A newSalmonella type (S. ngozi) isolated separately in its monophasic and biphasic form from a dog and a gecko

Summary A newSalmonella type, which was first isolated from a dog as a monophasic strain (48: z₁₀;−) and some months later from a gecko as a biphasic strain (48: z₁₀; 1.5) is described.     

Three newSalmonella types (S. Bilthoven,S. wassenaar andS. bonaire

Summary Two newSalmonella types,S. bilthoven, 47ac: a:—andS. wassenaar, 50; gp—were isolated from reptiles in the Netherlands. A newSalmonella type,S. bonaire 50; z₄z₃₂:— was isolated from a cow on the Island Bonaire.     

Six newSalmonella types,isolated in ghana (S. volta,S. agona,S. wa,S. techimani,S. mampong andS. tafo)

Summary Six newSalmonella types isolated in Ghana are described.S. volta, 11: 4: 1,z₁₃, z₂₈ was isolated from a swine;S.agona 4,12: fgs:—,S.wa, 16: b: 1,5S.technimani, 28: c: z₆ andS.tafo, 1, 4 12, 27: z₃₅: 1,7 were isolated from cattle;S.mampong, 13,22: z₃₅: 1,6, was isolated from a lizzard.     

A newSalmonella type (S. Benguella)

Summary A newSalmonella typeS. benguella, with the antigenic formula 40: b:z ₆, is described. The culture was isolated together withS. meleagridis from fish-meal of Portuguese Angola origin.     

Prevalence and numbers of Salmonella spp. and Enterobacteriaceae on pork cuts in abattoirs in the Republic of Ireland

Aims: This study aimed to determine the numbers and types of Salmonella spp. and Enterobacteriaceae on pork cuts in the meat cutting room environment of four commercial pork abattoirs in the Republic of Ireland. Methods and Results: Pork oysters (M. gluteus medius; n = 720) and swabs (n = 56) from equipment and surfaces were screened for Salmonella spp. using a DNA-based PCR method and confirmed by culture. Salmonella numbers were assessed using a three-tube most probable number (MPN) technique. Salmonella spp. was detected on 24/720 (3·3%) pork cuts (range of <0·03–0·36 MPN g⁻¹) and in 7/56 (12·5%) environmental swabs (range of <0·03–1·10 MPN cm⁻²). There was significant variation in the prevalence of Salmonella on pork between different abattoirs and days of sampling (range of 0–31·7%). The predominant serotype was Salmonella serotype Typhimurium followed by Salmonella serotype Derby. Conclusions: Overall prevalence data conceal the key finding that there was considerable variation in the incidence of Salmonella on different days. A direct association between Salmonella contamination of pork cuts and equipment/surfaces was observed. Significance and Impact of the Study: Prevalence and numbers of Salmonella were low; however, results clearly demonstrate the potential for cross-contamination from equipment and meat contact surfaces in the cutting room environment.     

Effects of nitric oxide in 5-hydroxytryptamine-, cholera toxin-, enterotoxigenic Escherichia coli- and Salmonella Typhimurium-induced secretion in the porcine small intestine

The effects of nitric oxide (NO) in the secretory response to the endogenous secretagogue 5-hydroxytryptamine (5-HT), the enterotoxins heat-labile enterotoxigenic Escherichia coli (ETEC) toxin (LT) and cholera toxin (CT), and various cultures of ETEC and Salmonella serotype Typhimurium in the porcine small intestine (Sus scrofa) were investigated. In anaesthetized pigs, jejunal tied-off loops were instilled with 5-HT, LT, CT, various cultures of ETEC or S. Typhimurium. Pigs were given intravenously isotonic saline or isotonic saline containing the NO synthase inhibitor, N^ω-nitro-l-arginine methyl ester (L-NAME). L-NAME significantly induced an increased fluid accumulation in loops induced by 5-HT, ETEC and stn-mutated S. Typhimurium. Fluid accumulation in loops instilled with wild-type S. Typhimurium was increased by L-NAME, although not significantly, while there was no effect on fluid accumulation induced by an invH-mutated isogenic strain. No significant effect of L-NAME was observed on the fluid accumulation induced by the purified enterotoxins LT and CT. The results also demonstrated a relatively large difference in the ability to induce fluid accumulation between the bacteria strains. Diastolic, systolic and mean blood pressures were significantly increased and the body temperature was significantly decreased in groups of pigs treated with L-NAME. In conclusion, the results suggest that NO has a proabsorptive effect in the intact porcine jejunum and is involved in the systemic vascular tone.     

