Characterization of multiple-antimicrobial-resistant Salmonella enterica Subsp. enterica isolated from indigenous vegetables and poultry in Malaysia

Aims: The aims of this communication were to study characterization of serogroups among Salmonella isolates and the relationship of antimicrobial resistance to serogroups. Multiple antimicrobial resistance (MAR) was performed on 189 Salmonella enterica isolates associated with 38 different serovars that were recovered from poultry and four types of indigenous vegetables. Methods and Results: Disc diffusion analysis was performed with a selection of 10 different antimicrobial agents. Isolates recovered from indigenous vegetables showed 100% (134/134) resistant to erythromycin and followed by 42%, 34%, 19% for tetracycline, streptomycin and trimethroprim‐sulfamethoxazole respectively. In general, 90·1% (50/55) and 56·7% (76/134) of Salmonella isolated from poultry and indigenous vegetables, respectively, exhibited MAR index more than 0·2. Conclusions: Characterization of Salmonella isolates based on the MAR results indicated that poultry still remains as the main reservoir for multi‐drug‐resistant Salmonella. Four isolates from the indigenous vegetables showed the highest MAR index in this study. Further investigations need to be conducted to determine if Salmonella isolates recovered from indigenous vegetables were gaining more antimicrobial resistance. Significance and Impact of the Study: The study enabled us to determine antimicrobial patterns and trends in Salmonella from poultry and indigenous vegetables in Malaysia.     

Health Risk Assessment of Consumption of Heavy Metals in Market Food Crops from Sialkot and Gujranwala Districts,Pakistan

This study was performed to investigate the potential health risk of heavy metals (HMs) through consumption of market food crops (MFCs) in the Sialkot and Gujranwala districts, Pakistan. Both study areas are located in industrialized regions of the country, where atmospheric pollution is a problem and irrigation of food crops is mostly practiced on the use of wastewater/contaminated water. For the purpose of this study, MFCs samples were collected and assessed for HMs (Cr, Ni, Cd, Pb, Mn, Cu, Zn, and Fe) by using flame atomic absorption spectrophotometry. Concentration of HMs such as Pb and Cd exceeded the Food and Agriculture/World Health Organization's recommended limits in all MFCs, while Cr in most of the vegetables of the Sialkot and Gujranwala districts also exceeded that limit. The health risk index was >1 in Triticum aestivum for Pb and Cd intake in the Sialkot district and only Pb in the Gujranwala district. Therefore, this study suggests pretreatment of wastewater and its utilization for lawns and green belts irrigation, rather than for food crops. This study also suggests a regular monitoring of HMs in the irrigation water, subsequent soil, air, and MFCs in order to prevent or reduce health hazards.     

Fate of Salmonella enterica Serovar Typhimurium on Carrots and Radishes Grown in Fields Treated with Contaminated Manure Composts or Irrigation Water

Three different types of compost, PM-5 (poultry manure compost), 338 (dairy cattle manure compost), and NVIRO-4 (alkaline-pH-stabilized dairy cattle manure compost), and irrigation water were inoculated with an avirulent strain of Salmonella enterica serovar Typhimurium at 10⁷ CFU g⁻¹ and 10⁵ CFU ml⁻¹, respectively, to determine the persistence of salmonellae in soils containing these composts, in irrigation water, and also on carrots and radishes grown in these contaminated soils. A split-plot block design plan was used for each crop, with five treatments (one without compost, three with each of the three composts, and one without compost but with contaminated water applied) and five replicates for a total of 25 plots for each crop, with each plot measuring 1.8 × 4.6 m. Salmonellae persisted for an extended period of time, with the bacteria surviving in soil samples for 203 to 231 days, and were detected after seeds were sown for 84 and 203 days on radishes and carrots, respectively. Salmonella survival was greatest in soil amended with poultry compost and least in soil containing alkaline-pH-stabilized dairy cattle manure compost. Survival profiles of Salmonella on vegetables and soil samples contaminated by irrigation water were similar to those observed when contamination occurred through compost. Hence, both contaminated manure compost and irrigation water can play an important role in contaminating soil and root vegetables with salmonellae for several months.     

