Escherichia coli and Salmonella enterica isolated from Egyptian dairy cattle herds: The prevalence and molecular characteristics.

Background: The pathogens Escherichia coli and Salmonella enterica that caused substantial health problems and financial losses were believed to have originated primarily from Egypt's dairy farms. Aim: The purpose of this study was to ascertain the occurrence of E. coli and S. enterica in three large dairy farms located in the Egyptian governorate of Sharkia. Furthermore, biochemical and serological characteristics of the isolated isolates were described. Further analysis revealed that several E. coli serovars had the genes stx1, stx2, eaeA, and hylA, while invA, stn, and hilA genes were found in several S. enterica serotypes using a multi-plex PCR. Methods: A total of 540 samples of fresh raw cow milk, water, feedstuffs, feces, (108 each), as well as swabs from feeders, milker hands and cattle crushes (36 each ), were gathered and analyzed. Results: The recovery of E. coli from various sampling sources was shown to have an overall prevalence of 62.2% (336/540) in the results. Fecal samples had isolated S. enterica, with a frequency of 0.74% (4/540). The existence of various groups of serovars, such as O26, O44, O55, O78 and O111 for E. coli and Salmonella enteritidis, Salmonella typhimurium and Salmonella inganda for S. enterica was revealed by serological identification of the two species. However, it was discovered that a number of E. coli serovars had much higher percentages of the eaeA and hylA genes as well as shiga-toxin types 1 and 2 (stx1 and stx2). The presence of the invA gene, a diagnostic marker for S. enterica was 100% across all serovars. Salmonella enteritidis possessed both the enterotoxin gene (stn) and the hyper-invasive locus gene (hilA). Salmonella typhimurium had the hilA gene, whereas S. inganda had the stn gene. Conclusion: Escherichia coli and S. enterica recovered in this study have significant genetic risk factors for high pathogenicity and virulence, posing a real threat to dairy population productivity and health, which could spread to the general public through milk.

Antibiotic Resistance, Virulence Gene Detection, and Biofilm Formation in Aeromonas spp. Isolated from Fish and Humans in Egypt.

The genus Aeromonas is widely distributed in aquatic environments and is recognized as a potential human pathogen. Some Aeromonas species are able to cause a wide spectrum of diseases, mainly gastroenteritis, skin and soft-tissue infections, bacteremia, and sepsis. The aim of the current study was to determine the prevalence of Aeromonas spp. in raw fish markets and humans in Zagazig, Egypt; identify the factors that contribute to virulence; determine the isolates' profile of antibiotic resistance; and to elucidate the ability of Aeromonas spp. to form biofilms. The examined samples included fish tissues and organs from tilapia (Oreochromis niloticus, n = 160) and mugil (Mugil cephalus, n = 105), and human skin swabs (n = 51) and fecal samples (n = 27). Based on biochemical and PCR assays, 11 isolates (3.2%) were confirmed as Aeromonas spp. and four isolates (1.2%) were confirmed as A. hydrophila. The virulence genes including haemolysin (hyl A) and aerolysin (aer) were detected using PCR in A. hydrophila in percentages of 25% and 50%, respectively. The antimicrobial resistance of Aeromonas spp. was assessed against 14 antibiotics comprising six classes. The resistance to cefixime (81.8%) and tobramycin (45.4%) was observed. The multiple antibiotic resistance (MAR) index ranged between 0.142-0.642 with 64.2% of the isolates having MAR values equal to 0.642. Biofilm formation capacity was assessed using a microtiter plate assay, and two isolates (18.1%) were classified as biofilm producers. This study establishes a baseline for monitoring and controlling the multidrug-resistant Aeromonas spp. and especially A. hydrophila in marine foods consumed in our country to protect humans and animals.

Detection and health risk assessment of toxic heavy metals in chilled and frozen meat collected from Sharkia province in Egypt.