Campylobacter,salmonella and chlamydia in free-living birds of Croatia

Campylobacteriosis, salmonellosis and avian chlamydiosis are zoonotic diseases in which birds have been suggested to play an important role as reservoirs. We have investigated the prevalence of Campylobacter and Salmonella spp. and Chlamydophila sp. in 107 free-living birds belonging to 25 species from 13 families from Croatia in order to examine the natural infections caused by these agents. Campylobacter jejuni-like organisms were isolated from 2 of 107 free-living bird species examined (1.9%). Salmonella was isolated from 8 fresh fecal specimens from free-living bird species (7.4%). These isolates were identified as S. typhimurium in 4 (3.7%), and S. enteriditis in 4 (3.7%) free-living birds. These samples originated from feral pigeons (Columba livia domesticus; n=14; 28.6%), rook (Corvus frugilegus; n=13; 15.4%), buzzard (Buteo buteo; n=12; 16.7%), black-headed gull (Larus ridibundus; n=8; 12.5%) and tawny owl (Strix aluco; n=8; 12.5%). The presence of Chlamydophila sp. was not detected in the free-living birds examined during this study. Epidemiological aspects and possible significance of the examined birds as a source of infections for domestic animals and humans are discussed.     

Protein engineering studies of A-chain loop 47-56 of Escherichia coli heat-labile enterotoxin point to a prominent role of this loop for cytotoxicity

Heat-labile enterotoxin (LT), produced by enterotoxigenic Escherichia coli, is a close relative of cholera toxin (CT). These two toxins share approximately 80% sequence identity, and consists of one 240-residue A chain and five 103-residue B subunits. The B pentamer is responsible for G_M1 receptor recognition, whereas the A subunit carries out an ADP-ribosylation of an arginine residue in the G protein, G_Sα, in the epithelial target cell. This paper explores the importance of specific amino acids in loop 47–56 of the A subunit. This loop was observed to be highly mobile in the inactive R7K mutant of the A subunit. The position of the loop in wild-type protein is such that it might require considerable reorganization during substrate binding and is likely to have a crucial role in substrate binding. Five single-site substitutions have been made in the LT-A subunit 47–56 loop to investigate its possible role in the enzymatic activity and toxicity of LT and CT. The wild-type residues Thr-50 and Val-53 were replaced either by a glycine or by a proline. The glycine substitutions were intended to increase the mobility of this active-site loop, and the proline substitutions were intended to decrease the mobility of this same loop by restricting the accessible conformational space. Under the hypothesis that mobility of the loop is important for catalysis, the glycine-substitution mutants T50G and V53G would be expected to exhibit activity equal to or greater than that of the wild-type A subunit, while the proline substitution mutants T50P and T53P would be less active. Cytotoxicity assays showed, however, that all four of these mutants were considerably less active than wild-type LT. These results lend support for assignment of a prominent role to loop 47–56 in catalysis by LT and CT.     

Testing the hypothesis on the relationship between aerodynamic roughness length and albedo using vegetation structure parameters

Surface albedo (α) and aerodynamic roughness length (z ₀), which partition surface net radiation into energy fluxes, are critical land surface properties for biosphere–atmosphere interactions and climate variability. Previous studies suggested that canopy structure parameters influence both α and z ₀; however, no field data have been reported to quantify their relationships. Here, we hypothesize that a functional relationship between α and z ₀ exists for a vegetated surface, since both land surface parameters can be conceptually related to the characteristics of canopy structure. We test this hypothesis by using the observed data collected from 50 site-years of field measurements from sites worldwide covering various vegetated surfaces. On the basis of these data, a negative linear relationship between α and log(z ₀) was found, which is related to the canopy structural parameter. We believe that our finding is a big step toward the estimation of z ₀ with high accuracy. This can be used, for example, in the parameterization of land properties and the observation of z ₀ using satellite remote sensing.     

