Association of urine glyphosate levels with renal injury biomarkers in children living close to major vegetable-producing regions in China.

Glyphosate (GLY)-based herbicides exposure contributes to renal dysfunction in experimental conditions, but the effects on humans are rarely reported. Biomonitoring is practically relevant for evaluating the association of urine GLY levels and renal damage in children living close to vegetable-cultivating regions. In this study, we collected the first-morning void urine samples of 239 healthy children (aged 3-12, 48.12 % boys) living near major vegetable-producing regions in March-May and August 2023 in Shandong Province, China. Urine levels of GLY and kidney injury-associated biomarkers were determined using ELISA kits to assess their correlation. GLY was detected in 92.05 % of urine samples (220 out of 239 participants) and the geometric concentration (GM) was 7.429 µg/L (range: 0.625 to 38.267 µg/L). Binary logistic regression and multivariate regression analysis revealed GLY detectability and levels positively correlated with home ventilation and self-producing vegetable intake of the subjects, as well as sampling periods. Moreover, a statistically significant concentration association with urine GLY was found for kidney injury-associated biomarkers (NGAL and KIM-1) (R2 = 0.923 and 0.855, respectively). Additionally, risk assessment revealed that the maximum value of probable daily intake was 0.150 mg/kg bw/day, accounting for 30.1 % of the established Acceptable Daily Intake of GLY. This study unveils a positive correlation between continuous GLY-based herbicide exposure and renal injury biomarkers of children. A large-scale epidemiological study is warranted for comprehensively assessing the effects of GLY-based herbicides on kidney function of the entire public.

Grape seed-derived procyanidin inhibits glyphosate-induced hepatocyte ferroptosis via enhancing crosstalk between Nrf2 and FGF12.

BACKGROUND: Glyphosate (GLY) exposure induces hepatocyte ferroptosis through overproduction of reactive oxygen species, regarded as an important contributor to liver damage. Grape seed-derived procyanidin (GSDP) has been reported to be an effective antioxidant, but whether and, if any, how GSDP can attenuate GLY-induced liver injury via inhibiting ferroptosis is unclear. PURPOSE: The current study aimed to investigate the hepato-protective effects and possible mechanisms of GSDP. METHODS: GLY-induced liver damage mice model was established to explore the hepatoprotective roles of GSPE in vivo. Subsequently, bioinformatics methodology was used to predict the key pathways and factors related to the action targets of GSPE against hepatocyte ferroptosis. Finally, we explored the roles of nuclear factor E2 related factor 2 (Nrf2) and fibroblast growth factor 21 (FGF21) in blunting GLY-induced liver damage via suppressing ferroptosis in vitro. RESULTS: GSDP exerts hepato-protective effects in vivo and in vitro through reduced oxidative stress and inhibited ferroptosis, which was related to the activation of Nrf2. Bioinformatics analysis showed an interaction between Nrf2 and FGF21. Furthermore, Nrf2 inhibition reduced FGF21 expression in the mRNA and protein levels. Fgf21 knockdown suppressed Nrf2 expression level, but recombinant FGF21 protein increased Nrf2 expression and promoted Nrf2 translocation into nucleus, suggesting a crosstalk between Nrf2 and FGF21. Intriguingly, the decreased levels of Nrf2 and FGF21 compromised the protective roles of GSDP against GLY-induced hepatocyte ferroptosis. CONCLUSION: These findings suggest that GSDP attenuates GLY-caused hepatocyte ferroptosis via enhancing the interplay between Nrf2 and FGF21. Thus, GSDP may be a promising natural compound to antagonize ferroptosis-related damage.

Measles epidemiology and viral nucleoprotein gene evolution in Shandong Province, China.

