Among-population variation in drought responses is consistent across life stages but not between native and non-native ranges.

Understanding how widespread species adapt to variation in abiotic conditions across their ranges is fundamental to ecology. Insight may come from studying how among-population variation (APV) in the common garden corresponds with the environmental conditions of source populations. However, there are no such studies comparing native vs non-native populations across multiple life stages. We examined APV in the performance and functional traits of 59 Conyza canadensis populations, in response to drought, across large aridity gradients in the native (North America) and non-native (Eurasia) ranges in three experiments. Our treatment (dry vs wet) was applied at the recruitment, juvenile, and adult life stages. We found contrasting patterns of APV in drought responses between the two ranges. In the native range, plant performance was less reduced by drought in populations from xeric than mesic habitats, but such relationship was not apparent for non-native populations. These range-specific patterns were consistent across the life stages. The weak adaptive responses of non-native populations indicate that they can become highly abundant even without complete local adaptation to abiotic environments and suggest that long-established invaders may still be evolving to the abiotic environment. These findings may explain lag times in invasions and raise concern about future expansions.

Occurrence, virulence, and resistance genes in Salmonella enterica isolated from an integrated poultry company in Jordan.

Salmonella is considered one of the most common foodborne pathogens worldwide. The annual number of hospitalizations and deaths related to zoonotic salmonellosis, which is transmitted from animals to humans and infects poultry and meat, is expected to be significant. Hence, the primary aims of this research were to isolate and characterize Salmonella species obtained from an integrated poultry company and identify some virulence, and antimicrobial resistance, with a specific concern about colistin resistance genes. A total of 635 samples collected from various sources in an integrated company in Jordan were screened for Salmonella species accompanying their virulence and antimicrobial resistance genes. Samples were collected from parent stock house drag swabs, broiler farms, premix, cecum at the slaughterhouse level, prechilling and postchilling stages, and the final product. Salmonella species were detected in 3% (6/200) of investigated parent stock house drag swabs, 13.8% (11/80) from cloacal swabs from broiler farms, 16.9% (11/65) from boiler farms premix, 24.4% (11/45) from the cecum at slaughterhouse level, 16.4% (9/55) from the prechilling stage, 37.8% (17/45) from the postchilling stage and 53.3% (24/45) from the final product stage. No isolates were detected in feed mills (0/20), parents' premix (0/40), or hatcheries (0/40). Salmonella isolates were resistant to ciprofloxacin (91.0%), nalidixic acid (86.5%), doxycycline (83.1%), tetracycline (83.1%), sulphamethoxazole-trimethoprim (79.8%) and ampicillin (76.4%). Serotyping shows that S. Infantis was the predominant serovar, with 56.2%. Based on the minimum inhibitory concentration (MIC) test, 39.3% (35/89) of the isolates were resistant to colistin; however, no mcr genes were detected. Among antimicrobial-resistant genes, blaTEM was the most prevalent (88.8%). Furthermore, the spvC, ompA, and ompF virulence genes showed the highest percentages (97.8%, 97.8%, and 96.6%, respectively). In conclusion, Salmonella isolates were found at various stages in the integrated company. S. Infantis was the most prevalent serotype. No mcr genes were detected. Cross-contamination between poultry production stages highlights the importance of good hygiene practices. Furthermore, the presence of virulence genes and the patterns of antimicrobial resistance present significant challenges for public health.

Phenotypic and genotypic characterization of Escherichia coli isolated from the chicken liver in relation to slaughterhouse conditions.

