Publications by authors named "Heinrich Neubauer"

197 Publications

Serosurvey and Risk Factors Associated with Infection in High Risk Occupations from District Lahore and Kasur of Punjab, Pakistan.

Pathogens 2021 May 18;10(5). Epub 2021 May 18.

Institute of Bacterial Infections and Zoonoses, Friedrich-Loeffler-Institut, Naumburger Str. 96a, 07743 Jena, Germany.

Brucellosis is a neglected zoonotic disease, and occupations with a high risk of infection exist. Limited information is available on brucellosis for these employees at high risk in Pakistan. A total of 459 persons with high-risk occupations, i.e., 211 abattoir workers, 63 milkers, 52 dung cake makers, 44 veterinarians, 44 shepherds, and 45 veterinary students of the districts Kasur and Lahore, Pakistan, were tested in this study. Blood samples and information on place of residence, gender, age, urbanicity, type of occupation, socioeconomic status, contact with animals, consumption of raw milk, contact to women who had a miscarriage, contact to aborted animal fetus or abortion material, pregnancy, miscarriage history, or intrauterine fetal death were collected. Serum samples were examined using Rose Bengal tests for anti- antibodies and seropositive samples were subjected to genus-specific qPCR for the detection of DNA. Data were analyzed using chi-squared and binary regression. Twenty (4.35%) persons were seropositive for anti- antibodies. Out of these, 18 (90%) were tested positive by genus-specific qPCR. Positive sera were more often found in Lahore district (8.3%) than in Kasur district (3.1%). Persons older than 36 years were more often seropositive. Persons involved in the handing of milk and in contact with animals were more often seropositive. Contact with women who had had a miscarriage was also a significant factor for seropositivity for brucellosis. Place of residence, gender, age, urbanicity, and contact with animals were identified as potential risk factors in the present study. The present study confirmed that brucellosis is an occupation hazard for abattoir workers, milkers, dung cake makers, and veterinarians in the districts of Lahore and Kasur of Punjab, Pakistan. The study shows the need for strategies for safety at work to minimize the risk of infection. Raising awareness for the prevention and use of proper personal protection equipment during the slaughtering and treatment of animal is highly needed.
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http://dx.doi.org/10.3390/pathogens10050620DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8158371PMC
May 2021

WGS-Based Analysis of Carbapenem-Resistant in Vietnam and Molecular Characterization of Antimicrobial Determinants and MLST in Southeast Asia.

Antibiotics (Basel) 2021 May 11;10(5). Epub 2021 May 11.

Friedrich-Loeffler-Institute, Institute of Bacterial Infections and Zoonoses (IBIZ), Naumburger str.96a, D-07743 Jena, Germany.

Carbapenem-resistant (, CRAb) is an emerging global threat for healthcare systems, particularly in Southeast Asia. Next-generation sequencing (NGS) technology was employed to map genes associated with antimicrobial resistance (AMR) and to identify multilocus sequence types (MLST). Eleven strains isolated from humans in Vietnam were sequenced, and their AMR genes and MLST were compared to published genomes of strains originating from Southeast Asia, i.e., Thailand ( = 49), Myanmar ( = 38), Malaysia ( = 11), Singapore ( = 4) and Taiwan ( = 1). Ten out of eleven Vietnamese strains were CRAb and were susceptible only to colistin. All strains harbored (3")-IIa, A, (6)-Id and (3") genes conferring resistance to aminoglycosides, and OXA-51 variants and ADC-25 conferring resistance to ß-lactams. More than half of the strains harbored genes that confer resistance to tetracyclines, sulfonamides and macrolides. The strains showed high diversity, where six were assigned to sequence type (ST)/2, and two were allocated to two new STs (ST/1411-1412). MLST analyses of 108 strains from Southeast Asia identified 19 sequence types (ST), and ST/2 was the most prevalent found in 62 strains. A broad range of AMR genes was identified mediating resistance to ß-lactams, including cephalosporins and carbapenems (e.g., OXA-51-like, OXA-23, ADC-25, ADC-73, TEM-1, NDM-1), aminoglycosides (e.g., (3")-IIa, (3")-Ib, (6)-Id, A and (3')-Ia), phenicoles (e.g., B8), tetracyclines (e.g., B and .39), sulfonamides (e.g., .1 and sul.2), macrolides and lincosamide (e.g., .E, .E and F). MLST and core genome MLST (cgMLST) showed an extreme diversity among the strains. Several strains isolated from different countries clustered together by cgMLST; however, different clusters shared the same ST. Developing an action plan on AMR, increasing awareness and prohibiting the selling of antibiotics without prescription must be mandatory for this region. Such efforts are critical for enforcing targeted policies on the rational use of carbapenem compounds and controlling AMR dissemination and emergence in general.
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http://dx.doi.org/10.3390/antibiotics10050563DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8150915PMC
May 2021

Molecular characterization and antimicrobial susceptibility testing of clinical and non-clinical and isolates from Egypt.

One Health 2021 Dec 27;13:100255. Epub 2021 Apr 27.

Friedrich-Loeffler-Institut, Institute of Bacterial Infections and Zoonoses, Naumburger Str. 96a, 07743 Jena, Germany.

Brucellosis is a highly contagious and incapacitating disease of humans, livestock and wildlife species globally. Treatment of brucellosis in animals is not recommended, and in humans, combinations of antibiotics recommended by the World Health Organization are used. However, sporadic antimicrobial-resistant (AMR) isolates and relapse cases have been reported from different endemic regions. In the current study, molecular characterization and antibiotic susceptibility testing using the microdilution method for 35 and strains isolated from humans, milk and animal were carried out. Additionally, Next-Generation-Sequencing (NGS) technology was applied to confirm at the species level and investigate AMR and pathogenicity-associated determinants. MALDI-TOF seemed to be a rapid and reliable tool for routine identification of brucellae to the genus level; however, DNA-based identification is indispensable for accurate species identification. strains were isolated from two human cases and a sheep. Such infections are uncommon in Egypt. Egyptian strains are still in-vitro susceptible to doxycycline, tetracyclines, gentamicin, ciprofloxacin, levofloxacin, chloramphenicol, streptomycin, trimethoprim/sulfamethoxazole and tigecycline. Probable (no CLSI/EUCAST breakpoints have been defined yet) in-vitro resistance to rifampicin and azithromycin was observed. WGS failed to determine classical AMR genes, and no difference in the distribution of virulence-associated genes in all isolates was found. Isolates of human and non-human origins were still susceptible to the majority of antibiotics used for treatment in humans. The absence of classical AMR genes in genomes of "resistant" strains may reflect a lack of information in databases, or resistance might not be encoded by single resistance genes. The One Health approach is necessary for tackling brucellosis. Continuous susceptibility testing, updating of breakpoints, assessing mutations that lead to resistance are needed.
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http://dx.doi.org/10.1016/j.onehlt.2021.100255DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8122161PMC
December 2021

Molecular Typing of Burkholderia mallei Isolates from Equids with Glanders, India.