Measles, caused by measles virus (MeV), has not been eradicated in many regions and countries, threatening human health. Thus, it is beneficial for measles elimination to understand measles epidemiology and molecular evolution of key viral genes, such as nucleoprotein (N) gene. Based on public data, measles epidemiological information and MeV N gene sequences reported in Shandong Province, China were comprehensively collected and systematically analyzed. The results showed a positive correlation between population density and measles incidence (r = +0.31), while negative correlations were found between measles incidence and healthcare condition (r = -0.21) as well as average routine vaccination rate (r = -0.11). Additionally, the predominant lineage of MeV in Shandong was formed by genotype H1 strains, and the time of the most recent common ancestor of the N gene of MeV genotype H1 in Shandong traced back to 1987 (95% highest posterior density, 1984-1990) with relatively rapid evolution (mean rate, 1.267 × 10-3 substitutions/site/year). The genetic diversity of MeV N gene increased with the substantial emergence of major divergent clades of genotype H1 before 2005 and then remained relatively high and stable. In summary, these findings provided a significant insight into the measles elimination.

Prevalence and characterization of Clostridium perfringens isolated from different chicken farms in China.

Clostridium perfringens (C. perfringens) is a common pathogenic microorganism present in nature, which can cause animal and human diseases, such as necrotizing enteritis (NE) in poultry. Little is known about the current prevalence status of C. perfringens from poultry farms of different types and regions in China. From December 2018 to August 2019, we investigated the prevalence, genotype distribution and drug resistance of C. perfringens from Guangdong, Pingyin, Tai'an and Weifang. A total of 622 samples were collected and processed for C. perfringens isolation, among which 239 (38.42%) samples were determined to be positive for C. perfringens. A total of 312 isolates of C. perfringens were recovered (1-5 strains were isolated for each positive sample), and 98.72% of the isolates were identified as type A, while the others were type F. Antimicrobial susceptibility testing revealed that 47.71% of the isolates were resistant to at least five classes of commonly used antibiotics. Multilocus sequence typing (MLST) showed that 74 representative isolates were divided into 63 sequence types (STs), and the Simpson's diversity index (Ds) of the STs for the five farms was 0.9799. 37.84% of the isolates were classified into seven clonal complexes (CC1-CC7), and the isolates from the same farm were more concentrated in the minimum spanning tree. In addition, some cloaca isolates and feed isolates were distributed in the same ST or CC; this result indicates that the C. perfringens in chicken can come from the environment (feed etc.).

Prevalence and multilocus sequence typing of Clostridium perfringens isolated from retail chicken products and diseased chickens in Tai'an region, China.

BACKGROUND: Clostridium perfringens is an important zoonotic microorganism, which can cause diseases in animals and humans under suitable conditions. Contamination of C. perfringens in chicken products has been reported worldwide, but the genetic diversity and relationship of isolates were seldom analyzed. OBJECTIVES: The current study was undertaken to investigate the prevalence of C. perfringens from retail chicken products and sick chickens with suspected necrotic enteritis (NE) in Tai'an area, China. METHODS: In total, 295 samples were collected from Tai'an large poultry retail market and veterinary hospital in 2018, then the isolates were tested for toxin genes, drug resistance and multilocus sequence typing (MLST). RESULTS: Overall, 138 (46.78%) samples were determined to be positive for C. perfringens, and 99.37% of the isolates were identified as C. perfringens type A, with the remaining isolates being type F; 18.99% of the isolates were positive for cpb2 gene. Antimicrobial susceptibility testing revealed that 52.27% of the isolates from poultry retail market and diseased chickens showed multiple antibiotic resistance. MLST results showed that 50 analyzed isolates can be divided into 39 sequences types (STs), clustered in three clonal complexes (CCs) and 23 singletons. Although most of the isolates belong to type A, considerable genetic diversity can be observed, with the Simpson's diversity index up to 0.9181. MLST results and phylogenetic analysis showed that a portion of the isolates from humans and chickens were assigned to the same clusters in the phylogenetic tree or found to be in the same CCs, indicating the chicken isolates and the human isolates are related in certain stratification. CONCLUSIONS: This study showed that the contamination rate of C. perfringens in the local retail chicken products was relatively high. Most of the isolates exhibit broad-spectrum antimicrobial resistance. The high antibiotic resistance of C. perfringens isolates and the relationship between isolates from human and chicken indicated potential public health risks.