High levels of microbial contamination of vegetables irrigated with wastewater by the drip method.

The public health aspects of the use of wastewater in agriculture and the effects of the drip irrigation method on the contamination of vegetables were studied. The method used was to simulate enteric microorganisms' dissemination by contaminated irrigation water in the field. The vegetables were irrigated with an effluent inoculated with a high titer of traceable microorganisms: poliovirus vaccine and a drug-resistant Escherichia coli. The dissemination of the marker organisms in the field was followed, and the effects of certain manipulations of the drip irrigation method on the contamination of the crops by the effluent were examined. It was shown that drip irrigation under plastic sheet cover with the drip lines placed either on the soil surface or buried at a depth of 10 cm significantly reduced crop contamination from inoculated irrigation water even when massive doses of bacteria and viruses were used. The microbial contamination was found to persist in the irrigation pipes and in the soil for at least 8 and 18 days, respectively. The data indicate that the recovery of the marker organisms was affected by soil texture and environmental conditions.     

High levels of microbial contamination of vegetables irrigated with wastewater by the drip method.

The public health aspects of the use of wastewater in agriculture and the effects of the drip irrigation method on the contamination of vegetables were studied. The method used was to simulate enteric microorganisms' dissemination by contaminated irrigation water in the field. The vegetables were irrigated with an effluent inoculated with a high titer of traceable microorganisms: poliovirus vaccine and a drug-resistant Escherichia coli. The dissemination of the marker organisms in the field was followed, and the effects of certain manipulations of the drip irrigation method on the contamination of the crops by the effluent were examined. It was shown that drip irrigation under plastic sheet cover with the drip lines placed either on the soil surface or buried at a depth of 10 cm significantly reduced crop contamination from inoculated irrigation water even when massive doses of bacteria and viruses were used. The microbial contamination was found to persist in the irrigation pipes and in the soil for at least 8 and 18 days, respectively. The data indicate that the recovery of the marker organisms was affected by soil texture and environmental conditions.     

Fresh fruit and vegetables as vehicles for the transmission of human pathogens

Much research into food‐borne human pathogens has focused on transmission from foods of animal origin. However, recent investigations have identified fruits and vegetables are the source of many disease outbreaks. Now believed to be a much larger contributor to produce‐associated outbreaks than previously reported, norovirus outbreaks are commonly caused by contamination of foods from hands of infected workers. Although infections with Shiga toxin‐producing E. coli O157 have been linked to beef more often than to any other food product, severe outbreaks have been traced to consumption of contaminated radish sprouts and pre‐packaged spinach. Similarly, while infections with Salmonella have mainly been linked to consumption of foods of animal origin, many outbreaks have been traced to contaminated fresh produce. E. coli O157 binds to lettuce leaves by alternative mechanisms involving the filamentous type III secretions system, flagella and the pilus curli. Association of Salmonella with fresh produce appears to be serovar‐specific involving flagella, curli, cellulose, and O antigen capsule. A better understanding of plant, microbiological, environmental, processing and food handling factors that facilitate contamination will allow development of evidence‐based policies, procedures and technologies aimed at reducing the risk of contamination of fresh produce.     

Heavy metal phyto-accumulation in leafy vegetables irrigated with municipal wastewater and human health risk repercussions

In this study, heavy metal phyto-accumulation potential of selected cultivars of two leafy vegetables on irrigation with municipal wastewater and human health risks were investigated. Municipal wastewater chemistry was recorded significantly different from groundwater control and led to the two-fold high enrichment of soil heavy metal contents (Ni, 19.46; Pb, 23.94; Co, 4.68; Cd, 1.4 in mg/kg, respectively). Interactive effects for phyto-accumulation of most heavy metals were also recorded significant at p?相似文献   

A Large-Scale Community-Based Outbreak of Paratyphoid Fever Caused by Hospital-Derived Transmission in Southern China

Background

Since the 1990s, paratyphoid fever caused by Salmonella Paratyphi A has emerged in Southeast Asia and China. In 2010, a large-scale outbreak involving 601 cases of paratyphoid fever occurred in the whole of Yuanjiang county in China. Epidemiological and laboratory investigations were conducted to determine the etiology, source and transmission factors of the outbreak.