Emerg Infect Dis 2021 Jun;27(6):1745-1748

We collected 10 Burkholderia mallei isolates from equids in 9 districts in India during glanders outbreaks in 2013-2016. Multilocus variable-number tandem-repeat analysis showed 7 outbreak area-related genotypes. The study highlights the utility of this analysis for epidemiologically tracing of specific B. mallei isolates during outbreaks.
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http://dx.doi.org/10.3201/eid2706.203232DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8153868PMC
June 2021

No hints at glyphosate-induced ruminal dysbiosis in cows.

NPJ Biofilms Microbiomes 2021 03 25;7(1):30. Epub 2021 Mar 25.

Institute of Animal Nutrition, Federal Research Institute for Animal Health, Friedrich-Loeffler-Institute, Brunswick, Germany.

Glyphosate-based herbicides are among the most used non-selective herbicides worldwide and inhibit synthesis of aromatic amino acids in plants, bacteria, and fungi. Given the broad usage, controversies concerning potential effects of glyphosate on health and especially on gut microbiomes arose. For cattle, it has been proposed based on in vitro data that glyphosate has detrimental effects on the ruminal microbiome, which manifest as a specific inhibition of bacteria involved in fiber degradation and as an enrichment of specific pathogens. In the present study, glyphosate effects on the ruminal microbiome were analyzed in vivo using glyphosate contaminated feedstuffs with strong differences in dietary fiber and dietary energy content in order to reproduce the proposed detrimental glyphosate effects on the rumen microbiome. While significant impact of dietary factors on the ruminal microbiome and its products are pointed out, no adverse glyphosate effects on ruminal microbiome composition, diversity, and microbial metabolites are observed.
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http://dx.doi.org/10.1038/s41522-021-00198-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7994389PMC
March 2021

Comparative in silico genome analysis of Clostridium perfringens unravels stable phylogroups with different genome characteristics and pathogenic potential.

Sci Rep 2021 Mar 24;11(1):6756. Epub 2021 Mar 24.

Institute of Bacterial Infections and Zoonoses, Friedrich-Loeffler-Institut, Naumburger Str. 96A, 07743, Jena, Germany.

Clostridium perfringens causes a plethora of devastating infections, with toxin production being the underlying mechanism of pathogenicity in various hosts. Genomic analyses of 206 public-available C. perfringens strains´ sequence data identified a substantial degree of genomic variability in respect to episome content, chromosome size and mobile elements. However, the position and order of the local collinear blocks on the chromosome showed a considerable degree of preservation. The strains were divided into five stable phylogroups (I-V). Phylogroup I contained human food poisoning strains with chromosomal enterotoxin (cpe) and a Darmbrand strain characterized by a high frequency of mobile elements, a relatively small genome size and a marked loss of chromosomal genes, including loss of genes encoding virulence traits. These features might correspond to the adaptation of these strains to a particular habitat, causing human foodborne illnesses. This contrasts strains that belong to phylogroup II where the genome size points to the acquisition of genetic material. Most strains of phylogroup II have been isolated from enteric lesions in horses and dogs. Phylogroups III, IV and V are heterogeneous groups containing a variety of different strains, with phylogroup III being the most abundant (65.5%). In conclusion, C. perfringens displays five stable phylogroups reflecting different disease involvements, prompting further studies on the evolution of this highly important pathogen.
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http://dx.doi.org/10.1038/s41598-021-86148-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7991664PMC
March 2021

Validation of a Commercial Glanders ELISA as an Alternative to the CFT in International Trade of Equidae.

Front Vet Sci 2021 16;8:628389. Epub 2021 Feb 16.

Friedrich-Loeffler-Institut (FLI), Federal Research Institute for Animal Health, Institute of Bacterial Infections and Zoonoses, Jena, Germany.

Glanders, caused by (.) is a notifiable zoonotic disease in equidae. For international trade and movement of equids, certificates of negative serological test results for antibodies against are required. To date, the complement fixation test (CFT) is the mandatory test to issue these health certificates. The CFT is difficult to standardize and, due to its poor specificity, often leads to false-positive reactions resulting in trade restrictions with considerable financial consequences. In the present study, the new ID Screen Glanders Double Antigen Multispecies ELISA (GLANDA- ELISA) (IDvet, Grabels, France) was evaluated using 400 negative and 370 glanders positive field samples of equidae. The GLANDA-ELISA was significantly more specific (99.8%) than the CFT (97.0%). Considering the comparable sensitivities of CFT (96.5%) and ELISA (98.1%), this new GLANDA-ELISA test appears a suitable confirmatory test and a realistic alternative for serological testing of horses for trade or movement.
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http://dx.doi.org/10.3389/fvets.2021.628389DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7920956PMC
February 2021

Identification, geographic distribution and risk factors of Brucella abortus and Brucella melitensis infection in cattle in Algeria.

Vet Microbiol 2021 Mar 29;254:109004. Epub 2021 Jan 29.

Friedrich-Loeffler-Institut, Institute of Bacterial Infections and Zoonoses, 07743, Jena, Germany; Faculty of Veterinary Medicine, Kafrelsheikh University, 33516, Kafr El-Sheikh, Egypt. Electronic address:

Brucellosis is an infectious disease of several terrestrial and marine animals and humans caused by bacteria of the genus Brucella. This study aimed to identify Brucella species and biovars circulating in cattle and to analyze their geographic distribution across Algeria. Two hundred ninety eight milk and lymph node samples from 161 seropositive cattle of different local and foreign breeds were collected from 97 dairy farms in 56 towns of 13 wilayas (states/ provinces) of the central, eastern, western and southern regions. The samples were cultured on selective media and the obtained isolates were identified using bacteriological and molecular tests. Eighty-five Brucella isolates (72 B. abortus and 13 B. melitensis) were recovered from 63 animals in 37 dairy farms. In total, 71 (83.5 %) B. abortus bv 3, 11 (12.9 %) B. melitensis bv 2, 2 (2.4 %) B. melitensis bv 3 and 1 (1.2 %) unidentified B. abortus biovar were detected. The identification of B. abortus biovar 3 and B. melitensis biovar 2 is a new finding for Algeria and the Maghreb, respectively. B. abortus (84.7 %) was the main etiological agent of brucellosis. B. abortus showed a scattered distribution across Algeria. The fact that 60 % of the seropositive cattle showed no clinical signs, but 36 % were culture positive is an alarming observation. These data will rise awareness for the current epidemiological situation of bovine brucellosis in Algeria. To the best of our knowledge, this is the first representative countrywide bacteriological investigation of Brucella species and biovars in cattle across Algeria, which is a developing country where resources might be limited and the working conditions might not be very friendly.
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http://dx.doi.org/10.1016/j.vetmic.2021.109004DOI Listing
March 2021

Seroprevalence and Molecular Detection of Brucellosis in Hospitalized Patients in Lahore Hospitals, Pakistan.