Avian pathogenic Escherichia coli (APEC) has been identified as a sub-group of extraintestinal pathogenic E. coli (ExPEC). Recent studies indicate APEC as a potential foodborne zoonotic pathogen and a source or reservoir of human extraintestinal infections. The slaughtering and processing of poultry in low-income countries such as Jordan occurs in two distinct ways: in informal facilities known as Natafat and in formal slaughterhouses. This study compared E. coli phenotypes and genotypes according to slaughtering conditions (formal slaughterhouses vs. informal slaughter facilities). Therefore, liver samples (n = 242) were collected from formal (n = 121) and informal slaughter facilities (n = 121). Results revealed a high prevalence (94.2%) of E. coli among all isolates, with 59 (17 formal and 42 informal) isolates considered avian pathogenic E. coli (APEC) based on the virulence-associated genes. The prevalence of resistance among isolates was relatively high, reaching up to 99% against penicillin and 97% against nalidixic acid. However, the prevalence of resistance was the lowest (1.3%) against both meropenem and imipenem. Based on the MIC test findings, colistin resistance was 46.9% (107/228). The mcr -1 gene prevalence was 51.4% (55/107), of which 17.1 % were from formal plants (6/36) and 68.1% from informal facilities (49/72). Interestingly, only one isolate (0.9%) expressed mcr-10. Escherichia coli O157:H7 and associated virulence genes were found more in informal (n = 15 genes) than in formal slaughterhouses (n = 8). Phylogroups B1, C, and A were the most frequent in 228 E. coli isolates, while G, B2, and clade were the least frequent. In conclusion, these findings highlight the importance of implementing biosecurity measures in slaughterhouses to reduce antibiotic-resistant E. coli spread. Furthermore, this study provides valuable insights into the effects of wet market (Natafat) slaughter conditions on increasing bacterial resistance and virulence.

Risk factors associated with mcr-1 colistin-resistance gene in Escherichia coli broiler samples in northern Jordan.

OBJECTIVES: The purpose of this study was to determine the prevalence of colistin-resistant Escherichia coli carrying mcr-1, and to identify risk factors associated with mcr gene-mediated resistance. METHODS: In total, 385 cloacal samples were collected from 125 broiler farms and a questionnaire containing information about each farm was designed and filled. RESULTS: Most of the antibiotics used in the disk diffusion method were highly resistant in all samples, with tetracycline and penicillin showing 100% and 99.7% resistance, respectively. Additionally, avian pathogenic E. coli (APEC) virulence genes frequency and percentage of APEC were identified, including sitA,iucC, and astA at 77%, 70.5%, and 62% respectively. In total, 214 of 360 isolates were positive for APEC (59.4%). Based on the minimum inhibitory (MIC) test, 58% of the isolates (n = 209 of 360) were resistant to colistin, with 39.7% displaying the mcr-1 gene. The statistical analysis of risk factors that influence colistin resistance prevalence revealed several significant factors, including commercial feed, farm management, sanitization, and antibiotic use. Irregular health checks for workers, non-dipping of feet before entering poultry houses, and the use of commercial poultry feeds all contributed to higher levels of colistin resistance as measured by MIC. On the other hand, doxycycline and commercial feed was 4 and 3.2 times more likely to occur based on the final logistic model of the mcr-1 gene, respectively. CONCLUSION: Our results suggest that better biosecurity protocols should be implemented in poultry farms to reduce antibiotic-resistant bacteria. Additionally, antibiotics should be carefully monitored and used only when necessary.

Multi-drug Resistant Acinetobacter baumannii: Phenotypic and Genotypic Resistance Profiles and the Associated Risk Factors in Teaching Hospital in Jordan.

BACKGROUND: This study aimed to determine the prevalence of Antimicrobial Resistance Genes (ARGs), with a focus on colistin resistance in clinical A. baumannii, as well as the risk factors associated with A. baumannii infection in Jordanian patients. METHODS: In total, 150 A. baumannii isolates were obtained from patients at a teaching hospital. The isolates were tested for antimicrobial susceptibility using disc diffusion and microdilution methods. PCR amplification was used to detect ARGs, and statistical analysis was conducted to evaluate the influence of identified risk factors on the ARGs acquisition. RESULTS: More than 90% of A. baumannii isolates were resistant to monobactam, carbapenem, cephalosporins, Fluoroquinolones, penicillin, and ß-lactam agents. Moreover, 20.6% of the isolates (n = 31) were colistin-resistant. Several ARGs were also detected in A. baumannii isolates. Univariate analysis indicated that risk factors and the carriage of ARGs were significantly associated P ≤ (0.05) with gender, invasive devices, immunodeficiency, systemic diseases, tumors, and covid-19. Logistic regression analysis indicated seven risk factors, and three protective factors were associated with the ARGs (armA, strA, and strB) P ≤ (0.05). In contrast, tetB and TEM were associated with 2 risk factors each P ≤ (0.05). CONCLUSION: Our study indicates a high prevalence of MDR A. baumannii infections in ICU patients, as well as describing the case of colistin-resistant A. baumannii for the first time in Jordan. Additionally, the risk factors associated with ARGs-producing A. baumannii infections among ICU patients suggest a rapid emergence and spread of MDR A. baumannii without adequate surveillance and control measures.