Dombrock blood group (DO): assignment to chromosome 12p

The Dombrock blood group system (DO) is a common polymorphism in Caucasians, represented by two red cell antigen alleles. In a linkage study in our family material of 832 families from the Copenhagen area, we found a strong indication of tight linkage with the two flanking DNA polymorphisms D12S358 (z = 7.66; at θ _M = 0.001, θ _F = 0.031) and D12S364 (z = 8.53; at θ _M = 0.068, θ _F = 0.031). DO is assigned to the region 12p13.2– 12p12.1 by physically localised markers. Received: 18 April 1996 / Revised: 4 July 1996     

Apparent charge of binding site in ion-translocating enzymes: kinetic impact

Recently, we presented a general scope for the nonlinear electrical properties of enzymes E which catalyze translocation of a substrate S with charge number z_S through lipid membranes (Boyd et al. J. Membr. Biol. 195:1–12, 2003). In this study, the voltage sensitivity of the enzymatic reaction cycle has been assigned to one predominant reversible reaction step, i.e. the reorientation of either E or ES in the electric field, leaving the reorientation of the alternate state (ES or E) electroneutral, respectively. With this simplification, the steady-state current–voltage relationships (IV) assumed saturation kinetics like in Michaelis–Menten systems. Here, we introduce an apparent charge number z_E of the unoccupied binding site of the enzyme, which accounts for the impact of all charged residues in the vicinity of the physical binding site. With this more realistic concept, the occupied binding site assumes an apparent charge of , and IV does not saturate any more in general, but exponentially approaches infinite or zero current for large voltage displacements from equilibrium. These nonlinear characteristics are presented here explicitly. They are qualtitatively explained in a mechanistic way, and are illustrated by simple examples. We also demonstrate that the correct determination of the model parameters from experimental data is still possible after incorporating z_E and its corollaries into the previous model of enzyme-mediated ion translocation.     

Frequency of indicator bacteria,Salmonella and diarrhoeagenic Escherichia coli pathotypes on ready‐to‐eat cooked vegetable salads from Mexican restaurants

The presence of coliform bacteria, faecal coliforms, Escherichia coli, diarrhoeagenic E. coli pathotypes (DEP) and Salmonella were determined in ready‐to‐eat cooked vegetable salads (RECS) from restaurants in Pachuca city, Mexico. The RECS were purchased from three types of restaurants: national chain restaurants (A), local restaurants (B) and small restaurants (C). Two restaurants for each A and B, and three for C, were included. Forty RECS samples were purchased at each A and B restaurant and 20 at each C restaurant. Of the overall total of 220 analysed samples, 100, 98·2, 72·3, 4·1 and 4·1% had coliform bacteria, faecal coliforms, E. coli, DEP and Salmonella, respectively. Identified DEP included enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC) and Shiga toxin‐producing E. coli (STEC). The EPEC, ETEC and STEC were isolated each from 1·4% of samples. No E. coli O157:H7 were detected in any STEC‐positive samples. The analysis of Kruskal–Wallis anova and median test of microbiological data showed that the microbiological quality of RECS did not differ between the different restaurants (P > 0·05).

Significance and Impact of the Study

This is the first report regarding microbiological quality and Salmonella, enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC) and Shiga toxin‐producing E. coli (STEC) isolation from ready‐to‐eat cooked vegetable salads from Mexican restaurants. Ready‐to‐eat cooked vegetable salads could be an important factor contributing to the endemicity of EPEC, ETEC and STEC, and Salmonella caused gastroenteritis in Mexico.     

Bending of z-lines by mechanical stimuli: An input signal for integrin dependent modulation of ion channels?

We studied which components of mechanical cell deformation are involved in “stretch modulated ion currents” (SMIC). Murine ventricular myocytes were attached to glass coverslips and deformed in x, y and z with a 16 μm thin glass stylus (S) of calibrated stiffness. Three-dimensional confocal microscopy characterized cell deformation (T-tubular membranes, mitochondria) and bending of S (indicative of the applied force). Axial (x-) displacement of S sheared the upper cell part versus the attached bottom, close to S, it changed sarcomere length and bent z-lines (“z-line displacement”). Vertical (z-press) or transversal (y-shear) displacement of S bulged cytoplasm and mitochondria transversally without detectable z-line displacement.Axial stiffness increased with the extent of stress (“stress stiffening”). Depolymerization of F-actin or block of integrin receptors reduced stiffness. SMIC served as a proxy readout of deformation-induced signaling. Axial deformation activated a non-selective cation conductance (G_ns) and deactivated an inwardly rectifying K⁺ conductance (G_K1), z-press or y-shear did not induce SMIC. Depolymerization of F-actin or block of integrin receptors reduced SMIC. SMIC did not depend on changes in sarcomere length but correlated with the extent of z-line bending. We discuss that both shear stress at the attached cell bottom and z-line bending could activate mechanosensors. Since SMIC was absent during deformations without z-line bending we postulate that z-line bending is a necessary component for SMIC signaling.     