Infect Dis Rep 2021 Feb 8;13(1):166-172. Epub 2021 Feb 8.

Friedrich-Loeffler-Institut, Institute of Bacterial Infections and Zoonoses, Naumburger Str. 96a, 07743 Jena, Germany.

Brucellosis is one of the most notorious zoonoses worldwide. The disease is common and endemic in humans and animals of Pakistan, but lack of awareness and lack of research have resulted in an increased incidence in the human population. The present study aimed to determine the seroprevalence and at molecular detection of brucellosis in patients with clinical symptoms in six different hospitals from Lahore, which is the capital city of Punjab province. A total of 218 blood samples were collected from hospitalized patients. The samples were initially screened by the Rose Bengal Plate Test (RBPT), and then quantitative real-time PCR (qRT-PCR) was applied. An overall seroprevalence of 17% (37/218) was found. The highest prevalence was found at the Lady Health center (36.53%), which was followed by the Lady Willingdon Hospital (28.6%). Female patients showed a higher seroprevalence than males and peaked at 34% (n = 32) for women who suffered from abortion. In total, 16.8% of patients younger than 30 years showed seropositive reactions, while the prevalence was 19% in patients between 31 and 50. Thirty-three DNA samples from 24 seropositive and nine seronegative patients tested positive, 32 samples were found positive for DNA, and one sample failed to be identified at the species level. Almost all positive cases had direct contact with animals and consumed unpasteurized dairy products. Research on human brucellosis is still scarce in Pakistan. For the diagnosis of brucellosis, serology and molecular tools should be combined if isolation by culture is not possible. Nationwide control activities and increasing awareness for zoonotic brucellosis are needed.
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http://dx.doi.org/10.3390/idr13010018DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7931085PMC
February 2021

The Animal-foods-environment interface of Klebsiella pneumoniae in Germany: an observational study on pathogenicity, resistance development and the current situation.

Vet Res 2021 Feb 8;52(1):16. Epub 2021 Feb 8.

Friedrich-Loeffler-Institut, Institute of Bacterial Infections and Zoonoses, Naumburger Street 96a, 07743, Jena, Germany.

Klebsiella (K.) pneumoniae as a multi-drug resistant (MDR) pathogen is an emerging challenge for clinicians worldwide. Virulence factors are capsular antigens, adherence factors, the O-lipopolysaccharide, and siderophores promoting infectivity. Mechanisms of antimicrobial resistance are inactivation of compounds via enzymes, change of membrane permeability, and alteration of the target site of the antimicrobial compound. In addition to environmental resistance, K. pneumoniae can survive increasing concentrations of disinfectants, if exposed. This review describes the temporal and spatial distribution of K. pneumoniae in the past decades in Germany, with emphases on the development of resistance in the non-human columns of the One-Health concept. In general, K. pneumoniae is a neglected pathogen in veterinary and environmental health, and the risk of human infection concerning animal contact and food consumption is barely investigated. Few reports exist (n = 26) on antibiotic resistance of isolates from non-human origin. Multi-drug resistance and extended-spectrum β-lactamase (MDR-ESBL) strains also resistant to carbapenems and antibiotics of the ß-lactam group harbor blaCTX-M, blaOXA, blaTEM, blaSHV, blaCMY, and PMQR have been found in animals, foods, and the environment. Colistin resistant strains carrying the mcr-1 gene were detected in wastewater. The blaCTX-M-15 and blaOXA-48 genes are the most frequently identified AMR genes in isolates of humans and were also the most predominant ESBL-genes in samples collected from animal hosts. Several aspects of the molecular epidemiology and resistance development of K. pneumoniae in farm animal populations, wildlife, and foods need intensive research. Environmental health has to be integrated into national research plans, as a lack of data is apparent. Increasing awareness of the fact that non-human sources can act as a reservoir for this pathogen has to be raised.
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http://dx.doi.org/10.1186/s13567-020-00875-wDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7871605PMC
February 2021

The Emergence of with Reduced Susceptibility Against Third Generation Cephalosporins and Carbapenems in Lagos Hospitals, Nigeria.

Antibiotics (Basel) 2021 Feb 1;10(2). Epub 2021 Feb 1.

Institute for Infectious Diseases and Infection Control, Jena University Hospital, Am Klinikum 1, 07747 Jena, Germany.

This study investigated the prevalence of (.) isolates among clinical samples of patients in four medical centers in Lagos, Nigeria and the burden of extended-spectrum beta-lactamases (ESBL) and carbapenem-resistant (CRKP) strains. Different samples (stool, blood, urine, wound swabs and nasal swabs) from 127 patients with suspected Gram-negative infections based on on-site performed Gram-stain from four public hospitals between March and September 2015 were analyzed. was identified in 43 (34%) patients. Resistance rates of these 43 strains according to the CLSI breakpoints were as followed: cotrimoxazole (90.7%), cefuroxime (74.4%), ofloxacin (55.8%), ceftazidime (46.5%), and cefixime (35%). Three isolates (7%) were resistant to imipenem. All isolates were susceptible to amoxicillin/clavulanic acid and nitrofurantoin. The prevalence of ESBL-producing, MDR and CRKP strains was 69.8%, 62.8%, and 7.0%, respectively. Of the ESBL-producing isolates, two isolates obtained from urine harbored both and , and a third isolate from urine harbored only the . This study revealed the emergence of CRKP isolates and and co-harboring strains in Lagos hospitals. The emergence of CRKP strains is an early warning signal for carbapenem antibiotics' prudent use with concern for their efficacies.
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http://dx.doi.org/10.3390/antibiotics10020142DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7912815PMC
February 2021

Characterization of Enterococci- and ESBL-Producing Isolated from Milk of Bovides with Mastitis in Egypt.

Pathogens 2021 Jan 21;10(2). Epub 2021 Jan 21.

Friedrich-Loeffler-Institut, Institute of Bacterial Infections and Zoonoses, Naumburger Str. 96a, 07743 Jena, Germany.

This study aimed to investigate the prevalence and antimicrobial resistance of enterococci- and ESBL-producing isolated from milk of bovine mastitis cases in Egypt. Fifty milk samples of dairy animals were collected from localities in the Nile Delta region of Egypt. Isolates were identified using MALDI-TOF MS, and antibiotic susceptibility testing was performed by the broth microdilution method. PCR amplifications were carried out, targeting resistance-associated genes. Seventeen isolates and eight coliform isolates could be cultivated. Vancomycin resistance rate was high in The VITEK 2 system confirmed all isolates as ESBL-producing. All . isolates harbored (B), L and D genes. The A gene was detected in isolate, B was found in other while one isolate of exhibited the A gene. isolates exhibited high prevalence of erm(B) and tetL. isolates were analyzed by DNA microarray analysis. Four isolates were determined by O-serotyping as O8 (n = 1), O86 (n = 2) and O157 (n = 1). H-serotyping resulted in H11, H12, H21 (two isolates each) and one was of H16 type. Different virulence-associated genes were detected in isolates including A, A, B, L, I1 and I2, and the N gene was identified by DNA microarray analysis.
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http://dx.doi.org/10.3390/pathogens10020097DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7909756PMC
January 2021

Seroprevalence and Molecular Identification of spp. in Bovines in Pakistan-Investigating Association With Risk Factors Using Machine Learning.