Mapping of infection prevention and control education and training in some countries of the World Health Organization's Eastern Mediterranean Region: current situation and future needs.

BACKGROUND: A strong understanding of infection prevention and control (IPC) procedures and comprehensive training among healthcare workers is essential for effective IPC programs. These elements play a crucial role in breaking the chain of nosocomial infections by preventing the transmission of resistant organisms to patients and staff members. This study mapped the components of IPC education and training across various member states of the World Health Organization (WHO) in the Eastern Mediterranean Region (EMR) at national, academic, and healthcare institutional levels. METHODS: A self-administered structured online questionnaire based on the WHO "Core Component 3" of IPC programs at the national and acute healthcare facility levels (IPC education and training) was given to national IPC focal persons in each of the WHO's EMR countries between February and March 2023. RESULTS: From 14 of the 22 countries,15 IPC persons participated in the survey. Most countries have scattered nonhomogeneous IPC education programs in human health undergraduate majors without considering it a standalone module. Academic institutions are rarely involved, and elaborate and predefined undergraduate IPC education programs provided by universities are present in 21.4% of the countries. In 71.4% of these countries, postgraduate training targeting IPC professionals is provided by national IPC teams, primarily based on national IPC guidelines developed with the aid of the WHO. Generally, healthcare worker training relies heavily on healthcare facilities in 92.9% of the countries, rather than on a national training program. In 42.9% of the countries, practicing IPC physicians are not necessarily specialists of infectious disease or medical microbiologists and IPC nurses are not required to specialize in IPC. However, nonspecialized IPC professionals are expected to undergo training upon employment and before beginning practice. Nongovernmental organizations such as the WHO play a significant role in IPC education and in supporting national IPC authorities in establishing national IPC guidelines, as it is the case in 78.6% of these countries. CONCLUSION: Clear disparities exist in IPC education and training across different countries in the WHO's EMR. Establishing a regional scientific network specializing in IPC would help bridge the existing gaps and standardize this education within individual countries and across countries in the region. This region needs to establish IPC certification standards and standardized education curricula.

First report of mobile colistin resistance gene mcr-1 in avian pathogenic Escherichia coli isolated from turkeys in the Gaza Strip, Palestine.

Background and Aim: Colistin is used to treat avian pathogenic Escherichia coli (APEC), a microorganism that affects turkey meat production in the Gaza Strip and worldwide. However, the recent emergence of plasmid-borne mobile colistin resistance (mcr) genes in pathogenic E. coli strains is a serious antimicrobial resistance (AMR) challenge for both human and animal health. In December 2018, colistin was banned as a veterinary antimicrobial in the Gaza Strip. This study aimed to detect and track the prevalence of colistin-resistant APEC isolated from turkey flocks in the Gaza Strip. Materials and Methods: This study investigated 239 APEC isolates from turkey flocks in the Gaza Strip between October 2018 and December 2021 (at 6-month intervals). The colistin-resistant APEC strains were detected using the broth microdilution method. The mcr-1 gene was identified using a polymerase chain reaction. Results: The overall colistin resistance among the isolated APECs was 32.2% during the study period. The average resistance in the first interval was 37.5%, which significantly decreased to 9.3% in the last interval. Among the 77 phenotypically resistant isolates, 32.4% were positive for mcr-1. The average abundance of mcr-1 in the first interval was 66.6%, which decreased to 25% in the last interval. Conclusion: To the best of our knowledge, this is the first study reporting the presence of the mcr-1 gene among the APEC isolates from turkeys in the Gaza Strip. Banned veterinary use of colistin significantly decreased the percentage of resistant APEC isolates from turkeys in Gaza Strip. Further studies are needed to investigate other colistin resistance genes and track the emergence of AMR.

Novel spa type and antimicrobial resistance gene profiles of Staphylococcus aureus strains isolated from bovine mastitis in Jordan.