Background: The consumption of meat is a fundamental aspect of global diets, providing essential nutrients and proteins vital for human nutrition. However, ensuring the safety of meat products has become progressively challenging due to potential contamination by toxic heavy metals (HMs) and pathogenic microorganisms. Aim: This study focuses on assessing the prevalence of Lead (Pb), Mercury (Hg), Arsenic (As), and Cadmium (Cd), in chilled and frozen meat in Sharkia Governorate, Egypt. Methods: A total of 30 samples, comprising 15 chilled and 15 frozen beef samples, were collected from various marketing stores in Sharkia. Analysis of toxic metals was conducted via atomic absorption spectrophotometer (AAS) following wet digestion. Results: The average levels (mg/kg) in chilled meat samples were found to be 0.64 ± 0.14 for Pb, undetectable for Hg, 0.02 ± 0.14 for Cd, and 4.66 ± 0.57 for As. In frozen samples, the average concentrations were 0.89 ± 0.21 for Pb, 0.08 ± 0.03 for Hg, 0.02 ± 0.004 Cd, and 5.32 ± 0.59 for As. Generally, the levels of HMs in frozen meat samples were observed to be higher than in chilled samples. Importantly, the levels of Pb were higher than maximum residual concentrations [maximum permissible limit (MPL)] in 53.3% of the chilled and 66.6% of the frozen, Cd levels in chilled and frozen were within the permissible concentrations in all samples, Hg was not identified in all the chilled and in 67% of frozen samples, and As levels were higher than the permissible levels in all samples chilled and frozen. The assessment of human health risk for adults revealed an estimated daily intake (EDI) value of beef meat below the threshold of the oral reference dose (RFD) for all analyzed metals except for As, where 46.7% of chilled samples and 60% of frozen samples exceeded the RFD. Furthermore, both the Hazard Quotient (THQ) for As and Hazard index (HI) for all the analyzed metals were above 1 in 33.3% of chilled samples and 46.7% of frozen samples. Conclusion: This indicates the remarkable adverse effects on human health associated with the consumption of meat with elevated levels of HMs, emphasizing the need for stringent quality control measures within the food industry.

Prevalence and bacterial isolation from hydatid cysts in dromedary camels (Camelus dromedarius) slaughtered at Sharkia abattoirs, Egypt.

Cystic echinococcosis (CE) is a severe neglected zoonotic parasitic disease caused by the larval stage of the dog tapeworm, Echinococcus granulosus. The objectives of this study were to determine the prevalence of hydatid cysts in dromedary camels (Camelus dromedarius) at Sharkia province, Egypt and investigate the occurrence of bacteria in hydatid fluid. A total of 6416 dromedary camels slaughtered in five abattoirs in Sharkia province, Egypt during the period from January and December 2018 were investigated for the presence of hydatid cysts. Furthermore, the bacterial species in 10 hydatid fluid isolated from lungs and livers was identified. The current findings revealed that the prevalence of hydatid cysts was 3.7%. Among those, the infection rate in lungs was 78.2%, which was significantly higher than hepatic infections (21.8%). The prevalence of hydatid cysts was the highest in winter (7.4%) and the lowest in spring (1.5%). The most common bacterial species found inside hydatid fluid collected from lungs were Salmonella spp., Staphylococcus spp., Enterococci and Pseudomonas spp. Meanwhile, Staphylococcus spp. were isolated from hepatic hydatid fluid. In conclusion, hydatid cysts infection is prevalent in dromedary camels in Sharkia province, Egypt as well as various aerobic and anaerobic bacterial species were isolated from hydatid fluid from camel lungs and livers.

Ringworm in calves: risk factors, improved molecular diagnosis, and therapeutic efficacy of an Aloe vera gel extract.