Front Vet Sci 2020 2;7:594498. Epub 2020 Dec 2.

Friedrich-Loeffler-Institut, Institute of Bacterial Infections and Zoonoses, Jena, Germany.

Bovine brucellosis is a global zoonosis of public health importance. It is an endemic disease in many developing countries including Pakistan. This study aimed to estimate the seroprevalence and molecular detection of bovine brucellosis and to assess the association of potential risk factors with test results. A total of 176 milk and 402 serum samples were collected from cattle and buffaloes in three districts of upper Punjab, Pakistan. Milk samples were investigated using milk ring test (MRT), while sera were tested by Rose-Bengal plate agglutination test (RBPT) and indirect enzyme-linked immunosorbent assay (i-ELISA). Real-time PCR was used for detection of DNA in investigated samples. Anti- antibodies were detected in 37 (21.02%) bovine milk samples using MRT and in 66 (16.4%) and 71 (17.7%) bovine sera using RBPT and i-ELISA, respectively. Real-time PCR detected DNA in 31 (7.71%) from a total of 402 bovine sera and identified as . Seroprevalence and molecular identification of bovine brucellosis varied in some regions in Pakistan. With the use of machine learning, the association of test results with risk factors including age, animal species/type, herd size, history of abortion, pregnancy status, lactation status, and geographical location was analyzed. Machine learning confirmed a real observation that lactation status was found to be the highest significant factor, while abortion, age, and pregnancy came second in terms of significance. To the authors' best knowledge, this is the first time to use machine learning to assess brucellosis in Pakistan; this is a model that can be applied for other developing countries in the future. The development of control strategies for bovine brucellosis through the implementation of uninterrupted surveillance and interactive extension programs in Pakistan is highly recommended.
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http://dx.doi.org/10.3389/fvets.2020.594498DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7738322PMC
December 2020

Revisiting Brucellosis in Small Ruminants of Western Border Areas in Pakistan.

Pathogens 2020 Nov 10;9(11). Epub 2020 Nov 10.

Institute of Bacterial Infections and Zoonoses, Friedrich-Loeffler-Institut, 07743 Jena, Germany.

Brucellosis, globally known bacterial zoonosis, is endemic to Pakistan. in bovines, in small ruminants and in dogs mainly cause this disease. A total of 1821 sera (1196 from sheep and 625 from goats) from animal herds near the Pakistan-Afghanistan border were collected. In parallel testing of sera for anti- antibodies ( and ) was carried out by RBPT and indirect ELISA. The presence of DNA in sera was tested by real-time PCR. The overall percentage of seropositive samples was 0.99 (18/1821) by both tests. All positive samples originated from Baluchistan territory which translated into 1.76% (18/1021). None of the positive sera had signals for DNA and none of sera from goats carried detectable antibodies. Both tests showed an almost perfect agreement with Kappa statistics. The flock size was found to be associated with the presence of anti- antibodies. The samples of Khyber Pakhtunkhwa (KPK) tested negative in both serological tests and hence were not processed for real-time PCR. The present study shows the presence of anti- antibodies in sheep in the Baluchistan region of Pakistan. Diagnostic services need to be improved and test and slaughter policies might be implemented for eradication of infection in these areas. Awareness about the infection is needed at the farmer's level. Isolation and molecular biology of the isolates could help with understanding the prevailing etiology in a better way.
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http://dx.doi.org/10.3390/pathogens9110929DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7697571PMC
November 2020

Epidemiology and Associated Risk Factors for Brucellosis in Small Ruminants Kept at Institutional Livestock Farms in Punjab, Pakistan.

Front Vet Sci 2020 2;7:526. Epub 2020 Sep 2.

Institute of Bacterial Infections and Zoonoses, Friedrich-Loeffler-Institut, Jena, Germany.

Brucellosis is reportedly endemic in ruminants in Pakistan. Both and infections have been decumented in domestic animals and humans in the country. This study aimed to identify the burden of anti- antibodies in small ruminants as well as associated potential risk factors with its occurrence at nine institutional livestock farms in Punjab, Pakistan. The sera collected from equal number of sheep and goats (500 from each species) were screened by indirect-ELISA for anti-smooth- antibodies followed by a serial detection by real-time PCR. Overall, 5.1% (51/1000) seropositivity was registered corresponding to 5% (25/500) prevalence in goats and 5.2% (26/500) in sheep. -DNA could not be detected in any of the tested sera by real-time PCR. Multiple logistic regression model indicated that farm location (OR 34.05), >4 years of age (OR 2.88), with history of reproductive disorders (OR 2.69), and with BCS of ≤ 3 (OR 12.37) were more likely to test positive for brucellosis at these farms. A routine screening, stringent biosecurity, and quarantine measures are warranted for monitoring and eradication of the infection. Similarly, isolation and molecular investigation of the etiologic agent(s) are needed to understand the relationship of epidemiology and out-breaks of brucellosis in the country.
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http://dx.doi.org/10.3389/fvets.2020.00526DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7493811PMC
September 2020

Prevalence and Spatial Distribution of Animal Brucellosis in Central Punjab, Pakistan.

Int J Environ Res Public Health 2020 09 21;17(18). Epub 2020 Sep 21.

Institute of Bacterial Infections and Zoonoses, Friedrich-Loeffler-Institut, Naumburger Str. 96a, 07743 Jena, Germany.

Brucellosis is an important zoonotic disease of animals and humans caused by bacteria of the genus . Brucellae are Gram-negative intracellular bacteria which infect a wide variety of animals including goats, sheep, buffaloes, cows, pigs, and wildlife. The objectives of this study were to determine the seroprevalence and spatial distribution of brucellosis in Central Punjab, Pakistan. A total of 1083 blood samples of goats, sheep, buffaloes, and cows were collected from 38 villages of four districts (Kasur, Faisalabad, Lahore, and Okara) of Punjab, Pakistan, and screened for brucellosis by Rose Bengal Plate test (RBPT) and PCR confirmed. Epidemiological, demographic data and GPS coordinates for every sample were collected. By using interpolation of the Aeronautical Reconnaissance Coverage Geographic Information System (Arc GIS), a surface plot was generated applying inverse distance weight (IDW). It was found that 35 (3.23%) serum samples were positive for brucellosis. In eight (61.5%), six (75%), seven (87.5%), and eight (89%) villages, positive goats, sheep, buffaloes, and cattle were detected, respectively. In general, older animals are more often positive for brucellosis. In goats bucks were more often RBPT positive than females while in sheep, buffaloes, and cattle more females were positive. The spatial distribution of brucellosis shows that it is widely distributed in the western region of the study area in goats and in the South-West region in sheep. Similarly, for buffaloes it is restricted to the south-east and north-west regions, and in cattle brucellosis is present in western region of study area only. Reflected by this study, brucellosis poses a risk for livestock in developing countries due to lack of awareness by officials, owners, and consumers, and control measures are missing. A risk map of brucellosis was generated to develop effective strategies for awareness rising and to improve the quality of control programs in Pakistan.
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http://dx.doi.org/10.3390/ijerph17186903DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7559556PMC
September 2020

Sheep Brucellosis in Kuwait: A Large-Scale Serosurvey, Identification of Species and Zoonotic Significance.