Staphylococcu aureus is the most prevalent microorganism associated with mastitis in cattle. This study was designed to determine the spa types of Staph. aureus and to assess the resistance genes profile of isolated strains in dairy farms in Jordan. In total, 747 milk samples of cattle suffering from subclinical mastitis were collected from 37 dairy farms and tested for Staph. aureus. To detect antimicrobial resistance genes, all 219 strains of Staph. aureus were tested. Furthermore, 21 isolates of Staph. aureus were typed using spa typing. As a result, different proportions of resistance genes were found for Staph. aureus. High resistance genes were in tetK 100%, blaZ 99%, and tetM 97%. Moderate resistance genes were in aac(6')/aph(2'' 52%, ant(4')-Ia 48%, and ermC 41%. Low resistance genes were in ermA is 24%, aph(3')-III is 15%, and mecA is 15%. The spa typing of 21 isolates revealed six spa types, of which five were previously known. For the first time, a novel spa type (t17158) was identified as the main cause of mastitis in dairy cows in Jordan. The identification of resistance genes and spa types is helpful in determining the most effective treatments for cows and plays a significant role in reducing the transmission of pathogens.

Automated Monkeypox Skin Lesion Detection Using Deep Learning and Transfer Learning Techniques.

The current outbreak of monkeypox (mpox) has become a major public health concern because of the quick spread of this disease across multiple countries. Early detection and diagnosis of mpox is crucial for effective treatment and management. Considering this, the purpose of this research was to detect and validate the best performing model for detecting mpox using deep learning approaches and classification models. To achieve this goal, we evaluated the performance of five common pretrained deep learning models (VGG19, VGG16, ResNet50, MobileNetV2, and EfficientNetB3) and compared their accuracy levels when detecting mpox. The performance of the models was assessed with metrics (i.e., the accuracy, recall, precision, and F1-score). Our experimental results demonstrate that the MobileNetV2 model had the best classification performance with an accuracy level of 98.16%, a recall of 0.96, a precision of 0.99, and an F1-score of 0.98. Additionally, validation of the model with different datasets showed that the highest accuracy of 0.94% was achieved using the MobileNetV2 model. Our findings indicate that the MobileNetV2 method outperforms previous models described in the literature in mpox image classification. These results are promising, as they show that machine learning techniques could be used for the early detection of mpox. Our algorithm was able to achieve a high level of accuracy in classifying mpox in both the training and test sets, making it a potentially valuable tool for quick and accurate diagnosis in clinical settings.

Pathological and molecular study of kidneys in apparently healthy cattle and sheep with special reference to Leptospira species in central and northern Jordan.

Background and Aim: Renal pathological conditions can cause significant economic losses for livestock owners. This study investigated and described the gross pathology and histopathology of lesions found in the kidneys of sheep and cattle, with particular attention to the presence of Leptospira spp. Materials and Methods: Three hundred and sixty kidneys from apparently healthy sheep and cattle (208 and 152, respectively) were collected from slaughterhouses in Jordan for gross and histopathological examinations, multiplex polymerase chain reaction (PCR) analysis, and gene sequencing of Leptospira spp. Results: Histopathological analysis of the samples revealed the following conditions: interstitial nephritis (4.44%), glomerulonephritis (4.16%), acute tubular necrosis (17.22%), and renal congestion (4.72%). In addition, in 26.9% of the samples, hyaline material was observed in the interstitium of the cortex and medulla. The PCR results revealed that 83 (23.1%) of the 360 samples were positive for Leptospira spp. 16S rRNA, 42/152 (28%) of cattle, and 41/208 (20%) of sheep. Four samples (two sheep and two cattle) were sequenced and deposited in GenBank with accession numbers from OL701310 to OL701313. Basic Local Alignment Search Tool search and nucleotide similarities between OL701311 and OL701313 resulted in the highest similarities with different Leptospira borgpetersenii strains, whereas OL701310 showed the highest nucleotide similarity (99.2%) with the Leptospira interrogans strain. Similarly, phylogenetic analysis revealed that OL701311 to OL701313 clustered together with different serovars of L. borgpetersenii, whereas OL701310 clustered with the L. interrogans clade. Conclusion: This is the first study to reveal a close association between pathogenic Leptospira spp. and kidney disorders in Jordanian cattle and sheep. These findings may help expand the current understanding of the causes and mechanisms of renal disease in cattle and sheep and contribute to developing more effective prevention and treatment programs.