BACKGROUND: Dermatophytosis in calves is a major public and veterinary health concern worldwide because of its zoonotic potential and associated economic losses in cattle farms. However, this condition has lacked adequate attention; thus, to develop effective control measures, we determined ringworm prevalence, risk factors, and the direct-sample nested PCR diagnostic indices compared with the conventional methods of dermatophytes identification. Moreover, the phenolic composition of an Aloe vera gel extract (AGE) and its in vitro and in vivo antidermatophytic activity were evaluated and compared with those of antifungal drugs. RESULTS: Of the 760 calves examined, 55.79% (424/760) showed ringworm lesions; 84.91% (360/424) were positive for fungal elements in direct-microscopy, and 79.72% (338/424) were positive in culture. Trichophyton verrucosum was the most frequently identified dermatophyte (90.24%). The risk of dermatophytosis was higher in 4-6-month-old vs. 1-month-old calves (60% vs. 41%), and in summer and winter compared with spring and autumn seasons (66 and 54% vs. 48%). Poor hygienic conditions, intensive breeding systems, animal raising for meat production, parasitic infestation, crossbreeding, and newly purchased animals were statistically significant risk factors for dermatophytosis. One-step PCR targeting the conserved regions of the 18S and 28S genes achieved unequivocal identification of T. verrucosum and T. mentagrophytes in hair samples. Nested-PCR exhibited an excellent performance in all tested diagnostic indices and increased the species-specific detection of dermatophytes by 20% compared with culture. Terbinafine and miconazole were the most active antifungal agents for dermatophytes. Gallic acid, caffeic acid, chlorogenic acid, cinnamic acid, aloe-Emodin, quercetin, and rutin were the major phenolic compounds of AGE, as assessed using high-performance liquid chromatography (HPLC). These compounds increased and synergized the antidermatophytic activity of AGE. The treated groups showed significantly lower clinical scores vs. the control group (P < 0.05). The calves were successfully treated with topical AGE (500 ppm), resulting in clinical and mycological cure within 14-28 days of the experiment; however, the recovery was achieved earlier in the topical miconazole 2% and AGE plus oral terbinafine groups. CONCLUSIONS: The nested PCR assay provided a rapid diagnostic tool for dermatophytosis and complemented the conventional methods for initiating targeted treatments for ringworm in calves. The recognized antidermatophytic potential of AGE is an advantageous addition to the therapeutic outcomes of commercial drugs.

Monitoring of pesticides water pollution-The Egyptian River Nile.

BACKGROUND: Persistent organic pollutants represent about 95 % of the industrial sector effluents in Egypt. Contamination of the River Nile water with various pesticides poses a hazardous risk to both human and environmental compartments. Therefore, a large scale monitoring study was carried on pesticides pollution in three geographical main regions along the River Nil water stream, Egypt. METHODS: Organochlorine and organophosphorus pesticides were extracted by liquid-liquid extraction and analyzed by GC-ECD. RESULTS: Organochlorine pesticides mean concentrations along the River Nile water samples were 0.403, 1.081, 1.209, 3.22, and 1.192 µg L-1 for endrin, dieldrin, p, p'-DDD, p, p'-DDT, and p, p'-DDE, respectively. Dieldrin, p, p'-DDT, and p, p'-DDE were above the standard guidelines of the World Health Organization. Detected organophosphorus pesticides were Triazophos (2.601 µg L-1), Quinalphos (1.91 µg L-1), fenitrothion (1.222 µg L-1), Ethoprophos (1.076 µg L-1), chlorpyrifos (0.578 µg L-1), ethion (0.263 µg L-1), Fenamiphos (0.111 µg L-1), and pirimiphos-methyl (0.04 µg L-1). Toxicity characterization of organophosphorus pesticides according to water quality guidelines indicated the hazardous risk of detected chemicals to the public and to the different environmental compartments. The spatial distribution patterns of detected pesticides reflected the reverse relationship between regional temperature and organochlorine pesticides distribution. However, organophosphorus was distributed according to the local inputs of pollutant compounds. CONCLUSIONS: Toxicological and water quality standards data revealed the hazardous risk of detected pesticides in the Egyptian River Nile water to human and aquatic life. Thus, our monitoring data will provide viewpoints by which stricter legislation and regulatory controls can be admitted to avoid River Nile pesticide water pollution.

Listeria Species in Broiler Poultry Farms: Potential Public Health Hazards.

Broiler meat production worldwide has been plagued by lethal food-poisoning bacteria diseases, including listeriosis. A fatality rate of 15.6% was recorded in human beings in the EU in 2015. During 2013, a total of 200 poultry farm samples, including litter, chicken breast, farm feed, and drinking water, were collected to generate baseline data for the characterization of the genus Listeria in broiler poultry farms. Listeria spp. were detected in a total of 95 (47.5%) poultry farm samples. The isolates of Listeria spp. included L. innocua (28.5%), L. ivanovii (12.5%), L. welshimeri (4.5%), and L. monocytogenes and L. seeligeri (1% each). Listeria spp. contamination rates were higher in farm feed (70%), followed by litter (52.5%), chicken breasts (42.2%), and drinking water (10%). Almost all Listeria spp. isolates were resistant to more than three classes of antibiotics (multidrug resistant). Besides this, we observed a significant resistance level to penicillin and fluoroquinolone drugs. However, lower resistance levels were recorded for broad-spectrum cephalosporins. The inlA, inlC, and inlJ virulence genes were detected in almost all of the L. monocytogenes isolates. Thus, food safety management approaches and interventions at all stages of the broiler rearing cycle were needed to control cross-contamination and the zoonotic potential of listeriosis.