Vet Sci 2020 Sep 8;7(3). Epub 2020 Sep 8.

Department of Food Hygiene and Control, Faculty of Veterinary Medicine, Mansoura University, Mansoura 35516, Egypt.

Brucellosis is a common zoonotic disease of major concern in humans of Kuwait, and causes most human cases. The disease is endemic in small ruminants, cattle, and camels for decades, causing substantial economic losses in livestock production. However, a nationwide large-scale investigation of brucellosis in the small ruminant population has not been done in the past two decades. A serosurvey of sheep brucellosis in the five districts of Kuwait with most animal production farms was done between 2016 and 2019. In total, 67,054 serum samples from 233 sheep herds were collected and tested. Additionally, milk and tissue samples were collected from 46 seropositive cases for bacteriology. Thirty persons from seven seropositive farms were tested by serology. The incidence of seropositive cases was 7% in districts devoid of vaccination, while it was 4.7% in farms with history of vaccination. The serosurvey revealed that 89% of non-vaccinated herds ( = 181) were seropositive by Rose Bengal test (RBT), buffered acidified plate antigen test (BAPAT), and complement fixation test (CFT). Prevalence of 100% was reported for non-vaccinated sheep herds from Al-Wafrah and Al-Jahra districts, followed by those from Al-Salmi (88.24%), Al-Abdali (86.7%) and Kabd (75.6%). Implementation of vaccination with B. melitensis Rev.1 vaccine and test-and-slaughters in 20 herds reduced the seroprevalence to 33.3% and 25% in herds from Al-Jahra and AL-Wafrah, respectively. was isolated from 20 samples (43.5%). More than half of the examined animal owners (56.6%) tested positive for using RBT, BAPAT and CFT. The high numbers of infected herds and high prevalence in herdsmen are alarming. Thus, control measures have to be ensured immediately. The epidemiological situation in Kuwait is similar to those of the neighboring countries and the combined action of these states is needed. The understanding of the economic and public health impact of brucellosis in Kuwait needs to grow.
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http://dx.doi.org/10.3390/vetsci7030132DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7559804PMC
September 2020

Serological and Molecular Investigation of Brucellosis in Breeding Equids in Pakistani Punjab.

Pathogens 2020 Aug 19;9(9). Epub 2020 Aug 19.

Institute of Bacterial Infections and Zoonoses, Friedrich-Loeffler-Institut, 07743 Jena, Germany.

Brucellosis is an important zoonosis worldwide. Equines are susceptible to the infection when in close contact with infected animals. The objective of our study was to update the existing knowledge and detect and differentiate the causative agent of brucellosis in breeding equines in Punjab, Pakistan. A cross-sectional study was designed to evaluate the occurrence and etiology of the infection in the equine population in three districts. A total of 448 equine sera were collected from three prefectures viz. Sahiwal, Khanewal, and Okara of the Punjab Province of Pakistan. Ninety-six (21.4%) samples were found positive by RBPT, 3.56% (16/448) by iELISA, and 4.24% (19/448) by CFT. Real-time PCR demonstrated the presence of -DNA in sero-positive samples. Age and location were found as risk factors. The study concludes equine brucellosis seroprevalence in the country where as the main etiology. Fistulous withers and poll evil cases should be treated with care as they could be hazardous and a source of zoonotic transmission. Routine screening at an early age, vaccination in ruminants, and consumption of pasteurized dairy milk in humans is recommended for prevention of the infection. Specific tests need to be standardized and validated.
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http://dx.doi.org/10.3390/pathogens9090673DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7560188PMC
August 2020

Evaluation of the Diagnostic Potential of Recombinant Com1 in an ELISA for the Diagnosis of Q Fever in Sheep, Goats and Cattle.

Microorganisms 2020 Aug 13;8(8). Epub 2020 Aug 13.

Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Bacterial Infections and Zoonoses, 07743 Jena, Germany.

is the causative agent of Q fever, a zoonosis infecting domestic ruminants and humans. Currently used routine diagnostic tools offer limited sensitivity and specificity and symptomless infected animals may be missed. Therefore, diagnostic tools of higher sensitivity and specificity must be developed. For this purpose, the outer membrane protein Com1 was cloned and expressed in . The His-tagged recombinant protein was purified and used in an indirect enzyme-linked immunosorbent assay (ELISA). Assay performance was tested with more than 400 positive and negative sera from sheep, goats and cattle from 36 locations. Calculation of sensitivity and specificity was undertaken using receiver operating characteristic (ROC) curves. The sensitivities and specificities for sheep were 85% and 68% (optical density at 450nm, OD cut-off value 0.32), for goats 94% and 77% (OD cut-off value 0.23) and for cattle 71% and 70% (OD cut-off value 0.18), respectively. These results correspond to excellent, outstanding and acceptable discrimination of positive and negative sera. In summary, recombinant Com1 can provide a basis for more sensitive and specific diagnostic tools in veterinary medicine.
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http://dx.doi.org/10.3390/microorganisms8081235DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7465334PMC
August 2020

Phenotypic and WGS-derived antimicrobial resistance profiles of clinical and non-clinical Acinetobacter baumannii isolates from Germany and Vietnam.

Int J Antimicrob Agents 2020 Oct 1;56(4):106127. Epub 2020 Aug 1.

Institute for Infectious Diseases and Infection Control, Jena University Hospital, Jena, Germany; Research Campus Infectognostics, Jena, Germany.

Objectives: This study aimed to combine in vitro phenotyping analysis and whole-genome-sequencing (WGS) to characterise the phenotype and genetic determinants associated with intrinsic resistance in 100 clinical and non-clinical Acinetobacter baumannii strains originating from Germany and Vietnam. Moreover, it aimed to assess whether powdered milk as a food source functions as a potential reservoir of antibiotic resistance and possesses similar antimicrobial resistance (AMR) genes as in clinical strains isolated from Germany.

Methods: Antimicrobial susceptibility testing was performed using the broth microdilution method and the minimum inhibitory concentration (MIC) was determined for 18 antibiotics. The WGS data from all isolates were mapped to intrinsic genes known to be associated with phenotypic AMR.

