Phenotypic and genetic characterization of enteropathogenic Escherichia coli (EPEC) and entero-aggregative E. coli (EAEC) from diarrhoeal and non-diarrhoeal children in Libya

A total of 50 Escherichia coli strains isolated in a Libyan hospital (20 from children with diarrhoea and 30 from healthy children) were investigated for their pathotypes and virulence traits. Altogether nine eae-positive (enteropathogenic E. coli, EPEC) and nine aggR-positive (entero-aggregative E. coli, EAEC) strains were identified. Significantly (P=0.001) more EPEC strains were identified from diarrhoeal patients (n=8) than from healthy controls (n=1), while six EAEC strains were identified from diarrhoeal and three from healthy children. Typical (eae(+), EAF(+), bfp(+)) EPEC strains (n=6) belonged to classical EPEC serogroups O55, O114, O127 and showed localized adherence on Hela cells. EAEC strains revealed genetic heterogeneity but uniformly adhered to HeLa cultures in an entero-aggregative adherence pattern. Antibiotic resistance frequently, characterized the strains. Sixty-eight percentage of the strains were resistant against at least one antibiotic and 30% harbored a class 1 integron independently of their clinical background. This is the first report from North Africa demonstrating the significance of EPEC and EAEC.     

Chicken dendritic cells and type II pneumocytes express a common intracellular epitope

1. CVI-ChNL 74·3, a dendritic cell-specific monoclonal antibody (mAb) also identifies chicken lung granular pneumocytes (type II pneumocytes), which produce surfactant.

2. The 74·3 mAb does not cross-react with any other avian or mammalian granular pneumocyte, and provides a convenient tool for monitoring the status of type II pneumocytes in the chicken lung.     

Colonisation of conventional weaned pigs by enteropathogenic Escherichia coli (EPEC) and its hazard potential for human health

Enteropathogenic Escherichia coli (EPEC) bacteria frequently cause severe enteric diseases primarily in children and in young rabbits. Their pathogenicity for pigs has been tested by oral infection of colostrum-deprived newborn, and of severely immunosuppressed weaned pigs, but colonisation of conventional weaned pigs by porcine EPEC has not been experimentally studied. EPEC show similarities to enterohaemorrhagic E. coli (EHEC) additionally carrying shiga toxin genes integrated into the chromosome by lambdoid phages. We have demonstrated earlier that the porcine EPEC prototype strain P86-1390 (O45) could be transduced in vivo (in ligated loops of weaned pigs), by Stx2 phage derived from a human EHEC. Thus, the ability of this porcine EPEC strain to colonise conventional weaned pigs under farming conditions became a question of relevance to human health. To clarify this question, four intragastric infection experiments were performed on a total of 95 conventional weaned pigs. The EPEC P86-1390 and other well-characterised porcine EPEC strains were applied to 54 pigs, leaving 41 weaned pigs as negative controls. In three experiments moderate predispositions were applied: coinfections with enterotoxigenic E. coli (ETEC) or with low-virulence TGE coronavirus, application of fumonisin B1 with a normal therapeutic dose of dexamethasone, and the increase of soybean protein concentration in the feed. A total of 41 weaned pigs served as negative controls inoculated with a commensal porcine E. coli. Housing conditions simulated the farm environment. As an overall result, ileal segments of 18.5% of infected pigs were shown to be colonised by EPEC, while no EPEC were detected in the ilea of controls. Among predisposing factors occurring on farms, feed protein content increased by 20% (26.3% crude protein, provided by 48% soybean meal) seemed to enhance EPEC colonisation and resulted in the mobilisation of spontaneous latent EPEC/ETEC infection. The results indicate that under normal farm conditions porcine EPEC may colonise conventional weaned pigs by inducing ileal attaching effacing (AE) lesions with reasonable frequency, without clinical signs. The results also suggest that conventional weaned pigs may represent undetected reservoirs of porcine EPEC, potentially giving rise to the emergence of new types of EHEC due to natural transduction by Stx phages.     

