set-aside fields, farming practices in silage grass fields, and suspected starvation in spring cereals.
7. The potentially high nesting success of skylarks in set-aside implies that sympathetic set-aside management could play an important part in reversing its decline across the European Union.     

Effects of predator exposure on baseline and stress-induced glucocorticoid hormone concentrations in pumpkinseed Lepomis gibbosus

We compared baseline and maximal cortisol concentrations between predator exposure and prey blood samples in pumpkinseed Lepomis gibbosus, captured using a standardised fishing event underneath osprey Pandion haliaetus nests and away from osprey nests. We did not detect differences in cortisol or glucose between sites. These findings suggest that predictable sources of predation risk may not confer stress-related costs in teleosts.     

Influence of pregnancy on the thermal biology of the lizard, Sceloporus jarrovi: why do pregnant females exhibit low body temperatures?

1. Selected body temperatures of female lizards, Sceloporus jarrovi , were measured on a photothermal gradient during late pregnancy and again when postpartum, and pregnant females were subjected to one of three fluctuating temperature regimes that simulated body temperatures of (1) pregnant females, (2) postpartum females or (3) allowed normal thermoregulation.
2. Overall, females selected lower body temperatures when pregnant (mean = 32·0°C) than when postpartum (mean = 33·5°C).
3. Females regulated body temperature more precisely when pregnant than when postpartum as judged by their smaller variances in body temperature throughout the day.
4. When pregnant, females selected a lower mean maximum body temperature (mean: pregnant = 32·8°C; postpartum = 34·5°C) than when postpartum, but selected mean minimum body temperatures did not differ.
5. None of the experimental temperature treatments was detrimental to pregnant females. Female body length increased during pregnancy but the rate of increase did not differ among treatments. Moreover, length-adjusted body mass of postpartum females did not differ among treatments.
6. Pregnant females that experienced postpartum body temperatures produced neonates that were smaller in body mass and length than pregnant females that experienced pregnant body temperatures and females that were allowed to thermoregulate.
7. For neonates resulting from the postpartum body temperature treatment, the disparity in the body length, but not mass, was still observed at 9 days of age, although survival and growth of neonates was high and did not differ among treatments.
8. The results demonstrate that pregnant females could maintain higher postpartum body temperatures without compromising their physical condition, but select relatively low body temperatures, presumably to avoid decrements in offspring fitness.     

A method for tracking the behaviour of mature and immature salmon parr around nests during spawning

A remote monitoring system was developed to provide information on the behaviour of mature and immature Atlantic salmon Salmo salar parr at nests during the spawning season. An octagonal passive integrated transponder (PIT) detector (0·865 m maximum diameter) designed to surround nests of Atlantic salmon was used to identify individual salmon parr present at 38 spawning events in three circular spawning channels. The range of the detector for PIT tags presented in the optimum orientation was 2·4 cm (range between tags 1·7–3·0 cm). Using a sub-sample of 20 spawnings, the mean efficiency of the detector (number of fish passes registered relative to number of passes observed on video) was 70·5% (range 32-100%). There were no significant effects of time from spawning, total number of registrations, body size or maturity status (mature or immature) on efficiency. However, fish were more likely to be detected entering nests than leaving, as departures were more rapid and higher in the water column. The PIT detector did not affect the numbers of parr at spawnings or between spawning intervals of females, and allowed for the individual identification of 65 of the 72 parr observed in nests during spawning. In all cases where certain identifications were not possible and the video was of satisfactory quality, this was due to obstruction of the camera view by anadromous fish. The remote monitoring system was thus effective in identifying behavioural differences, and only one of 20 immature parr was ever detected during the period encompassing 10 min before and after spawning compared with 30 or 40 mature parr.     