Farnesoid X receptor contributes to body weight-independent improvements in glycemic control after Roux-en-Y gastric bypass surgery in diet-induced obese mice.

OBJECTIVE: Roux-en-Y gastric bypass surgery (RYGB) can achieve long-term remission of type 2 diabetes. However, the specific molecular mechanism through which this occurs has remained largely elusive. Bile acid signaling through the nuclear hormone receptor farnesoid X receptor (FXR) exerts beneficial effects after sleeve gastrectomy (VSG), which has similar effects to RYGB. Therefore, we investigated whether FXR signaling is necessary to mediate glycemic control after RYGB. METHODS: RYGB or sham surgery was performed in high-fat diet-induced obese FXR-/- (knockout) and FXR+/+ (wild type) littermates. Sham-operated mice were fed ad libitum (S-AL) or by weight matching (S-WM) to RYGB mice via caloric restriction. Body weight, body composition, food intake, energy expenditure, glucose tolerance tests, insulin tolerance tests, and homeostatic model assessment of insulin resistance were performed. RESULTS: RYGB surgery decreases body weight and fat mass in WT and FXR-KO mice. RYGB surgery has similar effects on food intake and energy expenditure independent of genotype. In addition, body weight-independent improvements in glucose control were attenuated in FXR -/- relative to FXR +/+ mice after RYGB. Furthermore, pharmacologic blockade of the glucagon-like peptide-1 receptor (GLP-1R) blunts the glucoregulatory effects of RYGB in FXR +/+ but not in FXR -/- mice at 4 weeks after surgery. CONCLUSIONS: These results suggest that FXR signaling is not required for the weight loss up to 16 weeks after RYGB. Although most of the improvements in glucose homeostasis are secondary to RYGB-induced weight loss in wild type mice, FXR signaling contributes to glycemic control after RYGB in a body weight-independent manner, which might be mediated by an FXR-GLP-1 axis during the early postoperative period.

Occurrence and multilocus sequence typing of Clostridium perfringens isolated from retail duck products in Tai'an region, China.

Clostridium perfringens is an important zoonotic microorganism, which can cause diseases in animal and human under certain conditions. Contamination of C. perfringens in chicken and pork meat has been reported worldwide, but it is rarely reported in duck products. The current study was undertaken to investigate C. perfringens contamination in duck products from a large retail market in Tai'an region, China and the serotype distribution, antimicrobial resistance and genetic relatedness of the isolates. In total, 173 samples of duck products, 10 samples of environmental origins and 7 samples of fresh faeces from healthy shopkeepers were collected between March and November 2018, of which, 58 (31.69%), 10 (100%) and 7 (100%) samples were determined to be positive for C. perfringens, respectively. Ninety-nine isolates of C. perfringens were recovered, all of which were identified as type A. Beta2 (cpb2) toxin gene was found in 54.30% and 33.30% of the isolates from duck products and healthy shopkeepers, respectively. Antimicrobial susceptibility testing revealed that 90.10% of the isolates from duck products and environment showed multiple antibiotic resistance, among which, 49.40% were resistant to at least 6 classes of commonly used antibiotics. Multilocus sequence typing (MLST) showed that 58 representative isolates were divided into 41 sequences types (STs), among which, ST11 (8.60%) was the most common; 37.90% of all isolates were classified into four clonal complexes (CC1-CC4). The most prolific clonal complex (CC1), accounting for 24.13% of all isolates, contained isolates mainly from carcass, animal intestinal contents and environment of four retail stores. A portion of human isolates and duck isolates was distributed in the same CC or ST. In conclusion, C. perfringens contamination in some duck products in Tai'an retail market was relatively high, and most of the isolates exhibited broad-spectrum antimicrobial resistance. Although all the isolates belong to type A, considerable genetic diversity was observed, and a portion of the strains from human and duck was found to be phylogenetically close. The results indicated that antimicrobial-resistance strains of duck origin pose a potential threat to humans by spreading through the food chain.