First molecular characterization of capsule expression and antibiotic susceptibility profile of Staphylococcus aureus isolates from bovine mastitis in Jordan.

Background and Aim: Bovine mastitis has long been considered the most important cause of economic losses in the dairy industry. Staphylococcus aureus is the most frequently isolated pathogen from bovine mastitis cases worldwide. Capsular polysaccharides (CPs) of serotype 5 (CP5) or serotype 8 (CP8) are the most prevalent capsule genotypes related to infections associated with S. aureus in humans. However, a variety of CPs has been reported in ruminants and other hosts. Information regarding the relationship between genotypic and phenotypic capsule variation and bovine mastitis in Jordan is scarce. Thus, we aimed to determine the prevalence of S. aureus capsule genotypes CP5 and CP8 in milk from bovine mastitis cases and the antimicrobial susceptibility profile of the recovered isolates in 27 dairy farms in Jordan. Materials and Methods: Staphylococcus aureus strains were isolated from bovine mastitis cases in two districts of Jordan. All S. aureus isolates were initially identified using conventional biochemical and microbiological methods. Subsequently, confirmation of the identity of S. aureus was performed by polymerase chain reaction (PCR) targeting nuc gene. Capsule polysaccharide typing was performed by PCR specific for CP5 and CP8. In addition, we assessed the antibiotic susceptibility profile of S. aureus isolates against commonly used antimicrobials by the disk diffusion method according to Clinical and Laboratory Standards Institute guidelines. Results: We collected 148 clinical isolates of S. aureus from bovine mastitis cases in the Zarqa (67.6%, n = 100) and Irbid (32.4%, n = 48) districts. Most isolates possessed capsule genotypes (91.3%), predominantly CP8 (88.6%). Only 8.7% of the isolates were nontypeable by PCR. In addition, we found statistically significant differences between the geographical region and the status of methicillin-resistant capsule genotypes (p < 0.05). The rates of resistance to ß-lactam, macrolide, and fluoroquinolone antibiotics were very low, but resistance to tetracyclines was considerably high (22.3%). Significantly, mastitis isolates from Irbid showed a higher rate of resistance to ciprofloxacin (8.3% vs. 0%), while isolates from Zarqa showed a significantly higher rate of resistance to gentamicin (12.0% vs. 6.2%). Conclusion: We established associations between capsule genotypes and antimicrobial resistance and the pathogenic behavior of S. aureus isolated from bovine mastitis cases. Further studies are necessary to fully elucidate the role and mechanisms of capsular expression in the epidemiological and molecular variability of S. aureus in bovine mastitis.

Homoisoflavonoids from the bulbs of Bellevalia longipes and an assessment of their potential cytotoxic activity.

Seven undescribed homoisoflavonoids were identified from the bulbs of Bellevalia longipes Post (Asparagaceae) as well as thirteen known and one natural homoisoflavonoid that had been reported as a synthetic product previously. A general approach for recognizing homoisoflavonoids via NMR spectroscopy data were presented. The undescribed compounds were: 8-dehydroxy-5-O-demethyl-6-hydroxyscillapersicone, 6-methoxyscillapersicone, 5-O-demethyl-6-methoxyscillapersicone, 8-O-methylscillapersicone, 4'-O-methylscillapersicone, 4',8-O,O-dimethylscillapersicone, 3'-O-methylscillapersicone, and 3-hydroxy-desmethylophiopogonanone A. Structures were determined based on analysis of HRMS and NMR data, while absolute configurations were assigned using ECD spectroscopy. Human cancer cell lines were used to assess the cytotoxic activities of the isolated compounds, where 3-dehydroxy-3'-hydroxyeucomol showed IC50 values of 0.62 µM, 5.36 µM, and 2.52 µM, when tested against MDA-MB-435 (melanoma), MDA-MB-231 (breast), and OVCAR3 (ovarian) cells, respectively.

Emergence of mcr-1 gene and carbapenemase-encoding genes among colistin-resistant Klebsiella pneumoniae clinical isolates in Jordan.