Methodology/Principal Findings

A case-control study was performed to identify the risk factors for this paratyphoid outbreak. Cases were identified as patients with blood culture–confirmed S. Paratyphi A infection. Controls were healthy persons without fever within the past month and matched to cases by age, gender and geography. Pulsed-field gel electrophoresis and whole-genome sequencing of the S. Paratyphi A strains isolated from patients and environmental sources were performed to facilitate transmission analysis and source tracking. We found that farmers and young adults were the populations mainly affected in this outbreak, and the consumption of raw vegetables was the main risk factor associated with paratyphoid fever. Molecular subtyping and genome sequencing of S. Paratyphi A isolates recovered from improperly disinfected hospital wastewater showed indistinguishable patterns matching most of the isolates from the cases. An investigation showed that hospital wastewater mixed with surface water was used for crop irrigation, promoting a cycle of contamination. After prohibition of the planting of vegetables in contaminated fields and the thorough disinfection of hospital wastewater, the outbreak subsided. Further analysis of the isolates indicated that the origin of the outbreak was most likely from patients outside Yuanjiang county.

Conclusions

This outbreak is an example of the combined effect of social behaviors, prevailing ecological conditions and improper disinfection of hospital wastewater on facilitating a sustained epidemic of paratyphoid fever. This study underscores the critical need for strict treatment measures of hospital wastewater and the maintenance of independent agricultural irrigation systems in rural areas.     

Potential human health risk assessment of cylindrospermopsin accumulation and depuration in lettuce and arugula

The cyanobacterial toxin cylindrospermopsin (CYN) has become a globally important secondary metabolite due to the negative effect it has on human and animal health. As a means of evaluating the risk of human exposure to CYN, the bioaccumulation and depuration of the toxin in lettuce (Lactuca sativa L.) and arugula (Eruca sativa Mill.) were investigated, after irrigation with contaminated water. The vegetables were irrigated for 7 days with CYN (3, 5 and 10 μg/L) contaminated water (bioaccumulation phase), and subsequently, irrigated for 7 days with uncontaminated distilled water (depuration phase). In general, the bioaccumulation of CYN in both vegetables decreased with increasing exposure concentration. Bioconcentration factor (BCF) of CYN increased with the progression of the experiment at 3.0 μg/L CYN, while the reverse occurred at 5 and 10 μg/L CYN. In arugula, BCF increased at all CYN exposure concentrations throughout the study. The depuration of CYN decreased with increasing exposure concentration but was highest in the plants of both species with the highest bioaccumulation of CYN. Specifically, in plants previously irrigated with water contaminated with 3, 5 and 10 μg/L CYN, the depuration of the toxin was 60.68, 27.67 and 18.52% for lettuce, and 47, 46.21 and 27.67% for arugula, respectively. Human health risks assessment revealed that the consumption of approximately 10 to 40 g of vegetables per meal will expose children and adults to 1.00-6.00 ng CYN/kg body mass for lettuce and 2.22-7.70 ng CYN/kg body mass for arugula. The irrigation of lettuce and arugula with contaminated water containing low CYN concentrations constitutes a potential human exposure route.     

Wastewater as a Non-conventional Resource: Impact of Trace Metals and Bacteria on Soil,Plants, and Human Health

Abstract

In many arid and semi-arid regions, farmers are often obligated to informally use raw wastewater for irrigating their crops. The impacts of wastewater irrigation on soil, crops, and human health were investigated, regarding trace metals and bacteria. Cr, Cu, Fe, Ni, and Zn were detected in wastewater. Cr, Cu, and Zn accumulated in soil and crops in the order rocket?>?clover?>?cabbage. The Health Risk Index reported risk from Cr and Zn in rocket. Fecal coliforms in wastewater and crops were detected along with Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa. The frequency (%) was 86.7% (cabbage), 66.7% (rocket), and 43.3% (clover). The multiple antibiotic resistance index (0.503) rendered crops high risk source for contamination. The comprehensive pollution index classified wastewater severely polluted (≥2.01). Conclusions deduced crops alternative reservoirs for trace metals and human pathogens. Recommendations included implementation of low cost treatment methods, holding irrigation 5–15?days before harvest, assuming citric and acetic acids reducing solutions for trace metals, and suggested ofloxacin, amoxycillin/clavulanate, and amikacin antibiotics against reported bacteria.     