Results: The highest resistance frequency was observed for chloramphenicol (100%), followed by fosfomycin (96%) and cefotaxime (95%). The lowest resistant rates were observed for colistin (3%), trimethoprim/sulfamethoxazole (17%), tigecycline (19%), and amikacin (19%). Thirty-five percent of tested strains displayed resistance to at least one of the carbapenems. Resistance to fluoroquinolones, aminoglycosides, tigecycline, penicillins, trimethoprim/sulfamethoxazole, and fourth-generation cephalosporins was determined only in human strains. About one-quarter of isolates (24%) was multidrug-resistant (MDR) and all were of human origin. Among them, 16 isolates were extensively drug resistant (XDR) and 10 from those 16 isolates showed resistance to all tested antibiotics except colistin. In silico detection of intrinsic AMR genes revealed the presence of 36 β-lactamases and 24 non-β-lactamase resistance genes. Two colistin-resistant and 10 ertapenem-resistant strains were isolated from powdered milk produced in Germany. Thirty-eight AMR genes associated with resistance to antibiotics were found in isolates recovered from milk powder. Several resistance mechanisms towards many classes of antibiotics existed in A. baumannii including β-lactamases, multidrug efflux pumps and aminoglycoside-modifying enzymes.

Conclusion: The use of WGS for routine public health surveillance is a reliable method for the rapid detection of emerging AMR in A. baumannii isolates. Milk powder poses a risk to contain MDR Acinetobacter strains or resistance genes in Germany.
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http://dx.doi.org/10.1016/j.ijantimicag.2020.106127DOI Listing
October 2020

Seroprevalence and Molecular Identification of spp. in Camels in Egypt.

Microorganisms 2020 Jul 13;8(7). Epub 2020 Jul 13.

Friedrich-Loeffler-Institut, Institute of Bacterial Infections and Zoonoses, 07743 Jena, Germany.

Brucellosis is one of the most important worldwide zoonoses of many countries including Egypt. Camel brucellosis has not gained much attention in Egypt yet. This study is focused on the three governorates with the highest camel populations and the largest camel markets in the country to determine the disease seroprevalence and identify the species in local camel holdings. In total, 381 serum samples were collected from male and female camels from Giza, Aswan, and Al-Bahr Al-Ahmar (the Red Sea) governorates. Samples were serologically examined using the Rose-Bengal plate test (RBPT), indirect ELISA (i-ELISA), competitive ELISA (c-ELISA) and complement fixation test (CFT). antibodies were detected in 59 (15.5%), 87 (22.8%), 77 (20.2%) and 118 (31.0%) of sera by RBPT, i-ELISA, c-ELISA and CFT, respectively. Using real-time PCR, DNA was amplified in 32 (8.4%) seropositive samples including (25/32), (5/32) and (2/32), defining a complex epidemiological status. To the best of our knowledge, this is the first study reporting DNA in camel serum. The risk-associated factors including age, sex, breed and geographical distribution were statistically analyzed, showing non-significant association with seroprevalence. The results of this study will raise awareness for camel brucellosis and help develop effective control strategies.
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http://dx.doi.org/10.3390/microorganisms8071035DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7409340PMC
July 2020

MLVA-16 Genotyping of and Isolates from Different Animal Species in Egypt: Geographical Relatedness and the Mediterranean Lineage.

Pathogens 2020 Jun 22;9(6). Epub 2020 Jun 22.

Friedrich-Loeffler-Institut, Institute of Bacterial Infections and Zoonoses, Naumburger Str. 96a, 07743 Jena, Germany.

Brucellosis is a common zoonotic disease in Egypt. However, there are limited data available on the genetic diversity of brucellae circulating in Egypt and other Mediterranean areas. One hundred and nine (.) strains were isolated from different animal species in thirteen Egyptian governorates. Multi-locus variable number tandem repeats (VNTRs) analysis (MLVA-16) was employed to determine the geographical relatedness and the genetic diversity of a panel of selected Egyptian strains ( = 69), with strains originating from Italy ( = 49), Portugal ( = 52), Greece ( = 63), and Tunisia ( = 4). Egyptian strains clustered into two main clusters containing 21 genotypes. Egyptian strains clustered into three main clusters containing nine genotypes. The genotypes were irregularly distributed over time and space in the study area. Egyptian strains of showed MLVA-16 patterns closer to that of Italian strains. Egyptian strains isolated from cattle share the same genotype with strains from Portugal and similar to strains from Italy with low genetic diversity. Strains with similar MLVA patterns isolated from different governorates highlight the movement of the pathogen among governorates. Hence, it may also reflect the long endemicity of brucellosis in Egypt with earlier dispersal of types and great local genetic diversity. Open markets may contribute to cross-species transmission and dissemination of the new types nationwide. The presence of West Mediterranean lineages of and relatedness of strains from the studied countries is a result of the socio-historical connections among the Mediterranean countries. Transnational eradication of brucellosis in the Mediterranean basin is highly demanded.
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http://dx.doi.org/10.3390/pathogens9060498DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7350383PMC
June 2020

Pan-Proteomic Analysis and Elucidation of Protein Abundance among the Closely Related Species, and .

Biomolecules 2020 05 30;10(6). Epub 2020 May 30.

Friedrich-Loeffler-Institute, Institute of Bacterial Infections and Zoonoses, Naumburger Str. 96a, 07743 Jena, Germany.

Brucellosis is a zoonotic infection caused by bacteria of the genus . The species, and , major causative agents of human brucellosis, share remarkably similar genomes, but they differ in their natural hosts, phenotype, antigenic, immunogenic, proteomic and metabolomic properties. In the present study, label-free quantitative proteomic analysis was applied to investigate protein expression level differences. Type strains and field strains were each cultured six times, cells were harvested at a midlogarithmic growth phase and proteins were extracted. Following trypsin digestion, the peptides were desalted, separated by reverse-phase nanoLC, ionized using electrospray ionization and transferred into an linear trap quadrapole (LTQ) Orbitrap Velos mass spectrometer to record full scan MS spectra ( 300-1700) and tandem mass spectrometry (MS/MS) spectra of the 20 most intense ions. Database matching with the reference proteomes resulted in the identification of 826 proteins. The Cluster of Gene Ontologies of the identified proteins revealed differences in bimolecular transport and protein synthesis mechanisms between these two strains. Among several other proteins, antifreeze proteins, Omp10, superoxide dismutase and 30S ribosomal protein S14 were predicted as potential virulence factors among the proteins differentially expressed. All mass spectrometry data are available via ProteomeXchange with identifier PXD006348.
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http://dx.doi.org/10.3390/biom10060836DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7355635PMC
May 2020

Proteomics of : Technologies and Their Applications for Basic Research and Medical Microbiology.

Microorganisms 2020 May 20;8(5). Epub 2020 May 20.