BACKGROUND: Antimicrobial resistance (AMR) is a global threat that requires serious attention, particularly when it is developed against colistin, which is considered one of the 'last-resort' antibiotics for curing an infection. This study aimed to investigate the AMR profile of the Klebsiella (K.) pneumoniae clinical isolates and to obtain the comprehensive characteristics of the carbapenemases among the carbapenem-resistant K. pneumoniae (CR-KP) when isolated. In addition, to detect the colistin resistance and investigate the MCR genes in the clinical K. pneumoniae isolates for the first time in Jordan. METHODS: A total of 179 K. pneumoniae isolates were cultured and they were confirmed using the VITEK 2 system and PCR. The antibiotic susceptibilities, extended-spectrum beta-lactamase (ESßL), multidrug-resistant (MDR), and CR-KP were determined by using the VITEK 2 system, disc diffusion, and the minimum inhibitory concentration (MIC) test. PCR was performed to detect the MCR and carbapenemase genes. RESULTS: The rates of ESßL, MDR, and CR-KP were 48 %, 62 %, and 12.8 %, respectively. High colistin resistance of 49.7 % (89/179) was found. Only one MCR-1 (1.1 %) out of the 89 colistin resistance isolates was detected. Many of the isolates harbored the ESßL genes. In particular, the carbapenem genes were detected in 26 isolates, with 46 % KPC enzyme genes (12/26), 23 % IMP genes (6/26), 19 % OXA-48 genes (5/26), 11.5 % NDM-1 genes (3/26) but no VIM gene was found. The statistical analyses revealed a significant association between colistin resistance and MDR (P ≤ 0.05, Chi-square test). An association between colistin resistance and the Piperacillin, Ceftazidime, Cefpodoxime, Imipenem, Aztreonam, and Tobramycin resistance was noted. CONCLUSION: The study's findings demonstrated the presence of the MCR-1 gene in the K. pneumoniae clinical isolates for the first time in Jordan and indicated that the KPC and IMP encoded carbapenemases were the most prevalent K. pneumoniae carbapenemases in Jordan patients.

Acquisition and evolution of enhanced mutualism-an underappreciated mechanism for invasive success?

Soil biota can determine plant invasiveness, yet biogeographical comparisons of microbial community composition and function across ranges are rare. We compared interactions between Conyza canadensis, a global plant invader, and arbuscular mycorrhizal (AM) fungi in 17 plant populations in each native and non-native range spanning similar climate and soil fertility gradients. We then grew seedlings in the greenhouse inoculated with AM fungi from the native range. In the field, Conyza plants were larger, more fecund, and associated with a richer community of more closely related AM fungal taxa in the non-native range. Fungal taxa that were more abundant in the non-native range also correlated positively with plant biomass, whereas taxa that were more abundant in the native range appeared parasitic. These patterns persisted when populations from both ranges were grown together in a greenhouse; non-native populations cultured a richer and more diverse AM fungal community and selected AM fungi that appeared to be more mutualistic. Our results provide experimental support for evolution toward enhanced mutualism in non-native ranges. Such novel relationships and the rapid evolution of mutualisms may contribute to the disproportionate abundance and impact of some non-native plant species.

UPLC-HRESI-MS and GC-MS analysis of the leaves of Nicotiana glauca.

The alkaloid-rich fraction obtained by fractionation of the crude methanolic extract of the leaves of wild tobacco tree Nicotiana glauca Graham (Solanaceae) was analyzed using UPLC-MS and GC-MS. Anabasine, a piperidine alkaloid, was identified as the major constituent with approximately 60 % (m/m) of the alkaloid-rich fraction. In addition to anabasine, six secondary metabolites were identified using high-resolution UPLC-MS. Anabasine was quantified in the leaves to be 1 mg g-1 dry plant material. The GC-MS analysis revealed five compounds with anabasine as the major component, while nicotine was not detected. Moreover, GC-MS was used for the analysis of the volatile oil that was obtained by hydro-distillation from the leaves of N. glauca. The volatile plant oil was found to be rich in oxygenated sesquiterpenes (e.g., ß-bisabolol) and carboxylic acids and esters (e.g., ethyl linoleate and hexadecanoic acid), whereas anabasine was not detected.