Density,nest site characteristics and breeding rates of the osprey (Pandion haliaetus) in the southern limit of its range in the Western Palearctic (Boa Vista,Cape Verde Islands)

Between 2004 and 2007, we studied density, habitat features and breeding parameters of the osprey (Pandion haliaetus) population in Boa Vista Island (Cape Verde). A total of 79 nest structures were identified, 37 of which were occupied for at least 1 year during the study period. The osprey population ranged between 14 and 18 pairs, and the mean density and distance between neighbouring occupied nests were 2.58 pairs per 100 km² and 3089 m, respectively. Occupied nests were found to be significantly further from the coastline and roads than unoccupied nests, but the distances from villages were similar. The majority (81.1%) of the 37 occupied nests were easily accessible to humans. Mean clutch size was 2.59, average productivity was 0.76 young/active nest, and breeding success was 58.8% [Correction added on 13 May 2013, after first online publication: the average productivity was changed from 0.72 to 0.76]. Density in Boa Vista was higher than that in other sedentary island populations in the Western Palearctic, whereas the productivity was the lowest of this region. Clutch size did not vary among Western Palearctic populations, but the differences observed in productivity were likely influenced by local factors that in Boa Vista are attributed to nest depredation by the brown‐necked raven (Corvus ruficollis) and to direct human persecution.     

Improving grasslands: the influence of soil moisture and nitrogen fertilization on the establishment of seedlings

1. In order to investigate the factors influencing the establishment of seedlings in permanent grassland, the influence of soil moisture and nitrogen fertilization on competition between established plants of Lolium perenne and seedlings of Phleum pratense or Trifolium pratense was studied in two experiments under greenhouse conditions using the 'split-box'-technique.
2. There was no difference in the production of plant dry matter of P. pratense or T. pratense between 30% volumetric soil water content (−0·005 MPa) and 22% (−0·04 MPa), but 15% soil moisture (−0·33 MPa) reduced plant growth. L. perenne yields were linearly reduced by reduced soil moisture content.
3. Shoot competition from L. perenne reduced the plant dry matter yield of P. pratense and T. pratense more than did root competition in these experiments. When shoot competition was present, differences between moisture contents were not detected, indicating that light was probably the limiting resource under such conditions. No significant interaction between root competition and soil moisture was observed for plant weight.
4. Root competition was not prevented even though sufficient water and nitrogen were supplied. This indicated either that some other growth factor was limiting or the plants competed for resources at the root hair level even though sufficient resources were supplied at the pot or field scale. Therefore, in the situation of direct drilling of species during grassland renovation, it may be difficult to alleviate competition by adequate provision of water and nitrogen.     

Hosts feeding only Brown‐headed Cowbird fledglings: where are the host fledglings?

ABSTRACT.   Brown-headed Cowbirds ( Molothrus ater ) impose costs on their hosts, usually resulting in fewer or no host young being raised. In 95% of the studies where attempts were made to estimate the cost of parasitism, cost was measured as the difference in host nestling survival in parasitized and unparasitized nests. For hosts that fledge at least some of their own young along with cowbirds, the magnitude of further brood reduction beyond the nestling stage is not known. To examine the possibility that further brood reduction occurs during the fledgling stage, we compiled 102 observations of 45 species feeding fledgling cowbirds. In 97 cases, observers did not record hosts feeding their own young, suggesting that many young die shortly after fledging from parasitized nests. These observations suggest that measuring the cost of parasitism as the difference between the number of host young fledged from parasitized and unparasitized nests probably overestimates the reproductive output of many hosts. If costs increase during the fledgling stage, then the costs of brood parasitism have been underestimated in many studies.     

