Abstract Epilepsy, a clinical diagnosis characterized by paroxysmal episodes known as seizures, affects 1% of people worldwide. Safe and patient-specific treatment is vital and can be achieved by the development of rapid pre-clinical models of for identified epilepsy genes. Epilepsy can result from either brain injury or gene mutations, and can also be induced chemically. Xenopus laevis tadpoles could be a useful model for confirmation of variants of unknown significance found in epilepsy patients, and for drug re-purposing screens that could eventually lead to benefits for patients. Here, we characterise and quantify seizure-related behaviours in X. laevis tadpoles arrayed in 24-well plates. To provoke acute seizure behaviours, tadpoles were chemically induced with either pentylenetetrazole (PTZ) or 4-aminopyridine (4-AP). To test the capacity to adapt this method for drug testing, we also exposed induced tadpoles to the anti-seizure drug valproate (VPA). Four induced seizure-like behaviours were described and manually quantified, and two of these (darting, circling) could be accurately detected automatically, using the video analysis software TopScan. Additionally, we recorded swimming trajectories and mean swimming velocity. Automatic detection showed that either PTZ or 4-AP induced darting behaviour and increased mean swimming velocity compared to untreated controls. Both parameters were significantly reduced in the presence of VPA. In particular, darting behaviour was a shown to be a sensitive measure of epileptic seizure activity. While we could not automatically detect the full range of seizure behaviours, this method shows promise for future studies, since X. laevis is a well-characterised and genetically tractable model organism.
Summary Potassium tellurite was assessed for the selection of verocytotoxigenic (VT+) Escherichia coli O157. MICs were higher for VT+ E. coli O157 than for other strains of E. coli and for Aeromonas spp. MacConkey medium containing sorbitol, tellurite and cefixime (TC-SMAC) permitted the growth of VT+ E. coli O157 and Shigella sonnei but partially or completely inhibited the growth of 67% of other strains of E. coli and all or most strains of other sorbitol-non-fermenting species tested. Of 391 rectal swabs from cattle screened on TC-SMAC medium, 26 yielded isolates of VT+ E. coli O157 whereas sorbitol-MacConkey medium with cefixime and rhamnose yielded only nine isolates. Inclusion of potassium tellurite in sorbitol-MacConkey agar markedly increased the rate of isolation of VT+ E. coli O157 from cattle rectal swabs and may do so for other types of specimen.
Summary: Increasing numbers of cases of Vero cytotoxin producing Escherichia coli (VTEC) O157 infection, well publicised incidents, and new scientific evidence make it appropriate to produce new guidelines for their control. This document reviews the clinical and epidemiological features of VTEC O157 infection, describes the principles of microbiological investigation and laboratory safety, and presents recommendations for the prevention of spread of VTEC) O157. The recommendations consider direct spread of infection from animals, foodborne spread, the institutions in which spread is more likely to occur (nursing homes, schools, and children’s day nurseries), and groups at particular risk of acquiring and transmitting infection (in essence, food handlers, and those unable to maintain high standards of hygiene for themselves and their carers).
Summary Cattle arriving for slaughter at abattoirs in the Veneto region of N. Italy were examined for intestinal carriage of Escherichia coli O157. Rectal swabs were cultured in modified buffered peptone water and E. coli O157 was concentrated by an immunomagnetic separation technique; the magnetic beads were cultured onto cefixime tellurite sorbitol MacConkey agar. Sorbitol non‐fermenting E. coli O157 was isolated from 15 (3.6%) of 419 feedlot cattle but not from 437 veal calves or 65 culled cows. All strains of E. coli O157 hybridized with DNA probes specific for the VT 1 or VT 2 genes, but two strains did not produce toxin detectable by Vero cell assay. Six different plasmid profiles were observed with all strains harbouring the large 93 kb plasmid characteristic of VTEC. Six strains produced urease but otherwise strains were biochemically typical of E. coli O157. One strain was resistant to streptomycin, tetracycline and sulphonamides but the remainder were sensitive to all antimicrobials tested. This is the first description of the isolation of verocytotoxin‐producing E. coli O157 from cattle in Italy. As the contamination of bovine carcasses with E. coli O157 during slaughter and processing has been demonstrated, the risk of transmission of this organism from beef cattle to the human population in the Veneto region, through foods of bovine origin or by other routes, should not be overlooked.