Bacillus amyloliquefaciens BLCC1-0238 Can Effectively Improve Laying Performance and Egg Quality Via Enhancing Immunity and Regulating Reproductive Hormones of Laying Hens.

In this study, we sought to evaluate the effects of dietary Bacillus amyloliquefaciens (B. amyloliquefaciens) BLCC1-0238 supplementation on laying performance, egg quality, antioxidant enzyme activities, reproductive hormone, and immunity of laying hens. A total of 240 Hy-Line Brown laying hens (28 weeks old) were randomly divided into four groups, and three replicates per group (n = 20 per replicate). The control group was fed a standard basal diet, and the three treatment groups were provided the basal diet supplemented with either 0.01%, 0.03%, or 0.06% B. amyloliquefaciens BLCC1-0238 (2 × 1010 CFU/g), respectively. Hens were allowed 2 weeks to acclimate prior to initiation of the 8-week experiment. It was observed that dietary supplementation with 0.01% or 0.03% B. amyloliquefaciens BLCC1-0238 significantly increased egg production and egg mass. However, no significant differences in feed intake, egg weight, and feed conversion ratio among the four groups were observed. Different levels of B. amyloliquefaciens BLCC1-0238 supplementation also significantly increased egg shell strength and thickness. With respect to the levels of reproductive hormones in the hens, B. amyloliquefaciens BLCC1-0238 supplementation significantly reduced serum adrenal cortical hormone (ACTH) levels, while increasing estradiol (E2) and follicle-stimulating hormone (FSH) secretion in the treatment groups compared to the control group. Relative to the control group, supplementation with 0.03% and 0.06% B. amyloliquefaciens BLCC1-0238 was observed to significantly increase serum glutathione S-transferase (GST) concentration, and supplementation significantly reduced serum IL-1 and IL-6 levels, whereas IL-4 levels increased for all concentrations tested. In conclusion, supplementation of a basal chicken diet with B. amyloliquefaciens BLCC1-0238 can improve laying performance and egg quality through the reduction of stress responses, up-regulation of growth hormones, and supporting immunity in laying hens.

Prevalence and characterization of virulence genes in Escherichia coli isolated from piglets suffering post-weaning diarrhoea in Shandong Province, China.

The present study was performed to investigate the prevalence and characteristics of virulence genes in Escherichia coli (E. coli) isolated from piglets suffering post-weaning diarrhoea (PWD) in Shandong Province, China. The standard bacteriological method was used to isolate and identify E. coli, and then multiplex polymerase chain reaction (mPCR) was performed to determine virulence genes in E. coli. Among the 300 isolates, 166 (55.3％) harboured at least one virulence gene. Among the 166 isolates, 155 (93.4％) contained toxin-related genes. For enterotoxin genes, EAST1 (58/166, 34.9％) and LT-I (45/166, 27.1％) were the most common, followed by STa (32/166, 19.3％) and STb (21/166, 12.7％); for pathogenicity island (PAI) genes, irp2 (49/166, 29.5％) was the most dominant, followed by eae (48/166, 28.9％); for Shiga toxigenic E. coli (STEC)-associated toxin genes, Stx2e and hlyA genes were observed in 19 (19/166, 11.4％) and three strains (3/166, 1.8％) respectively. In addition, of the 166 isolates, 95 (95/166, 57.2％) contained adhesin genes, and AIDA-I (33/166, 19.9％) was the most common, followed by paa (27/166, 16.3％), F5 (K99) (20/166, 12.0％), F18 (15/166, 9.0％) and F41 (12/166, 7.2％). In summary, these findings demonstrated the prevalence and characteristics of virulence factors in E. coli isolates from piglets with PWD in Shandong Province of China, and the data may be useful for establishing preventive measures for post-weaning piglet diarrhoea.

A small-scale study on airborne transmission of H9N2 avian influenza virus under field conditions.