Eight newSalmonella types

Summary Eight newSalmonella types are described.S. houten, 43; Z₄, Z₂₃ :−, was isolated from a frog,S.tuindorp, 43: Z₄, Z₃₂: −,S. vleuten, 44: fg: −,S.zeist, 18: Z₁₀: Z₆, andS.soesterberg. 21: Z₄, Z₂₃: −, were isolated from lizards. S. kralingen (8), 20: y: Z₆, was isolated from peanuts andS. assen, 21: a: −, was isolated from a swine. All these strains were isolated in the Netherlands. S.mim, 13, 22: a: 1,6, was isolated from a rat in Ghana.     

Salmonella enterica isolates from pasture‐raised poultry exhibit antimicrobial resistance and class I integrons

Aims: While considerable foodborne pathogen research has been conducted on conventionally produced broilers and turkeys, few studies have focused on free‐range (organic) or pastured poultry. The current surveillance study was designed to isolate, identify and genetically characterize Salmonella from pastured poultry farm environment and from retail samples. Methods and Results: In this study, 59 isolates were collected from two pastured poultry farms (n = 164; pens, feed, water and insect traps) and retail carcasses (n = 36) from a local natural foods store and a local processing plant. All isolates were serotyped and analysed phenotypically (antimicrobial resistance profiles) and genotypically (DNA fingerprints, plasmid profiles and integron analysis). Salmonella enterica was detected using standard microbiological methods. Salmonella Kentucky was the most prevalent serotype detected from the sampled sources (53%), followed by Salmonella Enteritidis (24%), Bareilly (10%), Mbandaka (7%), Montevideo (5%) or Newport (2%). All isolates were resistant to sulfisoxazole and novobiocin, and the majority (40/59) possessed class I integrons shown by PCR detection. Each Salmonella serotype elicited a distinct pulsed‐field gel electrophoresis fingerprint profile, and unique differences were observed among the serotypes. Conclusions: The findings of this study show that Salmonella serotypes isolated from pasture‐raised poultry exhibit antimicrobial resistance and class I integrons. Significance and Impact of the Study: This study demonstrates that despite the cessation of antibiotic usage in poultry production, antibiotic resistant Salmonella may still be recovered from the environment and poultry products.     

Salmonella spp. are affected by different levels of water activity in closed microcosms

Controlling water activity (a _w) can significantly impact the growth of Salmonella in poultry litter and manure — a phenomenon that was studied quantitatively using two common serotypes of Salmonella. The quantitative effect of changes in levels of a _w on Salmonella populations was determined using inoculated, frosted glass rectangles placed in closed chambers (microcosms). Glass rectangles with known concentrations of Salmonella enteritidis and S. brandenburg were placed in microcosms maintained at an a _w level of 0.893 for 24 h at room temperature (RT) and then transferred to other microcosms maintained at the same temperature but with higher a _w levels (0.932 and 0.987). Salmonella populations on the slides were quantified at 4, 18, 24, and 48 h. Slightly elevated levels of a _w (<0.1, i.e., 10% equilibrium relative humidity) for 24 h resulted in a 100-fold increase in counts of Salmonella. The data also suggested that in vitro adaptation to dry environments may occur when the organisms are exposed to alternating levels of relatively high and low (0.987 and 0.893) levels of a _w. Any increased tolerance of Salmonella to reduced levels of a _w could be the result of physico-chemical changes in the organism due to selective environmental pressure, formation of a protective biofilm, and/or entry into a dormant state. Results from this study are compatible with those from previously reported on-farm surveys, reinforcing the contention that maintaining a _w below 0.85 in and around litter/manure surfaces in poultry or livestock bedding areas may be a critical factor in safe production of food. Journal of Industrial Microbiology & Biotechnology (2001) 26, 222–225. Received 18 May 2000/ Accepted in revised form 24 January 2001     

Design of a Prediction Model for the Differentiation of Pasteurized Milk from Heated ESL Milk by Peptide Profiling