Veterinary antibiotic resistance, residues, and ecological risks in environmental samples obtained from poultry farms, Egypt.

In Egypt, poultry production constitutes one of the main sources of pollution with veterinary antibiotics (VAs) into the environment. About 80 % of meat production in Egypt is of poultry origin, and the potential environmental risks associated with the use of VAs in these farms have not yet been properly evaluated. Thus, the main purpose of this research was to evaluate the prevalence of antibiotic-resistant enteric key bacteria and the incidence of residual antibiotics in poultry farm environmental samples and to determine whether fertilizing soils with poultry litter from farms potentially brings ecological risks. From December 2011 to September 2012, a total of 225 litter, bird dropping, and water samples were collected from 75 randomly selected boiler poultry farms. A high prevalence of Escherichia coli (n = 179; 79.5 %) in contrast to the low prevalence of Salmonella spp. (n = 7; 3.1 %) was detected. Amongst E. coli isolates, serotypes O142:K86, O125:K70, O91:K, and O119:K69 were the most common. Meanwhile, Salmonella enterica serotypes emek and enteritidis were recovered. The antibiograms using the disc diffusion method revealed significantly more common resistant and multi-resistant isolates in broiler poultry farms. Residues of tetracycline and ciprofloxacin were detected at 2.125 and 1.401 mg kg(-1) mean levels, respectively, in environmental samples contaminated with E. coli-resistant strains by HPLC. The risk evaluations highlighted that tetracycline residues in poultry litter significantly display environmental risks with a hazard quotient value above 1 (1.64). Our study implies that ineffective implementation of veterinary laws which guide and guard against incorrect VA usage may potentially bring health and environmental risks.

Polychlorinated Biphenyls Water Pollution along the River Nile, Egypt.

Ten polychlorinated biphenyl (PCB) congeners were determined in water samples collected along the River Nile using gas chromatography-electron capture detector (GC-ECD). PCB concentrations ranged from 14 to 20 µg/L, which were higher than those reported in previous studies, indicating serious PCB pollution in the River Nile. PCB congener profiles varied depending on the sampling sties. PCB-138 was the predominant congener accounting for more than 18% of total PCBs. The composition of PCB congeners in the water revealed that highly chlorinated PCB technical mixtures such as Aroclor 1254 was the main PCB production historically used in Egypt. An increasing trend in PCB levels from the upper stream to the Nile estuaries was observed. The calculated flux of PCBs indicated that 6.8 tons of PCBs is dumped into the Mediterranean Sea each year from the River Nile. The hazard quotients and carcinogenic risk caused by PCB pollution in the River Nile were above the acceptable level indicating that PCBs in the River Nile water pose adverse health effects for all age groups. Our findings revealed that PCBs possess a serious risk to the Egyptian population that depends mainly on the River Nile as a source of water. Thus, stricter legislation and regulatory controls should be applied to reduce the risk of PCBs in Egypt.

Trace metal levels in water, fish, and sediment from River Nile, Egypt: potential health risks assessment.

The purposes of this study were to describe the impact of metal pollution on the main economic fish species Tilapia nilotica and to assess the potential health risk from consuming this contaminated fish in Egypt. Trace metals, including Ag, Al, Cd, Bo, Co, Cr, Cu, Fe, Mn, Mo, Ni, Pb, St, V, Zn, and As, were determined in water, Tilapia nilotica, and sediments from the River Nile, Domiate branch, Egypt. Metal concentrations in fish of Al, Cd, Co, Fe, Mn, Pb, V, and Zn (mg/kg dry weight [dw]) and concentrations in sediment of Cd, Co, Cr, Cu, Mn, Ni, Pb, V, and Zn (mg/kg dw) were above the International Atomic Energy Agency (IAEA-407) levels. However, trace metals in river water were still at permissible levels for Egyptian standards. The hazard index (HI) of estimated metal mixtures for intake of Tilapia nilotica (23.37) demonstrated that intake resulted in higher noncarcinogenic risk. In conclusion, the overall problem of metal contamination in fish collected from the River Nile was more serious than postulated to occur in an industrialized and densely populated area. In the light of known risks to public health, environmental protection laws are needed in Egypt.