INTRODUCTION: H9N2 avian influenza viruses (AIV) can transmit in chicken flocks through direct contact and aerosols. Nevertheless, data on airborne transmission of AIV is very limited, especially under field conditions. To fill this literature gap, this study was designed to investigate airborne transmission of H9N2 AIV originating from infected chicken flocks under field conditions, with the aim to further characterize the airborne transmission of H9N2 AIV. METHODOLOGY: Oropharyngeal swabs were collected from different diseased chickens to confirm H9N2 AIV infection. All glass impingers 30 (AGI-30) were used to collect indoor, upwind and downwind air samples for three chicken houses with H9N2 AIV infected chickens. Swabs and air samples were tested for H9N2 AIV using a real-time reverse transcription polymerase chain reaction (RRT-PCR). H9N2 AIV was isolated in embryonated chicken eggs and hemagglutinin (HA) gene sequence similarity of the isolated AIV was compared. RESULTS: The results showed that indoor air samples were all RRT-PCR positive for H9N2 AIV. Downwind air samples collected between 10 m and 1.5 km away from the chicken houses were also found positive with an average load 2.62-5.21×103 RNA copies/m3. However, upwind air samples were all negative for H9N2 AIV. In addition, H9N2 AIV was isolated from swabs and indoor air samples. CONCLUSION: In summary, this study provides insights into the airborne transmission of H9N2 AIV under field conditions.

Prevalence and Antibiotic Resistance of Non-typhoidal Salmonella Isolated from Raw Chicken Carcasses of Commercial Broilers and Spent Hens in Tai'an, China.

The present study was aimed to determine the prevalence and characteristics of Salmonella isolated from meat samples of commercial broilers (CB) and spent hens (SH). Between March and June 2016, 200 retail raw chicken carcasses (100 from CB and 100 from SH) were obtained from local supermarkets in Tai'an city of China, and Salmonella isolates were then analyzed for antibiotic resistance, serotype, ß-lactamase genes, and the presence of class 1 integron. Forty Salmonella strains were obtained in this study (CB: 21/100, 21%; SH: 19/100, 19%). Three serotypes were identified in 40 Salmonella, and S. Enteritidis (CB: 15/21, 71.4%; SH: 10/19, 52.6%) was the dominant serotype, followed by S. Typhimurium (CB: 4/21, 19%; SH: 6/19, 31.6%) and S. Derby (CB: 2/21, 9.5%; SH: 3/19, 15.8%). Among 21 Salmonella isolated from CB, high antibiotic resistance rates were found for ampicillin (20/21, 95.2%), nalidixic acid (18/21, 85.7%), cefotaxime (17/21, 81%), and tetracycline (13/21, 61.9%); class 1 integron was observed in seven isolates (7/21, 33.3%), and gene cassettes included an empty integron (0.15 kb, n = 1), aadA2 (1.2 kb, n = 3), drfA1-aadA1 (1.4 kb, n = 1), and drfA17-aadA5 (1.7 kb, n = 2); blaTEM-1 was the dominant ß-lactamase gene (21/21, 100%), followed by blaCTX-M-55 (7/21, 33.3%). Among 19 Salmonella isolated from SH, high antibiotic resistance rates were found for nalidixic acid (19/19, 100%), tetracycline (19/19, 100%), ampicillin (18/19, 94.7%), and ciprofloxacin (13/19, 68.4%); class 1 integron was observed in two isolates (2/19, 10.5%), and gene cassettes included drfA17-aadA5 (1.7 kb, n = 1) and drfA1-aadA1 (1.4 kb, n = 1); blaTEM-1 was the dominant ß-lactamase gene (19/19, 100%), followed by blaCTX-M-55 (2/19, 10.5%) and blaCMY-2 (1/19, 5.3%). Collectively, antibiotic-resistant Salmonella can be widely detected in retail raw chicken carcasses of CB and SH, and therefore can pose a serious risk to public health.

Expression level of TNF-α in decidual tissue and peripheral blood of patients with recurrent spontaneous abortion.