Microarray based comparative genotyping of gentamicin resistant Escherichia coli strains from food animals and humans

Recent data from the European and Hungarian Antimicrobial Resistance Monitoring Systems have indicated that the routine use of gentamicin in human and veterinary medicine frequently leads to the selection of gentamicin resistance in Escherichia coli. The aim of this study was to provide molecular characterization of gentamicin resistance in clinical and commensal E. coli strains representing humans and food producing animals by genotyping for antimicrobial resistance and virulence using a miniaturized microarray. All 50 strains tested proved to be multidrug resistant defined as resistance to three or more antimicrobial classes. Antimicrobial resistances genes such as aadA1-like, strB, bla(TEM), sul1 and tet(A) or tet(B), and corresponding phenotypes (streptomycin-, ampicillin-, sulfamethoxazole- and tetracycline resistance) were detected in >50% of isolates regardless of the host or clinical background. However, certain genes encoding gentamicin resistance such as aac(6')-Ib and ant(2″)-Ia as well as catB3-like genes for phenicol resistance were only detected in human isolates. Among virulence genes, the increased serum survival gene iss was predominant in all host groups. Although the majority of gentamicin resistant E. coli strains were characterized by diverse antimicrobial resistance, and virulence gene patterns, accentuated links between catB3-like, aac(6')-Ib, bla(CTX-M-1) and sat genes could be detected in human strains. Further resistance/virulence gene associations (tet(A) with iroN and iss) were detected in poultry strains. In conclusion, the simultaneous characterization of antimicrobial resistance and virulence genotypes of representative clinical and commensal strains of E. coli should be useful for the identification of emerging genotypes with human and or animal health implications.     

Monitoring the circadian rhythm of serum and salivary cortisol concentrations in the horse

Daily fluctuations of cortisol concentration in the blood or saliva have been repeatedly reported. However, several contradictions in the existing literature appear on this subject. The present study was performed to definitively establish options for testing adrenocortical function. To the best of our knowledge, this is the first study to evaluate parallel circadian rhythms in salivary and serum cortisol concentrations during a 24-h period. Twenty horses were examined under the same conditions. Blood and saliva samples were taken every 2 h for 24 h to determine the daily changes in cortisol concentrations of saliva and serum at rest and to determine the relationship between salivary and serum cortisol levels. Cosinor analysis of group mean data confirmed a significant circadian component for both serum and salivary cortisol concentrations (P < 0.001 in both cases). The serum cortisol circadian rhythm had an acrophase at 10:50 AM (95% CI, 10:00 AM–11:40 AM), a MESOR of 22.67 ng/mL, and an amplitude of 11.93 ng/mL. The salivary cortisol circadian rhythm had an acrophase at 10:00 AM (95% CI, 9:00 AM–11:00 AM), a MESOR of 0.52 ng/mL, and an amplitude of 0.12 ng/mL. We found a significant but weak association between salivary and serum cortisol concentrations; the Pearson correlation coefficient was 0.32 (P < 0.001). The use of salivary cortisol level as an indicator of hypothalamic-pituitary-adrenal axis activity may be warranted. However, the salivary cortisol levels are more likely to be correlated with free plasma cortisol than with the total plasma cortisol concentration.     

Predictive relationship between polyphenol and nonfat cocoa solids content of chocolate

Chocolate is often labeled with percent cocoa solids content. It is assumed that higher cocoa solids contents are indicative of higher polyphenol concentrations, which have potential health benefits. However, cocoa solids include polyphenol-free cocoa butter and polyphenol-rich nonfat cocoa solids (NFCS). In this study the strength of the relationship between NFCS content (estimated by theobromine as a proxy) and polyphenol content was tested in chocolate samples with labeled cocoa solids contents in the range of 20-100%, grouped as dark (n = 46), milk (n = 8), and those chocolates containing inclusions such as wafers or nuts (n = 15). The relationship was calculated with regard to both total polyphenol content and individual polyphenols. In dark chocolates, NFCS is linearly related to total polyphenols (r2 = 0.73). Total polyphenol content appears to be systematically slightly higher for milk chocolates than estimated by the dark chocolate model, whereas for chocolates containing other ingredients, the estimates fall close to or slightly below the model results. This shows that extra components such as milk, wafers, or nuts might influence the measurements of both theobromine and polyphenol contents. For each of the six main polyphenols (as well as their sum), the relationship with the estimated NFCS was much lower than for total polyphenols (r2 < 0.40), but these relationships were independent of the nature of the chocolate type, indicating that they might still have some predictive capabilities.     