Chronological Change of Resistance to ß-Lactams in Salmonella enterica serovar Infantis Isolated from Broilers in Japan.

Epidemiologic surveillance study was conducted in southern Japan to determine the antimicrobial resistance phenotypes and characterize the ß-lactamase genes and the plasmids harboring these genes in Salmonella enterica serovar Infantis (S. Infantis) isolates from broilers. Between January, 2007 and December, 2008, a total of 1,472 fecal samples were collected and examined at the Laboratory of Veterinary Public Health, Kagoshima University, Japan. In 93 (6.3%) isolates recovered, 33 (35.5%) isolates showed resistance to cefotaxime, an extended-spectrum cephalosporin (ESC), conferred by TEM-20, TEM-52 and CTX-M-25 extended-spectrum ß-lactamases (ESBLs). In addition to ESC-resistance, eight (8.6%) isolates exhibited resistance to cefoxitin mediated by CMY-2 AmpC ß-lactamase. Plasmid analysis and polymerase chain reaction replicon typing revealed the bla TEM-20 and bla CMY-2 genes were associated with IncP plasmids, bla TEM-52 was linked with a non-typable plasmid and bla CTX-M-25 was carried by an IncA/C plasmid. Non-ß-lactam resistance to streptomycin, sulfamethoxazole, and oxytetracycline encoded by the aadA1, sul1, and tet(A) genes, respectively, was found in 86 (92.5%) isolates. Resistance to kanamycin and ofloxacin was exhibited in 12 (12.9%) and 11 (11.8%) isolates, respectively, the former was mediated by aphA1-Iab. These data indicate that S. Infantis isolates producing ESBLs and AmpC ß-lactamase have spread among broiler farms in Japan. These data demonstrated that the incidence of ESC-resistant S. Infantis carrying bla TEM-52 remarkably increased and S. Infantis strains harboring bla CMY-2, bla TEM-20, or bla CTX-M-25 genes emerged from broilers in Japan for the first time in 2007 and 2008.

Re-emergence of multi-drug resistant Salmonella enterica serovar Stanley from cattle.

During 2009, Salmonella enterica subspecies enterica serovar Stanley isolates were recovered from cattle diagnostic specimens in southern Japan, and the isolates were examined to characterize the genetic determinants involved in this new pathogenicity that associated with mortality in cattle. All the isolates were multi-drug resistance exhibited resistance to ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, oxytetracycline, and kanamycin (ACSSuT-Km) encoded by blaTEM, catA, aadA1, sul1, tet(A), and aphA1 genes, respectively. Class 1 integrons of 1.5-kb size were detected in all MDR isolates. The isolates harboured easily transferable plasmids of ca. 210-kb with the potential of transmitting resistance phenotype and genotype detected in the donor isolates. XbaI-digested PFGE patterns generated two related clusters implicated in the dissemination of multi-drug resistance amongst Salmonella Stanley isolates. An emergence of multi-drug resistant Salmonella Stanley amongst food-producing animals, including cattle is a threat to human health, as resistant isolates may be transmitted to humans through the food chain.

Characterization of antibiotic resistance and the emergence of AmpC-producing Salmonella Infantis from pigs.

During the period of 2007-2008, a total of 270 pig fecal samples were collected from a meat processing plant located in southern Japan and examined for Salmonella species. A total of 44 Salmonella isolates were recovered, and antimicrobial resistance was detected in serotypes Typhimurium (n=9), Infantis and Choleraesuis (n=2), and Derby, Miyazaki and Schwarzengrund (n=1). Multidrug resistance was seen in serotypes Typhimurium (n=8) and Infantis (n=2). The most commonly observed resistance phenotypes were against streptomycin, oxytetracycline and sulfamethoxazole (100%), ampicillin (90%), chloramphenicol (50%), cephalothin (30%) and cefoxitin, ceftazidime and kanamycin (each 20%). Extended-spectrum cephalosporin-resistant Salmonella Infantis isolates producing plasmid-mediated, bla(CMY-2) gene were detected. These AmpC-producing isolates showed resistance to ampicillin and cephems (cephalothin, cefoxitin and ceftazidime). Resistance transfer experiments showed that transconjugants and transformants coexpressed resistance phenotypes similar to the donor isolates. To the best of our knowledge, this is the first report worldwide describing serovar Infantis from pigs capable of producing AmpC ß-lactamase. Then, we detected the pentadrug-resistance phenotype in Salmonella Typhimurium isolates, which yielded class 1 integron amplicons of 1.0 and 1.2 kb. Genetic fingerprinting analysis by pulsed-field gel electrophoresis and an assay by polymerase chain reaction confirmed the isolates to be Salmonella Typhimurium DT104. In conclusion, the findings of this survey call for the systematic and comprehensive domestic and international surveillance programs to determine the true rates of occurrence of AmpC-producing Salmonella both in the livestock and public health sectors.