OBJECTIVE: This study aimed to determine the express level of tumour necrosis factor α (TNF-α) in the decidual tissue and peripheral blood of patients with recurrent spontaneous abortion (RSA). MATERIAL AND METHODS: Eighty RSA patients and 100 control women were recruited in this study. Enzyme-linked immunosorbent assay (ELISA) was applied to determine the expression level of TNF-α in peripheral blood and decidual tissues from both groups. Additionally, the expression level of TNF-α was compared between RSA patients with different numbers of abortions, as well as primary and secondary RSA patients. RESULTS: The expression level of TNF-α in peripheral blood and decidual tissues of RSA patients was significantly higher compared to the controls (p < 0.001). Patients who had undergone RSA twice expressed TNF-α in peripheral blood and decidual tissues at a similar level to patients who had experienced RSA three times (p > 0.05), but significantly lower than patients who had experienced RSA more than three times (p < 0.001). The expression level of TNF-α in peripheral blood and decidual tissues was significantly higher in the secondary RSA patients, when compared with primary RSA patients (p < 0.001). CONCLUSIONS: Taken together, the relatively high expression level of TNF-α in decidual tissue and peripheral blood may be one of the causes of RSA and therefore could be used as a clinical indicator.

Prevalence and Antimicrobial Resistance of Salmonella Isolates Recovered from Retail Pork in Major Village Markets in Tai'an Region, China.

The current study was undertaken to evaluate Salmonella contamination in retail pork at major village markets of the Tai'an region, China. In total, 200 retail pork samples were collected from four village markets between June 2015 and February 2016, of which 69 samples (34.5%) were determined to be positive for Salmonella. Eleven serotypes were identified from the 69 Salmonella isolates, and Salmonella Derby was the most common (18 of 69, 26.1%), followed by Typhimurium (17 of 69, 24.6%) and Meleagridis (11 of 69, 15.9%). Antimicrobial susceptibility testing showed that antimicrobial resistance against tetracycline was the most prevalent (42 of 69, 60.9%), but antimicrobial resistance against both ceftriaxone and cefotaxime was 1.4% (1 of 69) and 2.9% (2 of 69), respectively. Multilocus sequence typing revealed that the 69 Salmonella isolates were divided into 11 sequence types (STs), among which ST40 (18 of 69, 26.1%) was the most common, followed by ST34 (15 of 69, 21.7%) and ST64 (13 of 69, 18.8%). Collectively, retail pork at village markets in the Tai'an region has a high Salmonella contamination rate, and these isolates exhibit broad-spectrum antimicrobial resistance. However, the absence of a dominant ST demonstrates that the Salmonella isolates from retail pork may be of diverse origins.

Characterization of ESBL-producing Escherichia coli recovered from companion dogs in Tai'an, China.

INTRODUCTION: Animals are considered to be reservoirs of extended-spectrum beta-lactamase (ESBL)-producing bacteria, but few epidemiological data on ESBL-producing Escherichia coli urinary tract isolates in pet dogs are available in China. METHODOLOGY: This study was conducted to describe the prevalence and characterization of ESBL producers among E. coli urinary tract isolates from pet dogs in Tai'an, China. RESULTS: A total of 118 E. coli were obtained from urinary samples of 80 companion dogs suffering from acute or chronic cystitis, of which three isolates from different dogs were ESBL producers. One isolate from dog A was of phylogroup A/ST410/CTX-M-15/TEM-1; one from dog B was of phylogroup B1/ST533/CTX-M-15/TEM-1; one from dog C was of phylogroup D/ST648/CTX-M-15. All ESBL producers were resistant to ampicillin, cephalexin, cefalotin, cefpodoxime, ceftiofur, enrofloxacin, marbofloxacin, and trimethoprim/sulfamethoxazole, but were susceptible to imipenem and amoxicillin/clavulanic acid. E. coli of ST533 carrying blaCTX-M-15 were first detected in pet dogs in China. CONCLUSIONS: Collectively, the findings could expand our knowledge about the prevalence and characterization of ESBL-producing E. coli urinary tract isolates in pet dogs in China.

Antimicrobial Resistance and Molecular Epidemiology of ESBL-Producing Escherichia coli Isolated from Outpatients in Town Hospitals of Shandong Province, China.