Endophytic fungi occurring in fennel, lettuce, chicory, and celery — commercial crops in southern Italy

The occurrence of endophytic fungi in fennel, lettuce, chicory, and celery crops was investigated in southern Italy. A total of 186 symptomless plants was randomly collected and sampled at the stage of commercial ripeness. Fungal species of Acremonium, Alternaria, Fusarium, and Plectosporium were detected in all four crops; Plectosporium tabacinum was the most common in all crop species and surveyed sites. The effect of eight endophytic isolates (five belonging to Plectosporium tabacinum and three to three species of Acremonium) inoculated on lettuce plants grown in gnotobiosis was assessed by recording plant height, root length and dry weight, collar diameter, root necrosis, and leaf yellowing. P. tabacinum and three species of Acremonium, inoculated on gnotobiotically grown lettuce plants, showed pathogenic activity that varied with the fungal isolate. Lettuce plants inoculated with the isolates Ak of Acremonium kiliense, Ac of Acremonium cucurbitacearum, and P35 of P. tabacinum showed an increased root growth, compared to the non-inoculated control. The high frequency of P. tabacinum isolation recorded in lettuce plants collected in Bari and Metaponto, and in fennel plants from Foggia agricultural districts, suggests a relationship not only between a crop species and P. tabacinum, but also between the occurrence of the endophyte and the crop rotation history of the soil.     

Do leafy green vegetables and their ready-to-eat [RTE] salads carry a risk of foodborne pathogens?

Over the past 10 years, there is an increasing demand for leafy green vegetables and their ready-to-eat (RTE) salads since people changed their eating habits because of healthier lifestyle interest. Nevertheless fresh leafy green vegetables and their RTE salads are recognized as a source of food poisoning outbreaks in many parts of the world. However, this increased proportion of outbreaks cannot be completely explained by increased consumption and enhanced surveillance of them. Both in Europe and in the USA, recent foodborne illness outbreaks have revealed links between some pathogens and some leafy green vegetables such as mostly lettuces and spinaches and their RTE salads since fresh leafy green vegetables carry the potential risk of microbiological contamination due to the usage of untreated irrigation water, inappropriate organic fertilizers, wildlife or other sources that can occur anywhere from the farm to the fork such as failure during harvesting, handling, processing and packaging. Among a wide range of pathogens causing foodborne illnesses, Escherichia coli O157:H7, Salmonella spp., and Listeria monocytogenes are the most common pathogens that contaminate leafy green vegetables. Children, the elderly, pregnant women and immunocompromised people are the most at risk for developing complications from foodborne illness as a result of eating contaminated leafy greens or their RTE salads. These outbreaks are mostly restaurant associated or they sometimes spread across several countries by international trade routes. This review summarizes current observations concerning the contaminated leafy green vegetables and their RTE salads as important vehicles for the transmission of some foodborne pathogens to humans.     

Bacteriological quality of crops irrigated with wastewater in the Xochimilco plots, Mexico City, Mexico

Xochimilco county plots (Mexico City), one of the most fertile agricultural areas in the Valley of Mexico, produce a large portion of the fresh vegetables consumed in the city. These plots are generally irrigated with domestic wastewater, and for this reason, it was deemed important to examine and evaluate the bacteriological quality of the water, soil, and vegetables from these plots that are harvested and marketed. The soils were also examined for the classical parameters such as nitrates, ammonia, etc., and organic matter and texture. The crops selected for this study were radishes, spinach, lettuce, parsley, and celery because they are usually consumed raw. The highest bacterial counts were encountered in leafy vegetables, i.e., spinach (8,700 for total coliform and 2,400 for fecal coliform) and lettuce (37,000 for total coliform and 3,600 for fecal coliform). Statistically significant differences in bacterial counts between rinsed and unrinsed edible portions of the crops were observed even in rinsed vegetables, and high densities of fecal coliform were detected, indicating that their consumption represents a potential health hazard. The total coliform values found in irrigation water ranged from 4 X 10(4) to 29 X 10(4), and for fecal coliform the values ranged from 5 X 10(2) to 30 X 10(2).     