Assessment of Immune Response and Efficacy of Essential Oils Application on Controlling Necrotic Enteritis Induced by Clostridium perfringens in Broiler Chickens.

Necrotic enteritis (NE) caused by Clostridium perfringens is one of the most important enteric diseases in poultry. The antibacterial activity of two different essential oil (EO) blends against C. perfringens was investigated both in vitro and in vivo. Additionally, the immunological response to EO treatment was assessed. In the in vitro study, the antibacterial activity of EO formulas and commonly used antibiotics was evaluated against C. perfringens using disk diffusion assay, minimum inhibitory concentration (MIC) assay, and minimum bactericidal concentration (MBC) assay. In the in vivo study, NE experimental infection was performed on 440 Ross broiler chicks at 19 days of age for 4 continuous days. The chicks were treated with either EOs or amoxicillin at 22 days of age for 5 continuous days. One day after the end of treatment, the birds' performance was evaluated by calculating the feed conversion ratio. Serum samples from 120 birds were collected to measure the levels of IL-1ß, IFN-γ, IL-8, IL-10, and IL-17. After that, all birds were slaughtered, and their small intestines were subjected to gross and histopathological evaluation. In addition, bacterial counts in the small intestines were evaluated. In the in vitro study, EOs showed higher antimicrobial activities in comparison with antibiotics against C. perfringens. In the in vivo study, birds treated with EOs showed a significant decrease in bacterial counts, a significant decrease in intestinal lesions, and a significant improvement in performance compared with untreated birds (p < 0.05). Moreover, treating birds with EOs directed the immune system toward an anti-inflammatory pathway. None of the treated birds died due to NE compared with the 10% mortality rate in untreated birds. In conclusion, EOs might be an effective and safe alternative to antibiotics in the treatment of chicken NE.

Seroprevalence and risk factors of West Nile virus infection in veterinarians and horses in Northern Palestine.

BACKGROUND AND AIM: West Nile fever (WNF) is a neurotropic, mosquito-borne disease affecting humans and domesticated animals, caused by a member of the genus Flavivirus. Over the last decades, this virus has been responsible for several cases of illness in humans and animals. The current epidemiological status of WNF in horses is insufficient, and in veterinarians, as an occupational hazard is unknown. This study aimed to investigate and determine the seroprevalence and risk factors for WNF in veterinarians and horses in Palestine. MATERIALS AND METHODS: In this study, serum samples from 100 veterinarians and 87 horses were collected between August 2020 and September 2020 from different cities of Northern Palestine. West Nile virus (WNV) antibodies were detected using an enzyme-linked immunosorbent assay. RESULTS: Our results showed that 60.9% of the horse serum samples were positive in all investigated cities. In horses, location is a risk factor for the seropositivity for WNF, whereas age, sex, breed, and intended use of the horses, were not associated with increased WNF seropositivity. In veterinarians, 23.0% of the serum samples were positive. Positive samples were detected in all locations, age groups, experience length, and work sectors. However, the seropositivity for WNF was not influenced by these variables. CONCLUSION: The results revealed that WNV circulates in most regions of Palestine. Our results will help determine the risk of infection in animals and humans and control WNV transmission. Surveillance studies on humans, vectors, and animals are needed to better define endemic areas.

CCR2 Deficiency Impairs Ly6Clo and Ly6Chi Monocyte Responses in Orientia tsutsugamushi Infection.