Spatial patterns, temporal variability, and the role of multi-nest colonies in a monogynous Spanish desert ant

Abstract 1. The colonies of the Spanish desert ant Cataglyphis iberica are polydomous. This study describes the temporal and spatial patterns of the polydomy in this species at two different sites, and presents analyses of its role in reducing the attacks of the queen over sexual brood, and in allowing better habitat exploitation.
2. The spatial distribution of nests was clumped while colonies were distributed randomly. Mean nearest neighbour distance ranged from 3.4 to 7.0 m for nests and from 12.3 to 14.1 m for colonies. Distance of foragers searching for food varied among nests: mean values were between 6.1 and 12.6 m.
3. At both sites, the maximum number of nests per colony occurred in summer, during the maximum activity period of the species. Colonies regrouped at the end of this period but overwintered in several nests.
4. Nest renewal in C. iberica colonies was high and showed great temporal variability: nests changed (open, close, re-open) continuously through the activity season and/or among years. The lifetime of up to 55% of nests was only 1–3 months.
5. Polydomy in C. iberica might decrease the interactions between the queen and the sexual brood. In all colonies excavated just before the mating period, the nest containing the queen did not contain any virgin female. Females were in the queenless nests of the colony.
6. The results also suggest that polydomous C. iberica colonies may enhance habitat exploitation because foraging activity per colony increases with nest number. The relationship between total prey input and foraging efficiency and number of nests per colony attains a plateau or even decreases after a certain colony size (four to six nests). This value agrees with the observed mean number of nests per colony in C. iberica .     

Mercury Concentrations in Tissues of Osprey From the Carolinas,USA

Abstract: Mercury (Hg) contamination is believed to be one of the most significant pollution hazards to wildlife in the southeastern United States, yet comprehensive studies of Hg contamination of piscivorous raptors are rare in this region. We analyzed total Hg (THg) concentration in tissues of 39 osprey (Pandion haliaetus) primarily from coastal counties of South Carolina, USA, to describe tissue distribution of Hg and to determine whether age or sex influenced Hg accumulation. To determine whether Hg poses health risks to osprey breeding in this region, we also measured selenium in all tissues and the percentage of THg that was methylated in a subset of individuals. Osprey with adult plumage tended to have higher and more variable Hg concentrations in their tissues than younger birds. Whereas highest concentrations of Hg were found in liver and kidney of older birds, chicks had highest concentrations in keratinized tissues. Mercury concentrations were correlated between feathers and soft tissues, but talon concentrations of Hg were better correlated with organs than to feathers in most cases. Contrary to previous studies on birds, we found no relationship between Hg concentration in primary feathers and the sequence in which the feather was molted. We attribute this observation to the irregular and protracted molting pattern of osprey. Also contrary to other studies, feather concentrations of Hg were considerably lower than concentrations in liver and kidney. Osprey with high concentrations of Hg in their livers and kidneys accumulated as much as 99% of it as Hg(II), suggesting that demethylation and sequestration of Hg(II) may be even more critical to mitigating adverse effects than it is for other birds that eliminate most of their Hg burden in feathers. In addition, selenium was co-sequestered with Hg in the liver and kidneys and may further mitigate any adverse effects. Based on these findings, we suggest that most osprey in this region are not currently at risk of Hg toxicosis, but recommend that additional ecotoxicological studies be performed to monitor risk to osprey in this coastal region facing heavy development. We also suggest that concentrations of Hg in talon and claw may serve as important indicators of previous exposure and provide useful information for natural resource managers seeking to assess health risks to birds.     

The functional relationship between peak spring floods and survival and growth of juvenile Atlantic Salmon (Salmo salar) and Brown Trout (Salmo trutta)

1. The effects of high spring floods on survival and growth of juvenile Atlantic Salmon, Salmo salar, and Brown Trout, Salmo trutta , are explored, using data from a long-term study in the River Saltdalselv, northern Norway. The flow regime in this river is typical for northern rivers.
2. There was considerable variation in year class strength of both species.
3. Mortality of Atlantic Salmon increased significantly in years with high discharge during the alevin stage as well as the first week after emergence. High discharge during the egg stage and more than 1 week after emergence seemed to be of minor importance. Water temperature at emergence was rather high (average 10·5 °C) and did not significantly affect year class strength.
4. Brown Trout emerged earlier than Atlantic Salmon at an average water temperature of 8·2 °C. Highest mortality was observed in years with low water temperatures at emergence as well as high discharge during the alevin stage.
5. For 1-year-old fish or older, the size of the spring peak flood did not influence mortality significantly.
6. Growth of Atlantic Salmon parr was diminished in years with a high peak spring flood. A similar effect on Brown Trout was not detected.     