This study aimed to investigate antimicrobial resistance and molecular epidemiology of extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli (E. coli) isolated from outpatients in town hospitals of Shandong province, China. Antimicrobial susceptibility of ESBL-producing E. coli was tested using the disk diffusion and resistance genes encoding for ß-lactamases (blaTEM, blaCTXM, and blaSHV) were detected by polymerase chain reaction (PCR). Multilocus sequence typing (ST) of ESBL-producing E. coli was analyzed in this study. Our results showed that of 320 E. coli isolates, 201 carried ESBL genes (201/320, 62.8%), and these isolates all carried blaCTX-M genes, the most common being blaCTX-M-14 (116/201, 57.7%), followed by blaCTX-M-55 (47/201, 23.4%) and blaCTX-M-15 (31/201, 15.4%). ESBL-producing E. coli exhibited highly resistant to penicillin derivatives, fluoroquinolones, folate pathway inhibitors, and third-generation cephalosporins, but no carbapenem-resistant isolates were found in this study. Forty-two STs were found among the 201 ESBL-producing E. coli, and the most common ST was ST131 (27/201, 13.4%), followed by ST405 (19/201, 9.5%) and ST69 (15/201, 7.5%). Taken together, a high isolation rate of ESBL-producing E. coli (62.8%) was found among outpatients in town hospitals. blaCTX-M gene was most dominant and was composed of a variety of subtypes. No dominant ST was detected among ESBL-producing E. coli, indicating that these ESBL-producing E. coli isolates derive from different clones.

Detection and Characterization of Hepatitis E Virus in Goats at Slaughterhouse in Tai'an Region, China.

BACKGROUND: Hepatitis E virus (HEV) is a significant pathogen of viral hepatitis and can be transmitted through fecal-oral route. Epidemiological data concerning HEV in goats, however, are relatively sparse to date. Here, the prevalence and characteristics of HEV isolated from goats at slaughterhouse were investigated in Tai'an region, China. METHODS: Anti-HEV immunoglobulin G (IgG) in blood samples and HEV RNA in the liver samples were determined by using an enzyme-linked immunosorbent assay (ELISA) and a nested reverse transcription polymerase chain reaction (RT-PCR), respectively. In addition, partial nucleotide sequences of open reading frame 2 (ORF-2) of HEV isolates were analyzed. RESULTS: Fifty goat blood samples (46.7%, 50/120) were masculine for anti-HEV IgG. HEV RNA was detected in 2 liver samples (4.0%, 2/50) and belonged to genotype 4 subtype 4 h, with high identity (91.2-93%) with cow HEV strains detected in the same province, China. CONCLUSIONS: These findings demonstrated that goats may be an important reservoir for HEV and can become a major source of HEV infection in humans via food chain.

Prevalence and Antibiotic Resistance Profiles of Extended-Spectrum ß-Lactamase-Producing Escherichia coli Isolated from Healthy Broilers in Shandong Province, China.

Food-producing animals carrying extended-spectrum ß-lactamase-producing Escherichia coli (ESBL-EC) have posed a potential threat to human and animal health. However, information regarding ESBL-EC in the intensive broiler breeding areas of Shandong Province, People's Republic of China, is very limited. The goal of our study was to investigate the prevalence and drug resistance characteristics of ESBL-EC in healthy broilers from Shandong Province. A total of 142 ESBL-EC isolates were collected from four prefectures in Shandong Province from October 2014 to February 2015. ESBL-EC isolates were frequently detected (142 of 160 samples, 88.8%) in healthy broilers. Antibiotic susceptibility testing showed that all 142 ESBL-EC isolates were resistant to ampicillin, piperacillin, and cefazolin but were sensitive to imipenem and meropenem. All ESBL-EC isolates carried one or more of the bla genes, in which blaCTX-M, blaTEM-1, and blaSHV-5 genes were identified in 142, 106, and 5 isolates, respectively. The blaCTX-M gene includes blaCTX-M-15 (56), blaCTX-M-65 (42), blaCTX-M-55 (36), blaCTX-M-14 (21), blaCTX-M-79 (1), blaCTX-M-3 (1), blaCTX-M-123 (1), and blaCTX-M-132 (1). In addition, 17 ESBL-EC isolates cocarried the genes of the CTX-M-1 and CTX-M-9 groups. Our findings indicate that healthy broiler flocks in Shandong Province in China are an important reservoir for ESBL-EC, with blaCTX-M and blaTEM-1 being the prevalent resistance genes identified.