Protozoa morphologically consistent with Caryospora sp. are one of the few pathogens associated with episodic mass mortality events involving free-ranging sea turtles. Parasitism of green turtles (Chelonia mydas) by these coccidia and associated mortality was first reported in maricultured turtles in the Caribbean during the 1970s. Years later, epizootics affecting wild green turtles in Australia occurred in 1991 and 2014. The first clinical cases of Caryospora-like infections reported elsewhere in free-ranging turtles were from the southeastern US in 2012. Following these initial individual cases in this region, we documented an epizootic and mass mortality of green turtles along the Atlantic coast of southern Florida from November 2014 through April 2015 and continued to detect additional, sporadic cases in the southeastern US in subsequent years. No cases of coccidial disease were recorded in the southeastern US prior to 2012 despite clinical evaluation and necropsy of stranded sea turtles in this region since the 1980s, suggesting that the frequency of clinical coccidiosis has increased here. Moreover, we also recorded the first stranding associated with infection by a Caryospora-like organism in Hawai’i in 2018. To further characterize the coccidia, we sequenced part of the 18S ribosomal and mitochondrial cytochrome oxidase I genes of coccidia collected from 62 green turtles found in the southeastern US and from 1 green turtle found in Hawai’i. We also sequenced the ribosomal internal transcribed spacer regions from selected cases and compared all results with those obtained from Caryospora-like coccidia collected from green turtles found in Australia. Eight distinct genotypes were represented in green turtles from the southeastern US. One genotype predominated and was identical to that of coccidia collected from the green turtle found in Hawai’i. We also found a coccidian genotype in green turtles from Florida and Australia with identical 18S and mitochondrial sequences, and only slight inter-regional differences in the internal transcribed spacer 2. We found no evidence of geographical structuring based on phylogenetic analysis. Low genetic variability among the coccidia found in green turtle populations with minimal natural connectivity suggests recent interoceanic dissemination of these parasites, which could pose a risk to sea turtle populations.
A case-controlled study of the incidence of heat-labile enterotoxin-producing Escherichia coli (LT+ETEC) and other enteric pathogens in travellers returning to the Sheffield area was conducted from May 1984 to April 1986. LT+ETEC were found in 35 (5.8%) of 600 travellers to developed countries (mainly popular Mediterranean holiday resorts), 36 (11.3%) of 320 travellers to less-developed countries, and 11 (0.9%) of 1282 control patients whose illness was not associated with recent travel abroad. A seasonal peak of LT+ETEC infection was observed only in travellers to developed countries, with infections being significantly commoner in August to October. There was no significant deviation from expected age/sex distribution of LT+ETEC infection. Strains of LT+ETEC from travellers produced more toxin than strains from control patients, strains from travellers to less-developed countries producing most of all.
Abstract – A population of the exotic pest fish Gambusia holbrooki inhabiting a drainage channel was sampled regularly to record responses to flooding and subsequent population reestablishment. The flood reduced numbers in the channel to near‐zero levels. After remaining very low for 2 months, densities increased steadily through juvenile recruitment and the concentration of fish in drying pools. Tagging revealed that in general, movements between pools separated by only a few metres were relatively limited. There was evidence for habitat segregation and population subdivision, as fish from different pools varied markedly in terms of mean population density, movement behaviour, sex and size composition and juvenile recruitment. The most favoured pool was relatively deep, well lit and had the greatest habitat diversity. Gambusia control measures are likely to be most effective if timed to coincide with floods (which reduce local population densities) and/or droughts (which concentrate fish and allow targeting of source populations).
Abstract Epilepsy, a clinical diagnosis characterised by paroxysmal episodes known as seizures, affects 1% of people worldwide. Safe and patient‐specific treatment is vital and can be achieved by the development of rapid pre‐clinical models of for identified epilepsy genes. Epilepsy can result from either brain injury or gene mutations, and can also be induced chemically. Xenopus laevis tadpoles could be a useful model for confirmation of variants of unknown significance found in epilepsy patients, and for drug re‐purposing screens that could eventually lead to benefits for patients. Here, we characterise and quantify seizure‐related behaviours in X. laevis tadpoles arrayed in 24‐well plates. To provoke acute seizure behaviours, tadpoles were chemically induced with either pentylenetetrazole (PTZ) or 4‐aminopyridine (4‐AP). To test the capacity to adapt this method for drug testing, we also exposed induced tadpoles to the anti‐seizure drug valproate (VPA). Four induced seizure‐like behaviours were described and manually quantified, and two of these (darting, circling) could be accurately detected automatically, using the video analysis software TopScan. Additionally, we recorded swimming trajectories and mean swimming velocity. Automatic detection showed that either PTZ or 4‐AP induced darting behaviour and increased mean swimming velocity compared to untreated controls. Both parameters were significantly reduced in the presence of VPA. In particular, darting behaviour was a shown to be a sensitive measure of epileptic seizure activity. While we could not automatically detect the full range of seizure behaviours, this method shows promise for future studies since X. laevis is a well‐characterised and genetically tractable model organism. image
Summary In May-June 1992 cases of infection with verocytotoxin-producing (VT+) Escherichia coli O157 in South Yorkshire could have been associated with prior consumption of beef from a local abattoir. During investigation of the abattoir, bovine rectal swabs and samples of meat and surface swabs from beef carcasses were examined for E. coli O157, isolates of which were tested for toxigenicity, plasmid content and phage type. E. coli O157 was isolated from 84 (4%) of 2103 bovine rectal swabs; of these 84, 78 (93%) were VT+, the most common phage types being 2 and 8, the types implicated in the cluster of human cases. Positive cattle were from diverse sources within England. E. coli O157 was isolated from 7 (30%) of 23 carcasses of rectal swab-positive cattle and from 2 (8%) of 25 carcasses of rectal swab-negative cattle. The study has shown that cattle may be a reservoir of VT+ E. coli O157, and that contamination of carcasses during slaughter and processing may be how beef and beef products become contaminated and thereby transmit the organism to man.
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