AIRCRAFT SOUND AND DISTURBANCE TO BOWHEAD AND BELUGA WHALES DURING SPRING MIGRATION IN THE ALASKAN BEAUFORT SEA

Short‐term behavioral responses of bowhead whales (Balaena mysticetus) and beluga whales (Delphinapterus leucas) to a Bell 212 helicopter and Twin Otter fixed‐wing aircraft were observed opportunistically during four spring seasons (1989–1991 and 1994). Behaviors classified as reactions consisted of short surfacings, immediate dives or turns, changes in behavior state, vigorous swimming, and breaching. The helicopter elicited fewer detectable responses by bowheads (14% of 63 groups) than by belugas (38% of 40). Most observed reactions by bowheads (63%) and belugas (86%) occurred when the helicopter was at altitudes ≤150 m and lateral distances ≤250 m. Belugas reacted significantly more frequently during overflights at lateral distances ≤250 m than at longer lateral distances (P= 0.004). When the helicopter was on the ice with engines running, 7 of 14 groups of belugas reacted, up to 320 m away, sometimes with small‐scale (≤100 m) diversion; only 1 of 8 groups of bowheads reacted. For the fixed‐wing aircraft, few bowheads (2.2%) or belugas (3.2%) were observed to react to overflights at altitudes 60–460 m. Most observed reactions by bowheads (73%) and belugas (70%) occurred when the fixed‐wing aircraft was at altitudes ≤182 m and lateral distances ≤250 m. However, the proportions reacting, especially to low‐altitude flights (e. g., ≤182 m), were underestimated for both species because observation opportunities were brief. Even so, reactions were more common when the aircraft was low (≤182 m): P= 0.009 for belugas, P= 0.06 for bowheads. There was little if any reaction by bowheads when the aircraft circled at altitude 460 m and radius 1 km. Aircraft sounds measured underwater at depths 3 m and 18 m showed that a Bell 212 helicopter was 7–17.5 dB noisier than a Twin Otter (10–500 Hz band). Bell 212 sound consisted mainly of main rotor tones ahead of the helicopter and tail rotor tones behind it. Twin Otter sound contained fewer prominent tones. Peak sound level as received underwater was inversely related to aircraft altitude, and received levels at 3 m depth averaged 2.5 dB higher than at 18 m depth. The dominant low‐frequency components of aircraft sound are presumed to be readily audible to bowheads. For belugas, these components may be inaudible, or at most only weakly audible. Mid‐frequency sound components, visual cues, or both, are probably important in eliciting beluga reactions to aircraft.     

Growth and reducing capacity of Listeria monocytogenes under different initial redox potential

Aims:  To determine the reducing capacity of Listeria monocytogenes and to highlight the effect of redox potential on its growth parameters.
Methods and Results:  The reducing capacity of L. monocytogenes was monitored in Brain Heart Infusion Broth media at different initial redox potential (Eh) and pH at 37°C. The effect of Eh obtained by gas flushing (air, N₂ and N₂-H₂) or by adding potassium ferricyanide and dithiotreitol in concentration from 1 to 10 mmol l⁻¹on L. monocytogenes growth parameters at pH 6·0, 7·0 and 8·0 was investigated. A total change of 539 mV (±44 mV) from an initial redox value of +330 ± 8 mV to a more negative potential in redox curves was observed resulting from L. monocytogenes growth at pH 7·0 at 37°C. A significant influence of pH and redox potential on L. monocytogenes lag phase of growth was shown ( P  < 0·05).
Conclusions:  Listeria monocytogenes exhibited longer lag phase in reducing conditions and at pH 6·0. The method used to modify the redox potential was shown to have no effect on growth parameters at pH 7·0.
Significance and Impact of the Study:  The provided information on the extending lag time and the possible delayed growth of this major pathogen in reducing conditions might be useful for its control in foods.     