Institute of Animal Hygiene and Environmental Health, Centre for Infectious Medicine, Freie Universität Berlin, Robert-von-Ostertag-Str. 7-13, 14163 Berlin, Germany.

Brucellosis is a global zoonosis caused by Gram-negative, facultative intracellular bacteria of the genus Brucella (B.). Proteomics has been used to investigate a few B. melitensis and B. abortus strains, but data for other species and biovars are limited. Hence, a comprehensive analysis of proteomes will significantly contribute to understanding the enigmatic biology of brucellae. For direct identification and typing of Brucella, matrix-assisted laser desorption ionization - time of flight mass spectrometry (MALDI - TOF MS) has become a reliable tool for routine diagnosis due to its ease of handling, price and sensitivity highlighting the potential of proteome-based techniques. Proteome analysis will also help to overcome the historic but still notorious Brucella obstacles of infection medicine, the lack of safe and protective vaccines and sensitive serologic diagnostic tools by identifying the most efficient protein antigens. This perspective summarizes past and recent developments in Brucella proteomics with a focus on species identification and serodiagnosis. Future applications of proteomics in these fields are discussed.
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http://dx.doi.org/10.3390/microorganisms8050766DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7285364PMC
May 2020

Characterization of Staphylococci and Streptococci Isolated from Milk of Bovides with Mastitis in Egypt.

Pathogens 2020 May 15;9(5). Epub 2020 May 15.

Friedrich-Loeffler-Institut, Institute of Bacterial Infections and Zoonoses, Naumburger Str. 96a, 07743 Jena, Germany.

The aim of this study was to characterize staphylococci and streptococci in milk from Egyptian bovides. In total, 50 milk samples were collected from localities in the Nile Delta region of Egypt. Isolates were cultivated, identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), and antibiotic susceptibility testing was performed by the broth microdilution method. PCR amplifications were carried out, targeting resistance-associated genes. Thirty-eight isolates and six isolates could be cultivated. isolates revealed a high resistance rate to penicillin, ampicillin, clindamycin, and erythromycin. The A gene defining methicillin-resistant , (C) and D genes was found in 87.5% of each. Coagulase-negative staphylococci showed a high prevalence of A, Z and K genes. Other resistance-associated genes were found. All isolates carried Z, (A), (B), (C) and A genes, while harbored (C), A-3L and M genes, additionally. In , most of these genes were found. The isolate harbored Z, (B), (C), A, K, L and M genes. isolate was analyzed by DNA microarray analysis. It was determined as sequence type 14, belonging to clonal complex 19 and represented capsule type VI. Pilus and cell wall protein genes, A, D and B/A genes were identified by microarray analysis.
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http://dx.doi.org/10.3390/pathogens9050381DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7281669PMC
May 2020

Methicillin-Resistant (MRSA) in Poultry Species in Algeria: Long-Term Study on Prevalence and Antimicrobial Resistance.

Vet Sci 2020 Apr 27;7(2). Epub 2020 Apr 27.

Friedrich-Loeffler-Institut, Institute of Bacterial Infections and Zoonoses (IBIZ), Naumburger Str. 96a, 07743 Jena, Germany.

Methicillin-resistant (MRSA) is a well-known pathogen with a serious impact on human and veterinary public health. To determine antibiotic resistance of MRSA in poultry, 4248 nasal swabs were collected from 840 poultry farms in 18 different Wilayas (provinces) of Algeria. Swabs were collected between 2011 and 2018 from breeding hens, laying hens, broilers, and turkeys. Identification was carried out by the classical culture methods, and the disc diffusion test was used to determine the antibiotic resistance patterns. was isolated from 477 (56.8%) farms, and flock prevalence was 52.8%, 48.8%, 48.4%, and 75.6% in breeding hens, laying hens, broilers, and turkeys, respectively. MRSA was isolated from 252 (30%) farms and flock prevalence was 22%, 33.5%, 27.4%, and 36%, respectively. As expected, all MRSA isolates were resistant to cefoxitin, penicillin G, amoxicillin/clavulanic acid, and oxacillin. High levels of resistance were found for tetracycline (82.5%), erythromycin (70.6%), clindamycin (68.6%), and ciprofloxacin (50%). Almost all isolates were susceptible to vancomycin (100%) and mupirocin and rifampicin (99.2%), followed by chloramphenicol (82.3%), and gentamicin (76%). This moderate proportion of MRSA in poultry poses a considerable risk to public health. The results of this study highlight the need for control programs that encompass primary animal production and the food chain to mitigate contamination and spread of MRSA in the poultry industry of Algeria, and consequently to humans.
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http://dx.doi.org/10.3390/vetsci7020054DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7356745PMC
April 2020

Spatio-Temporal Distribution of in Germany-A Comprehensive Systematic Review of Studies on Resistance Development in Humans (2000-2018).

Microorganisms 2020 Mar 6;8(3). Epub 2020 Mar 6.

Institute for Infectious Diseases and Infection Control, Jena University Hospital, Am Klinikum 1, 07747 Jena, Germany.

() has gained global notoriety as a significant nosocomial pathogen because it is frequently associated with multi-drug resistance and hospital-based outbreaks. There is a substantial difference in the incidence of infections between different countries and within Germany. However, its continuous spread within Germany is a matter of concern. A systematic literature search and analysis of the literature published between 2000 and 2018 on in humans was performed. Forty-four studies out of 216 articles met the criteria for inclusion, and were selected and reviewed. The number of published articles is increasing over time gradually. Case reports and outbreak investigations are representing the main body of publications. North Rhine-Westphalia, Hesse and Baden-Wuerttemberg were states with frequent reports. Hospitals in Cologne and Frankfurt were often mentioned as specialized institutions. Multiresistant strains carrying diverse resistance genes were isolated in 13 of the 16 German states. The oxacillinase OXA-23-like, intrinsic OXA-51-like, OXA-58 variant, NDM-1, GES-11, CTX-M and TEM are the most predominant resistance traits found in German annii isolates. Five clonal lineages IC-2, IC-7, IC-1, IC-4 and IC-6 and six sequence types ST22, ST53, ST195, ST218, ST944/ST78 and ST348/ST2 have been reported. Due to multidrug resistance, colistin, tigecycline, aminoglycosides, fosfomycin, ceftazidime/avibactam and ceftolozan/tazobactam were often reported to be the only effective antibiotics left to treat quadruple multi-resistant Gram-negative (4MRGN) Dissemination and infection rates of are on the rise nationwide. Hence, several aspects of resistance development and pathogenesis are not fully understood yet. Increased awareness, extensive study of mechanisms of resistance and development of alternative strategies for treatment are required. One-Health genomic surveillance is needed to understand the dynamics of spread, to identify the main reservoirs and routes of transmission and to develop targeted intervention strategies.
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http://dx.doi.org/10.3390/microorganisms8030375DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7143851PMC
March 2020

Performance Analysis of Antibody Competitive ELISA Using the ROC Curve for Screening of Anaplasmosis in Camel Populations in Egypt.