Inactivation of human pathogens and spoilage bacteria on the surface and internalized within fresh produce by using a combination of ultraviolet light and hydrogen peroxide

Aims: To evaluate the efficacy of ultraviolet (UV) light (254 nm) combined with hydrogen peroxide (H₂O₂) to inactivate bacteria on and within fresh produce. Methods and Results: The produce was steep inoculated in bacterial cell suspension followed by vacuum infiltration. The inoculated samples were sprayed with H₂O₂ under constant UV illumination. The log count reduction (LCR) of Salmonella on and within lettuce was dependent on the H₂O₂ concentration, temperature and treatment time with UV intensity being less significant. By using the optimized parameters (1·5% H₂O₂ at 50°C, UV dose of 37·8 mJ cm^?2), the surface Salmonella were reduced by 4·12 ± 0·45 and internal counts by 2·84 ± 0·34 log CFU, which was significantly higher compared with H₂O₂ or UV alone. Higher LCR of Escherichia coli O157:H7, Pectobacterium carotovora, Pseudomonas fluorescens and Salmonella were achieved on leafy vegetables compared with produce, such as cauliflower. In all cases, the surface LCR were significantly higher compared with the samples treated with 200 ppm hypochlorite. UV–H₂O₂‐treated lettuce did not develop brown discolouration during storage but growth of residual survivors occurred with samples held at 25°C. Conclusions: UV–H₂O₂ reduce the bacterial populations on and within fresh produce without affecting the shelf‐life stability. Significance of the Study: UV–H₂O₂ represent an alternative to hypochlorite washes to decontaminate fresh produce.     

Epidemiology of Pseudomonas cichorii,the Cause of Lettuce Midrib Rot

Bacterial midrib rot, caused by Pseudomonas cichorii, has become a serious threat to the production of greenhouse butterhead lettuce (Lactuca sativa L. var. capitata) in Belgium. Currently, there are no strategies for controlling this pathogen. Therefore, greenhouse experiments were conducted to obtain more knowledge about the epidemiology of P. cichorii on butterhead lettuce. Greenhouse butterhead lettuce becomes susceptible to lettuce midrib rot infections at head formation, and a single overhead irrigation with water containing 10² CFU/ml P. cichorii was sufficient to cause disease. The use of surface drip irrigation instead of overhead sprinkler irrigation significantly reduced midrib rot incidence in the greenhouse. P. cichorii isolates can be divided into subgroups based on BOX‐PCR genomic fingerprinting, with isolates belonging to subgroup C1 and C2 being more virulent than those of (or related to) subgroup C3. P. cichorii infections with distinct symptoms comparable to midrib rot have also been observed on field‐grown crisphead lettuce in California and Japan which, respectively, are referred to as ‘varnish spot’ or ‘tar’. We showed that symptom expression is strongly influenced by the lettuce cultivar group, irrespective of the P. cichorii isolate, resulting in varnish spot/tar on crisphead lettuce and midrib rot on butterhead or cutting group lettuce.     

Influence of groundwater and wastewater irrigation on lead accumulation in soil and vegetables: Implications for health risk assessment and phytoremediation