Orientia (O.) tsutsugamushi, the causative agent of scrub typhus, is a neglected, obligate intracellular bacterium that has a prominent tropism for monocytes and macrophages. Complications often involve the lung, where interstitial pneumonia is a typical finding. The severity of scrub typhus in humans has been linked to altered plasma concentrations of chemokines which are known to act as chemoattractants for myeloid cells. The trafficking and function of monocyte responses is critically regulated by interaction of the CC chemokine ligand 2 (CCL2) and its CC chemokine receptor CCR2. In a self-healing mouse model of intradermal infection with the human-pathogenic Karp strain of O. tsutsugamushi, we investigated the role of CCR2 on bacterial dissemination, development of symptoms, lung histology and monocyte subsets in blood and lungs. CCR2-deficient mice showed a delayed onset of disease and resolution of symptoms, higher concentrations and impaired clearance of bacteria in the lung and the liver, accompanied by a slow infiltration of interstitial macrophages into the lungs. In the blood, we found an induction of circulating monocytes that depended on CCR2, while only a small increase in Ly6Chi monocytes was observed in CCR2-/- mice. In the lung, significantly higher numbers of Ly6Chi and Ly6Clo monocytes were found in the C57BL/6 mice compared to CCR2-/- mice. Both wildtype and CCR2-deficient mice developed an inflammatory milieu as shown by cytokine and inos/arg1 mRNA induction in the lung, but with delayed kinetics in CCR2-deficient mice. Histopathology revealed that infiltration of macrophages to the parenchyma, but not into the peribronchial tissue, depended on CCR2. In sum, our data suggest that in Orientia infection, CCR2 drives blood monocytosis and the influx and activation of Ly6Chi and Ly6Clo monocytes into the lung, thereby accelerating bacterial replication and development of interstitial pulmonary inflammation.

Phenethyisoquinoline alkaloids from the leaves of Androcymbium palaestinum.

Thirteen compounds were isolated from the methanolic extract of the leaves of Androcymbium palaestinum Baker (Colchicaceae). Of these, three were new, two were new natural products, and eight were known. The new isolated compounds were (+)-1-demethylandrocine (5), (-)-andropalaestine (8), and (+)-2-demethyl-ß-lumicolchicone (10), while the new natural products were (+)-O-methylkreysigine-N-oxide (3) and (+)-O,O-dimethylautumnaline (9). Moreover, two known compounds are reported for the first time from this species, specifically (-)-colchicine (11) and (-)-3-demethyldemecolcine (13). The structures of the isolated compounds were elucidated using a series of spectroscopic and spectrometric techniques, principally HRESIMS, 1D-NMR (1H and 13C NMR) and 2D-NMR (COSY, edited-HSQC, and HMBC). ECD spectroscopy was used for assigning the absolute configurations of compounds 3, 5, and 10. The cytotoxic activities of the isolated compounds were evaluated using the MDA-MB-435 (melanoma), MDA-MB-231 (breast), and OVCAR3 (ovary) cancer cell lines. Compound 11 was the most potent against all tested cell lines, with IC50 values of 12, 95 and 23 nM, respectively.

Identification of risk factors associated with antimicrobial resistance in equine fecal Escherichia coli isolates.

Antimicrobial resistance is a growing global problem that will need a multinational collaborative effort to overcome this serious challenge. The aim of the study is to investigate the potential risk factors associated with the prevalence and distribution of antimicrobial-resistance genes (ARGs) of Escherichia coli isolates obtained from equine fecal samples. One hundred and eighteen horses from different geographical locations and management systems were enrolled in the study and a questionnaire containing information about each individual horse was designed and filled. The enrolled horses belonged to 2 main categories (Hospitalized horses (n = 31), and Non hospitalized horses (n = 87)). In total, 103 E. coli isolates were collected from the 118 horse fecal samples. Genes that are responsible for resistance to ß-lactams, tetracyclines, aminoglycosides, and trimethoprim were detected using uniplex and multiplex polymerase chain reaction (PCR). The prevalence of antimicrobial resistance was significantly higher in hospitalized horses compared to non-hospitalized ones (p ≤ .05), particularly against (trimethoprim, doxycycline, oxytetracycline, and amoxicillin-clavulanic acid). The most prevalent antimicrobial-resistant genes were aminoglycoside resistant genes (strA, strB, and aadA) in percentages; 89%, 85%, and 84%, respectively. Statistical analysis revealed a significant association between risk factors and occurrence of ARGs (p ≤ .05). Significant risk factors include the last treatment and history of antimicrobial administration, breed of horses, use of horses, type of diet fed for horses, practice management and history of last illness. Tetracycline-resistance gene (tetA) was 23 times more likely to be found in the Arabian and local breeds of horses compared to English and warmblood breed. TetA is also 8 times more likely to be found in horses that were fed a natural diet compared to other horses that were fed manufactured/ processed feed. In conclusion, E. coli bacterium can harbor high resistance to different classes of antimicrobials which increases the risk of potential uncontrolled transmission of the multi-drug resistant E. coli bacterium to veterinarians and horse handlers, as well as to the equine population itself.