This study designs a prediction model to differentiate pasteurized milk from heated extended shelf life (ESL) milk based on milk peptides. For this purpose, quantitative peptide profiles of a training set of commercial samples including pasteurized (n = 20), pasteurized‐ESL (n = 13), and heated‐ESL (n = 16) milk are recorded by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF‐MS). Seven peptides are selected as putative markers, and cutoff levels and performance measures of each marker are defined by receiver operating characteristic (ROC) analysis. The accuracy of these peptides in the training set range between 71% and 90%. A prediction model is established based on the combined cutoff levels and evaluated by an independent blind test set. The processing method of 19 out of 20 unknown milk samples is predicted correctly achieving 95% accuracy. Five peptides of the prediction model are identified as α_S1‐casein_182–199 (m/z 2014.0), α_S1‐casein_180–199 (m/z 2216.1), α_S1‐casein_1–24 (m/z 2910.6), β‐casein_108–125 (m/z 2126.0), and β‐casein_106–125 (m/z 2391.2) indicating thermal release and the action of plasmin and cathepsins. Thus, the present study demonstrates that the milk peptide profile reflects even minor differences in production parameters.     

Treatment with Ca(OH)2 for inactivation of Salmonella Typhimurium and Enterococcus faecalis in soil contaminated with infected horse manure

Aim: To investigate the inactivation of Salmonella enterica serovar Typhimurium and the faecal indicator Enterococcus faecalis in horse manure:soil mixtures by application of hydrated lime (Ca(OH)₂). Methods and Results: In laboratory incubations, the inhibitory effect of different concentrations of Ca(OH)₂, as well as different application techniques, was tested. Other variables were horse manure:soil ratio, incubation temperature (6 and 14°C) and soil type (sand/clay). Bacterial enumeration by the plate count method in samples taken at increasing intervals revealed that Ca(OH)₂ effectively reduced Salmonella Typhimurium numbers. However, to achieve a sufficient reduction, the Ca(OH)₂ had to be applied at a sufficient rate, and the amount required varied because of manure:soil ratio and incubation temperature. The results showed that a pH above 11 was needed and that a high pH had to be maintained for up to 7 days. An appropriate application technique for the Ca(OH)₂ was also important, so that a high pH was obtained throughout the whole material to be treated. In addition, a high manure:soil ratio in combination with a higher incubation temperature was found to rapidly neutralize the pH and to increase the risk of Salmonella re‐growth. Conclusions: Application of Ca(OH)₂ can be an efficient method for treating a Salmonella‐contaminated horse paddock. A high pH is a key factor in Salmonella inactivation, and thus, monitoring the pH during the treatment period is necessary. To avoid re‐growth excess manure should be removed for separate treatment elsewhere. Significance and Impact of the Study: Persistence of Salmonella in horse paddocks poses a risk of disease transmission to healthy animals and people who come into contact with these animals. An efficient method to de‐contaminate a Salmonella‐contaminated soil would be a valuable tool for animal welfare and for public health.     

Occurrence of Salmonella spp in estuarine and coastal waters of Portugal

The presence of Salmonella and its relationship with indicator organisms of fecal pollution, such as total coliforms, fecal coliforms and fecal streptococci, was studied at two marine zones in Portugal. Seventeen different Salmonella serotypes were isolated and identified, S. virchow was the most frequently isolated (21.6%). In addition, a high percentage (35.1%) was recorded for some Salmonella serotypes of clinical significance, namely S. enteritidis, S. infantis, S. typhimurium and S. virchow. In any of the samples from the two zones Salmonella was not detected in the absence of any of the indicator organisms. However, the incidence of Salmonella as a function of indicator concentration intervals established by the EEC standards was 0, 10 and 19.3% at guide values of total coliforms, fecal coliforms and fecal streptococci, respectively in the Faro samples (south of Portugal). In contrast, Salmonella incidence rates of 37.5, 36.4 and 33.3% were recorded at the corresponding guide values the Caminha samples (north of Portugal). No significant correlations (p>0.005) were obtained between Salmonella and the indicators at the sampling stations; however, total coliforms and fecal streptococci were the indicators most closely related to Salmonella in Caminha and Faro samples, respectively. Survival experiments in Escherichia coli, Enterococcus faecalis and S. typhimurium, using diffusion chambers, were performed to verify whether the lack of correlation between indicators and Salmonella was due to different inactivation rates in seawater. The results indicate that survival percentages of the three microorganisms tested were similar after 48 h of exposure to seawater.