Resources exploitation by ants facilitates lizard egg survival

Abstract.  1.  This study investigated the interaction between eggs of the lizard Mabuya longicaudata Hallowell and two species of ant: a lizard egg commensalist ( Paratrechina longicornis Latreille), and an egg predator ( Pheidole taivanensis Forel). The ecological interaction between P. longicornis and lizard eggs was tested, and it was predicted that when the interaction was removed, lizard eggs would be attacked by P. taivanensis .
2. Field observations showed that both ant species actively searched for lizard eggs, and that P. taivanensis typically found new lizard nests earlier than P. longicornis did. Left undisturbed, P. taivanensis predation dramatically reduced lizard egg survival. While P. longicornis usually found nests later, they were able to displace P. taivanensis . As a result, proportional egg survival was higher in nests with P. longicornis (0.95 ± 0.04 eggs) compared to nests without either ant species (0.65 ± 0.09 eggs), or nests occupied by P. taivanensis (0.07 ± 0.02 eggs).
3. When P. longicornis ants were experimentally excluded from lizard nests, the proportion of eggs surviving significantly decreased because of increased P. taivanensis predation. Paratrechina longicornis benefits from water that condenses on the eggs. When this resource disappears P. longicornis abandons the nest, leading to predation of the eggs by P. taivanensis .     

Influence of body condition and forage type on prevalence of Escherichia coli O157:H7 and Salmonella in grazing beef cows

Aim:  To determine the influence of body condition (BC) and forage type on the prevalence of faecal shedding of Escherichia coli O157:H7 and Salmonella from beef cows.
Methods and Results:  Thin or moderately conditioned cows ( n  =   115) were randomly assigned to graze either common bermudagrass ( n  =   3 pastures) or toxic endophyte-infected tall fescue ( n  =   3 pastures) for 62 days. Faecal samples were collected on day 0, 30 and 62. Overall percentage of faecal samples positive for E. coli O157:H7 was 2·6% and 2·0% for Salmonella . Percentage of cows positive for both E. coli O157:H7 and Salmonella on at least one occasion was 6·1%. BC, forage type or the interaction did not influence the prevalence of E. coli O157:H7 or Salmonella in the faeces of cows.
Conclusions:  BC at initiation of the grazing period or loss of BC in moderate conditioned cows during the grazing period did not influence faecal shedding of E. coli O157:H7 or Salmonella . Consumption of either forage type did not influence faecal shedding of either E. coli O157:H7 or Salmonella in beef cows of thin or moderate BC.
Significance and Impact of the Study:  Change in BC that typically occurs during the normal production cycle in grazing cows did not influence faecal shedding of pathogenic bacteria regardless of forage type.     

Effects of experimental small-scale grassland fragmentation on spatial distribution, density, and persistence of ant nests

Abstract.  1. Grassland fragmentation is expected to influence the abundance of different invertebrate species to a different extent. Fragmentation-related effects are of particular importance in species that interact with many other species.
2. The density and spatial distribution of nests of 15 ant species in experimentally fragmented calcareous grasslands at three sites in the Northern Swiss Jura mountains were examined. Fragments of different size (0.25 m², 2.25 m², and 20.25 m²) were isolated by a 5-m wide strip of frequently mown vegetation. Control plots of corresponding size were situated in adjacent undisturbed grassland.
3. Three years after initiation of the experiment, ant nest density did not differ between fragments and control plots. Six years after initiation of the experiment, however, ant nest density and forager abundance were higher in large fragments than in large control plots. Ant nests tended to occur more frequently along the edge of fragments than in the core area. Persistence time of nests of the most abundant species, Lasius paralienus , tended to be shorter in fragments than in control plots. Furthermore, persistence time was longer in nests situated close to the fragment edge than in nests in the core area.
4. Effects on nest density, edge effects on the spatial distribution of nests, and the relationships between nest density and environmental factors were more pronounced when only nests of L. paralienus were considered. The implications of these findings for plant and other invertebrate species are discussed.     