Pathogens 2020 Feb 27;9(3). Epub 2020 Feb 27.

Institute of Bacterial Infections and Zoonoses, Friedrich-Loeffler-Institut (Federal research institute for Animal Health), Jena 07743, Germany.

Anaplasmosis is a tick-born and potential zoonotic disease caused by () , , and . affecting bovines and camels causing significant economic losses. Camels as an integral part of the socio-economic lifestyle of nomads in semi-arid to arid ecosystems are prone to suffer from subclinical infections. This study aimed to determine the performance and adaptation of commercial competitive ELISA (cELISA) as a tool for screening the seroprevalence of anaplasmosis whitin the camel populations in Egypt. This study was based on the serological investigation of 437 camel sera collected between 2015 and 2016 during a Q fever prevalence study in Egypt using commercially available cELISA for the detection of antibodies specific for in bovine serum. The receiver operating characteristic (ROC) curve, an analysis method for optimizing cutoff values in cELISAs, was used to estimate the sensitivity and specificity using 76 true as serological positive ( = 7) and negative ( = 60) for antibodies. ROC curve analysis was done for 7 true positive and 60 true negative bovine samples and 7 true positive and 29 true negative camel samples serum. Real time PCR and/or conventional PCR was applied to confirm spp. specific-DNA in camel serum as an indication of a true positive and true negative for ROC analysis. Chi square analysis was performed to estimate the association between risk factors and anaplasmosis in camels. The cutoff value was determined as 0.42 ( value ≤ 0.001). Data simulation with randomly generated values revealed a cutoff value of 0.417 ( ≤ 0.001) with resulting 58.1% and 97.8% . Seven true positive and 29 true negative camel serum samples was confirmed by PCR. Using the estimated cut off, the seroprevalence in the Nile Valley and Delta and the Eastern Desert domain was 47.4% and 46.4%, respectively. The potential risk factors as domains and origin of animals were less significantly associated with the prevalence of anaplasmosis (domains: χ(2) = 41.8, value ≤ 0.001 and origin: χ(2) = 42.56, value ≤ 0.001). Raising awareness especially for veterinarians and animal owners will significantly contribute to the best understanding of anaplasmosis in camels in Egypt. Alternative () validation techniques and preliminary prevalence studies are mandatory towards the control of neglected anaplasmosis in the camel population.
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http://dx.doi.org/10.3390/pathogens9030165DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7157543PMC
February 2020

Phenotypic and Molecular Detection of Biofilm Formation in Isolated from Different Sources in Algeria.

Pathogens 2020 Feb 24;9(2). Epub 2020 Feb 24.

Institute of Bacterial Infections and Zoonoses, Friedrich-Loeffler-Institut, 07743 Jena, Germany.

is an opportunistic bacterium causing a wide variety of diseases. Biofilm formation of is of primary public and animal health concern. The purposes of the present study were to investigate the ability of isolated from animals, humans, and food samples to form biofilms and to screen for the presence of biofilm-associated and regulatory genes. In total, 55 isolated from sheep mastitis cases (n = 28), humans (n = 19), and from food matrices (n = 8) were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The ability of aureus for slime production and biofilm formation was determined quantitatively. A DNA microarray examination was performed to detect adhesion genes (icaACD and biofilm-associated protein gene (bap)), genes encoding microbial surface components recognizing adhesive matrix molecules (MSCRAMMs), regulatory genes (accessory gene regulator (agr) and staphylococcal accessory regulator (sarA)), and the staphylococcal cassette chromosome mec elements (SCCmec). Out of 55 Staphylococcus aureus isolates, 39 (71.0%) and 23 (41.8%) were producing slime and biofilm, respectively. All strains isolated from food showed biofilm formation ability. 52.6% of the strains isolated from sheep with mastitis, and 17.9% of isolates from humans, were able to form a biofilm. Microarray analysis typed the Staphylococcus aureus into 15 clonal complexes. Among all isolates, four of the human isolates (21.1%) harbored the mecA gene (SCCmec type IV) typed into 2 clonal complexes (CC22-MRSA-IV and CC80-MRSA-IV) and were considered as methicillin-resistant, while two of them were slime-producing. None of the isolates from sheep with mastitis harbored the cna gene which is associated with biofilm production. The fnbB gene was found in 100%, 60% and 40% of biofilm-producing isolated from food, humans, and sheep with mastitis, respectively. Three agr groups were present and agr group III was predominant with 43.6%, followed by agr group I (38.2%), and agr group II (18.2%). This study revealed the capacity of Staphylococcus aureus isolates to form biofilms and highlighted the genetic background displayed by isolates from different sources in Algeria.
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http://dx.doi.org/10.3390/pathogens9020153DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7168657PMC
February 2020

Treatment of Yersinia similis with the cationic lipid DOTAP enhances adhesion to and invasion into intestinal epithelial cells - A proof-of-principle study.

Biochem Biophys Res Commun 2020 04 22;525(2):378-383. Epub 2020 Feb 22.

Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute for Bacterial Infections and Zoonoses, Naumburger Str. 96a, 07743 Jena, Germany. Electronic address:

The monocationic quaternary surfactant DOTAP has been used for the delivery of nucleic acids and peptides into mammalian cells. This study tested the applicability of DOTAP for the enhancement of adhesion and invasion frequencies of Yersinia (Y.) similis to enable the analysis of the effects of low-pathogenic bacteria on intestinal epithelial cells. Incubation of Y. similis with DOTAP ahead of infection of C2BBe1 intestinal epithelial cells increased invasion and adhesion frequency four- and five-fold, respectively, in plating assays. Proteomic approaches confirmed the increased bacterial load on infected cells: analysis of protein extracts by two-dimensional difference gel electrophoresis (2D-DIGE) revealed higher amounts of bacterial proteins present in the cells infected with DOTAP-treated bacteria. MALDI-TOF mass spectrometry of selected spots from gel-separated protein extracts confirmed the presence of both bacterial and human cell proteins in the samples. Label-free quantitative proteomics analysis identified 1170 human cell proteins and 699 bacterial proteins. Three times more bacterial proteins (279 vs. 93) were detected in C2BBe1 cells infected with DOTAP-treated bacteria compared to infections with untreated bacteria. Infections with DOTAP-treated Y. similis led to a significant upregulation of the stress-inducible ubiquitin-conjugating enzyme UBE2M in C2BBe1 cells. This points towards a stronger impact of the stress and infection responsive transcription factor AP-1 by enhanced bacterial load. DOTAP-treatment of uninfected C2BBe1 cells led to a significant downregulation of the transmembrane trafficking protein TMED10. The application of DOTAP could be helpful for investigating the impact of otherwise low adherent or invasive bacteria on cultivated mammalian cells without utilisation of genetic modifications.
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http://dx.doi.org/10.1016/j.bbrc.2020.02.081DOI Listing
April 2020