Salsolinol is present in the bovine posterior pituitary gland and stimulates the release of prolactin both in vivo and in vitro in ruminants

The aims of the present study were to determine whether salsolinol (SAL), a dopamine-related compound, is present in the bovine posterior pituitary (PP) gland, and to clarify the effect of SAL on the secretion of prolactin (PRL) in ruminants. SAL was detected in extract of bovine PP gland using high-pressure liquid chromatography with electrochemical detection (HPLC-EC). A single intravenous (i.v.) injection of SAL (5 and 10mg/kg body weight) significantly and dose-dependently stimulated the release of PRL in goats (P<0.05). Plasma PRL levels reached a peak 10min after the injection, then gradually returned to basal values in 60-80min. The PRL-releasing pattern was similar to that in response to sulpiride (a dopamine receptor antagonist). The intracerebroventricular (i.c.v.) injection of 1mg of SAL had no significant effect on the release of PRL in calves, however, 5mg significantly stimulated the release (P<0.05) with peak values reached 30-40min after the injection. Moreover, SAL significantly stimulated the release of PRL from cultured bovine anterior pituitary cells at doses of 10(-6) and 10(-5)M, compared to control cells (P<0.05). Taken together, our data clearly show that SAL is present in extract of the PP gland of ruminants, and has PRL-releasing activity both in vivo and in vitro. Therefore, this endogenous compound is a strong candidate for the factor having PRL-releasing activity that has been previously detected in extract of the bovine PP gland.     

Long-established and emerging pesticide poisoning in horses

Poisoning is a frequent problem in domestic animals. Horses are the third most commonly affected species and pesticides are one of the main causes of toxicosis. Here within are reviewed the most frequent toxicoses where it emerges that the main causes are misuse and accidental exposure. Rodenticides, acetylcholinesterase insecticides followed by herbicides are the main cause of toxicosis and effects of emerging chemicals are being reported. Metaldehyde is still a problem, together with methiocarb. No information has been reported on poisoning with pesticide mixtures.     

Investigation of field outbreaks of turkey haemorrhagic enteritis in Hungary

Epidemiological, pathological, serological and virological investigations are reported on turkey haemorrhagic enteritis virus (THEV) infection in Hungarian turkey flocks. The pathogenesis of infection in experimentally infected turkeys and chickens, as well as the usefulness of polymerase chain reaction (PCR)/sequencing method for epidemiological investigation and for the differentiation of vaccine and field strains of THEV was also studied. Since the first recognition of the disease in Hungary in the late 1970s, until recently the disease has been diagnosed sporadically in its mild form. In the last few years (2000-2005), however, the number of outbreaks and the severity of the disease increased (9-23 affected flocks/year). Most of the outbreaks occurred at the age of 6 to 8 weeks and was complicated with Escherichia coli infection. The antibody levels to THEV in turkey flocks gradually declined till 5-7 weeks of age, and then they increased sharply due to natural infection with THEV. The immune response to vaccination (at 5 weeks of age) showed no significant antibody level increase one week postvaccination, but four weeks later the antibody level reached high values and then remained at this high level. The agar gel immunodiffusion (AGID) test to detect turkey adenovirus A (TAdV-A) antigen and PCR methods for THEV-specific DNA gave similarly positive results if spleens with pathognomonic lesions were tested; however, PCR proved to be more sensitive in cases with less characteristic pathological lesions. Nucleotide sequence alignment of PCR products amplified from Hungarian field strains and the Domermuth vaccine strain and that of the published THEV hexon sequences in GenBank database revealed slight differences between the sequences.     

Effects of elevated CO2 concentration on the respiratory behavior of the branches of field-grown hinoki cypress (Chamaecyparis obtusa)

The effects of elevated atmospheric CO₂ concentrations on the nighttime respiration were examined for two sample branches of a hinoki cypress tree (Chamaecyparis obtusa) growing in the field with an open gas exchange system for a one-year period from July 1994 to June 1995. The branches were of a similar size and located at a similar position within the crown. One branch was subjected to an elevated CO₂ concentration of 800 μmol mol⁻¹ and the other was subjected to ambient air which had a CO₂ concentration of about 370 μmol mol⁻¹. Nighttime respiration rate was higher in elevated CO₂ level than in ambient CO₂ level. The relationship between nighttime respiration and the corresponding nighttime air temperature was fitted by the exponential function in every month of the year. The segregation of regression lines between the two CO₂ treatments increased gradually as the seasons progressed during the treatment period. TheQ ₁₀ values for nighttime respiration were lower in elevated CO₂ (1.9 ≤Q ₁₀ ≤ 3.7) than in ambient CO₂ (2.4 ≤Q ₁₀ ≤ 4.5) in every month of the year. TheQ ₁₀ was inversely related to the monthly mean nighttime air temperature in both elevated and ambient CO₂. The estimated daily nighttime respiration rate under both CO₂ treatments had a similar seasonal pattern, which almost synchronized with the temperature change. The respiration ratio of elevated CO₂ to ambient CO₂ increased gradually from 1.1 to 1.6 until the end of the experiment. Our results indicate that the CO₂ level and the temperature have a strong interactive effect on respiration and suggest that a potential increase in respiration of branches will occur when ambient CO₂ increases.