The risk of Listeria monocytogenes infection in beef cattle operations

Aim:  To determine the prevalence of Listeria monocytogenes and associated risk factors among beef operations (cow-calf and feedlot) in central and southern California.
Methods and Results:  A repeated cross-sectional study where faecal and environmental samples were collected from 50 operations three times a year at different seasons was carried out. Samples were tested for presence of L. monocytogenes using a combination of enrichment and polymerase chain reaction tests. Data on putative risk factors were also collected. Listeria monocytogenes was detected in faecal samples from cows, calves and other animals on calf-cow operations at proportions of 3·1%, 3·75% and 2·5%, respectively. The organism was detected in 5·3% of cut-grass, 5·3% of soil, 14·3% of irrigation ditches, 3·1% of the ponds and 6·5% of water troughs samples. Listeria monocytogenes was less common in faecal (0·3%) and soil (0·75%) samples collected from feedlots.
Conclusions:  Listeria monocytogenes was present at a higher proportion among cow-calf operations than feedlots. There was no significant seasonal variation in the occurrence of this pathogen within the two types of operations.
Significance and Impact of the Study:  If risk mitigation strategies were implemented to reduce the public health risk these should focus in cow-calf operations.     

Survival and growth of Cronobacter species (Enterobacter sakazakii) in wheat-based infant follow-on formulas

Aims:  To determine the survival and growth characteristics of Cronobacter species ( Enterobacter sakazakii ) in infant wheat-based formulas reconstituted with water, milk, grape juice or apple juice during storage.
Methods and Results:  Infant wheat-based formulas were reconstituted with water, ultra high temperature milk, pasteurized grape or apple juices. The reconstituted formulas were inoculated with Cronobacter sakazakii and Cronobacter muytjensii and stored at 4, 25 or 37°C for up to 24 h. At 25 and 37°C, Cronobacter grew more (>5 log₁₀) in formulas reconstituted with water or milk than those prepared with grape or apple juices ( c. 2–3 log₁₀). The organism persisted, but did not grow in any formulas stored at 4°C. Formulas reconstituted with water and milk decreased from pH 6·0 to 4·8–5·0 after 24 h, whereas the pH of the formulas reconstituted with fruit juices remained at their initial pH values, c. pH 4·8–5·0.
Conclusions:  Cronobacter sakazakii and C. muytjensii can grow in reconstituted wheat-based formulas. If not immediately consumed, these formulas should be stored at refrigeration temperatures to reduce the risk of infant infection.
Significance and Impact of the Study:  The results of this study will be of use to regulatory agencies and infant formula producers to recommend storage conditions that reduce the growth of Cronobacter in infant wheat-based formulas.     

Evaluation of rapid methods for the determination of okadaic acid in mussels

L.CROCI, A.STACCHINI, L.COZZI, G.CICCAGLIONI, F.MAZZEI, F.BOTRÈ AND L.TOTI. 2001 .
Aims: Two different screening methods, a Buffalo Green Monkey cytotoxicity test and a biosensor test, have been considered to replace the official mouse bioassay in monitoring for okadaic acid (OA) levels in mussels.
Methods and Results: Diarrhoetic shellfish poison-contaminated mussels from the Adriatic Sea were assayed in parallel by means of the mouse bioassay and both alternative methods. Both the cytotoxicity test and the biosensor test showed high sensitivity (OA 0·01 mg g^–1 hepatopancreas and 0·002 mg g^–1 hepatopancreas, respectively) and a high correlation with the mouse bioassay ( r =0·932, P  < 0·001 and r =− 0·850, P  < 0·001, respectively).
Conclusions: Both methods are efficacious, quick, inexpensive and provide data on the amount of toxin present in mussels.
Significance and Impact of the Study: Both methods, besides allowing the simultaneous assay of a great number of samples, comply with the ethical need to reduce the use of animals in the laboratory.