Genetic analysis of multidrug-resistant Salmonella enterica serovars Stanley and Typhimurium from cattle.

During 2005-2008, a longitudinal study was conducted in southern Japan to detect and characterize multidrug-resistant Salmonella enterica serovars recovered from cattle diagnostic specimens. Determination of antimicrobial resistance phenotypes and genotypes, identification of Salmonella genomic island 1 (SGI1), detection of virulence genes, plasmid analysis, conjugal transfer experiments, and sequencing of class 1 integrons were conducted. Multidrug-resistant Salmonella detected were serovars Stanley, Typhimurium, and O4:d. Salmonella Stanley isolates exhibited resistance to ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, oxytetracycline, trimethoprim, and kanamycin (ACSSuT+) encoded by bla(TEM), catA, aadA2, tetA, sul1, dfrA12, and aphA1 genes, respectively. Sequencing analysis revealed that aadA2 and dfrA12 were integrated as gene cassettes within the class 1 integrons of 1.5kb size. Importantly, the isolates harboured easily transferable plasmids of ca. 210kb with the potential of transmitting resistance phenotype and genotype detected in the donor isolates. Moreover, Salmonella Typhimurium DT104 isolates with typical SGI1 were detected and presented ACSSuT+ resistance phenotype encoded by bla(PSE-1) and bla(TEM); floR; aadA1; sul1; and tetA and tetG, respectively. Salmonella Typhimurium isolates carried plasmids of variable sizes ranging from 3.5 to 100 kb with DT104 isolates harbouring plasmids of ca. 90 kb. Salmonella serovar O4:d had ACSSuT+ resistance phenotype mediated by bla(TEM), catA, aadA1, sul1, tetA, and aphA1 genes. A virulence gene invA was found in all multidrug-resistant Salmonella Typhimurium, Stanley and O4:d clinical isolates. In conclusion, this is the first report describing the occurrence of multidrug-resistant Salmonella Stanley from bovine species. The emergence of Salmonella Stanley isolates exhibiting plasmid-encoded high-level multidrug resistance is an important health concern because this new pathogenecity was associated with mortality in cattle.

Detection and characterization of extended-spectrum beta-lactamase (TEM-52)-producing Salmonella serotype Infantis from broilers in Japan.

During 2004 and 2006, multidrug-resistant Salmonella enterica subspecies enterica serovar Infantis (Salmonella Infantis) isolates (n = 120) were recovered from broiler cecal samples collected from a meat-processing plant, and the isolates were examined. The study was conducted to detect and characterize extended-spectrum beta-lactamase (ESBL)-producing Salmonella Infantis isolates recovered from broiler chickens and determine the mechanisms of transfer of the resistance traits. Extended-spectrum cephalosporins-resistant Salmonella Infantis isolates producing ESBL TEM-52 were detected. The mutant bla(TEM-52) gene and the wild-type bla(TEM-1) gene that mediated resistance to ampicillin (an extended-spectrum penicillin) and cephalothin (a narrow-spectrum cephalosporin) were located on approximately 50-kb conjugative plasmids among beta-lactam-resistant (n = 29) isolates. The bla(TEM) genes did not cotransfer with aadA1, sul1 (both associated with class 1 integrons), tetA, and dfrA5, signifying a chromosomal location of these non-beta-lactam resistance-encoding genes. This is the first report describing TEM-52-producing S. enterica from food-producing animals in Japan. An emergence of TEM-type ESBL is an important concern to public health because this readily transferable resistance mechanism threatens the value of the third-generation cephalosporins and may reduce the clinical utility of this class of antibiotics against pathogenic Gram-negative bacteria.