The current study evaluated the effect of groundwater and wastewater irrigation on lead (Pb) accumulation in soil and vegetables, and its associated health implications. A pot experiment was conducted in which spinach (Spinacia oleracea), radish (Raphanus sativus), and cauliflower (Brassica oleracea) were irrigated with groundwater and wastewaters containing varying concentrations of Pb. Lead contents were measured in wastewaters, soils and root and shoot of vegetables. We also measured health risk index (HRI) associated with the use of vegetables irrigated by wastewaters. Results revealed that Pb contents in groundwater and wastewater samples (range: 0.18–0.31 mg/L) were below the permissible limits (0.5 mg/L) set by the Food and Agriculture Organization (FAO). Application of Pb-containing groundwater and wastewater increased Pb concentration in soil and vegetables. Lead concentrations in all soils ranged from 10 to 31 mg/kg and were below the permissible limits of 300 mg/kg set by the European Union. Significant Pb enrichment was observed in the soils whereby all types of vegetables were grown and assessed for Pb risk. Our data showed that Pb contents, in all three vegetables (21–28 mg/kg DW), were higher than the permissible Pb limit of FAO (5 mg/kg Dry Weight (DW)). The HRI values were > 1.0 for radish and cauliflower. It is proposed that Vehari city wastewater/groundwater must be treated prior to its use for irrigation to avoid vegetable contamination by Pb, and as such for reducing Pb-induced human health risk.     

The contribution of soil adhesion to radiocaesium uptake by leafy vegetables

The Goiânia accident, Brazil, was used as an opportunity to quantify the contributions of different mechanisms, in particular mass loading, leading to caesium uptake by leafy vegetables in a semi-urban environment contaminated with¹³⁷Cs. Soil splash contributions of 70–90% were quantified for lettuce and 50–60% for green cole. Soil mass loadings of 130 and 340 mg · g^–1 were estimated for lettuce and 120 and 150 mg · g^–1 for green cole. The results call attention to the potential significant contribution of the soil splash to radionuclide uptake by plants which have the edible plant parts near the soil surface (within 30–40 cm) and low root uptake factors. For radiological assessment purposes it could also be necessary to consider the contamination of crops by this mechanism.     

Salmonella grows massively and aerobically in chicken faecal matter

The use of wastewater for irrigation and animal manure as fertilizer can cause transmission of intestinal pathogens, conditions frequently observed in low- and middle-income countries (LMICs). Here, we tested the ability of Salmonella to grow in the faecal matter. We inoculated freshly isolated Salmonella strains (from chickens) in chicken faecal matter and incubated for 1 to 12 days, under aerobic and anaerobic conditions. We found that both Salmonella and Escherichia coli multiplied massively in faecal matter outside a host and significantly higher in aerobic conditions. Our results have critical implications in waste management, as we demonstrate that aerobic treatments may not be the best to reduce the number of Salmonella in the environment.     

An eight-hour PCR-based technique for detection of <Emphasis Type="Italic">Salmonella</Emphasis> serovars in seafood

A rapid and sensitive 8-h PCR assay has been developed for detection of Salmonella serovars in seafood. A total of 110 fresh and raw seafood samples were analysed for the presence of Salmonella using different enrichment periods prior to PCR assay. Seafood samples included in this study were fish, shrimps, mussels, crabs, edible oysters, and clams, collected from local fish markets in Cochin (India). The assay was performed with a Salmonella-specific 284 bp invA gene amplicon. Specificity and sensitivity of the assay were ascertained with seafoods spiked with viable Salmonella cells to a level of 10⁶ to 2 CFU per 25 g. Detection efficiency of the assay increased with increasing enrichment period for seafood, and 33.6% of seafood samples were found positive for Salmonella by 8-h PCR assay. Detection limit for the 8-h PCR assay showed visible 284 bp amplicon from seafood homogenates spiked with 2 CFU per 25 g. Seafood samples spiked with different Salmonella serovars, namely Salmonella typhi, Salmonella typhimurium, Salmonella enteritidis, Salmonella mbandka, Salmonella bareilly, and Salmonella weltevreden, were detected, confirming this technique would be ideal for detection of the Salmonella serovars prevalent in seafood. This study also covered inhibition by the seafood matrix and the detection limit for dead Salmonella cells during the PCR assay. There was no visible inhibition of this Salmonella PCR assay by seafood matrices. The detection limit for dead Salmonella cells by 8-h PCR assay was 2 × 10³ CFU per 25 g seafood. The data indicated that dead cells of Salmonella in naturally contaminated seafood samples do not interfere with the assay resulting in false positives.