Avian influenza virus H9N2 seroprevalence and risk factors for infection in occupational poultry-exposed workers in Tai'an of China.

To determine risk factor for H9N2 avian influenza virus (AIV) infection, a serological surveillance among both occupational poultry-exposed (OPE) workers and general humans was carried out using both haemagglutination inhibition (HI) and microneutralization (MN) assays in Tai'an, China, between 2011 and 2013. At baseline, the positive rate of anti-H9 antibody (HI and MN titers ≥40) among OPE workers (51/600, 8.5%) was significantly higher than that among the general population (11/600, 1.8%). The result indicated that occupational exposure to chicken flocks was an important risk factor for H9N2 AIV infection. J. Med. Virol. 88:1453-1456, 2016. © 2016 Wiley Periodicals, Inc.

Phylogenetic Analysis of Hemagglutinin Genes of H9N2 Avian Influenza Viruses Isolated from Chickens in Shandong, China, between 1998 and 2013.

Since H9N2 avian influenza virus (AIV) was first isolated in Guangdong province of China, the virus has been circulating in chicken flocks in mainland China. However, a systematic phylogenetic analysis of H9N2 AIV from chickens in Shandong of China has not been conducted. Based on hemagglutinin (HA) gene sequences of H9N2 AIVs isolated from chickens in Shandong of China between 1998 and 2013, genetic evolution of 35 HA gene sequences was systematically analyzed in this study. Our findings showed that the majority of H9N2 AIVs (21 out of 35) belonged to the lineage h9.4.2.5. Most of isolates (33 out of 35) had a PSRSSR↓GLF motif in HA cleavage site. Importantly, 29 out of these 35 isolates had an amino acid exchange (Q226L) in the receptor-binding site. The substitution showed that H9N2 AIVs had the potential affinity to bind to human-like receptor. The currently prevalent H9N2 AIVs in Shandong belonged to the lineage h9.4.2.5 which are different from the vaccine strain SS/94 clade h9.4.2.3. Therefore, the long-term surveillance of H9N2 AIVs is of significance to combat the possible H9N2 AIV outbreaks.

Application of swine manure on agricultural fields contributes to extended-spectrum ß-lactamase-producing Escherichia coli spread in Tai'an, China.

The prevalence of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli (E. coli) is increasing rapidly in both hospital environments and animal farms. A lot of animal manure has been directly applied into arable fields in developing countries. But the impact of ESBL-positive bacteria from animal manure on the agricultural fields is sparse, especially in the rural regions of Tai'an, China. Here, we collected 29, 3, and 10 ESBL-producing E. coli from pig manure, compost, and soil samples, respectively. To track ESBL-harboring E. coli from agricultural soil, these isolates of different sources were analyzed with regard to antibiotic resistance profiles, ESBL genes, plasmid replicons, and enterobacterial repetitive intergenic consensus (ERIC)-polymerase chain reaction (PCR) typing. The results showed that all the isolates exhibited multi-drug resistant (MDR). CTX-M gene was the predominant ESBL gene in the isolates from pig farm samples (30/32, 93.8%) and soil samples (7/10, 70.0%), but no SHV gene was detected. Twenty-five isolates contained the IncF-type replicon of plasmid, including 18 strains (18/32, 56.3%) from the pig farm and 7 (7/10, 70.0%) from the soil samples. ERIC-PCR demonstrated that 3 isolates from soil had above 90% genetic similarity with strains from pig farm samples. In conclusion, application of animal manure carrying drug-resistant bacteria on agricultural fields is a likely contributor to antibiotic resistance gene spread.