Clinical, pathologic, and molecular characterization of a non-metastatic multicentric cutaneous mast cell tumor in a cow.

Cutaneous mast cell tumors are rarely reported in cattle. Although mutations in the c-KIT gene have been shown to play a central role in the oncogenesis of canine mast cell tumors, few data are available in cattle. This report describes the clinical, histologic, immunohistochemical, and genetic features of a multicentric cutaneous mast cell tumor in an adult cow. An 11-year-old Prim'Holstein cow was presented for a 5-month history of multiple skin nodules. Cytologic and histologic analyses of the nodules led to a diagnosis of mast cell tumors. Immunohistochemical analysis for KIT expression showed a moderate to strong signal in neoplastic mast cells with a cytoplasmic and membranous pattern. Sequencing of the c-KIT gene coding sequence revealed no mutation. Despite partial response after corticosteroid treatment, euthanasia was elected. No metastases to the lymph nodes, spleen, and liver were identified at post-mortem and histologic examinations.

Surveillance of Extended-Spectrum ß-Lactamase-, Cephalosporinase- and Carbapenemase-Producing Gram-Negative Bacteria in Raw Milk Filters and Healthy Dairy Cattle in Three Farms in Île-de-France, France.

The aim of this work was to test a surveillance protocol able to detect extended-spectrum ß-lactamase (ESBL)-, cephalosporinase (AmpC)- and carbapenemase (CP)-producing gram-negative bacteria in three conveniently chosen dairy farms with known prior occurrences of ESBL- and CP-producing strains. The protocol was applied monthly for a year. At each visit, 10 healthy lactating dairy cows were rectally swabbed, and raw milk filters (RMFs) were sampled in two of the three farms. Bacterial isolation was based on a first screening step with MacConkey agar supplemented with 1 mg/L cefotaxime and commercial carbapenem-supplemented media. We failed to detect CP-producing strains but showed that ESBL-Escherichia strains, found in one farm only (13 strains), were closely associated with multi-drug resistance (12 out of 13). The limited number of conveniently selected farms and the fact that RMFs could not be retrieved from one of them limit the validity of our findings. Still, our results illustrate that ESBL-status changes monthly based on fecal swabs and negative herds should be qualified as "unsuspected" as proposed by previous authors. Although surveillance of farm statuses based on RMF analysis could theoretically allow for a better sensitivity than individual swabs, we failed to illustrate it as both farms where RMFs could be retrieved were constantly negative. Determination of CP herd-level status based on RMFs and our surveillance protocol was hindered by the presence of intrinsically resistant bacteria or strains cumulating multiple non-CP resistance mechanisms which means our protocol is not specific enough for routine monitoring of CP in dairy farms.

Cutaneous and systemic granulomatous disease associated with hairy vetch toxicosis in a French Holstein dairy herd.

This report describes an outbreak of hairy vetch toxicosis afflicting a herd of cattle with a fatal cutaneous and systemic granulomatous disease. It highlights how this condition remains poorly recognized by cattle production professionals in Europe and the need for communication about vetch-associated diseases.

Cet article décrit une épidémie de toxicose due à la vesce velue touchant un troupeau de bovins se manifestant par une maladie granulomateuse systémique et cutanée fatale. Ceci illustre comment cette atteinte reste peu connue par les professionnels de l'élevage en Europe et le besoin de communiquer sur les maladies associées à la vesce velue.

Este artículo describe un brote de toxicosis de veza vellosa (Vicia villosa) que afectó a un rebaño de ganado con una enfermedad granulomatosa cutánea y sistémica mortal. Destaca cómo esta condición sigue siendo poco reconocida por los profesionales de la producción ganadera en Europa y la necesidad de comunicación sobre las enfermedades asociadas al consumo de veza.

Este relato descreve um surto de intoxicação por ervilhaca peluda em um rebanho de gado com uma doença granulomatosa cutânea e sistêmica. Destaca-se como essa enfermidade continua pouco reconhecida pelos profissionais de bovinocultura na Europa, e a necessidade de comunicação sobre doenças associadas à ervilhaca.

Occurrence of Third-Generation Cephalosporins-Resistant Klebsiella pneumoniae in Fresh Fruits and Vegetables Purchased at Markets in Algeria.

This study aimed to characterize third-generation cephalosporin (3GC)-resistant Klebsiella pneumoniae isolated from fresh fruits and vegetables purchased at Bejaia city, Algeria. K. pneumoniae isolates were identified by MALDI-TOF. Susceptibility to antibiotics was tested by the disk diffusion method. Whole genome sequencing (WGS) was carried out to determine sequence type (ST), plasmid incompatibility group (Inc.), and acquired antimicrobial resistance gene presence. A total of 13 3GC-resistant K. pneumoniae strains were isolated. WGS identified blaCTX-M-15 in 11 extended-spectrum-beta-lactamases (ESBL)-K. pneumoniae and blaDHA-1 in 2 AmpC-K. pneumoniae. The aac(6')lb-cr gene was identified in 8 out of 13 isolates. Multilocus sequence typing (MLST) evidenced five different STs, namely ST14, ST45, ST219, ST236, and ST882. MDR K. pneumoniae contaminated fresh fruits and vegetables, often eaten raw and inappropriately washed, may represent an underestimated public health threat. This study highlights that hygiene measures during harvesting and retail process are of utmost importance to limit further ESBL/AmpC spread to the consumer households.

Functional Characterization of a Miniature Inverted Transposable Element at the Origin of mcr-5 Gene Acquisition in Escherichia coli.

Plasmid-mediated colistin resistance of the mobile colistin resistance (MCR) type is a growing concern in Enterobacteriaceae since it has been described worldwide in humans and animals. Here, we identified a series of MCR-producing Escherichia coli isolates corresponding to two different clones (represented by isolates PS1 and PS8b) producing MCR-1 and MCR-5, respectively, obtained from pig fecal samples in France. Plasmid analysis showed that the plasmid carrying the mcr-1 gene (pPS1) possesses an IncHI2 backbone, whereas the mcr-5 gene was carried onto a 6,268-bp nontypeable non-self-conjugative plasmid (pPS8b). Detailed analysis of plasmid pPS8b revealed a 3,803-bp-long cassette containing the mcr-5 gene that was bracketed by two inverted-repeat (IR) sequences with 5-bp-long direct repeats at each extremity, similarly to an insertion sequence, but with the exception that no transposase gene was identified within this cassette. By performing in vitro transposition experiments, we showed that the mcr-5 cassette could be mobilized by the TnAs1 transposase provided in trans, displaying a mobilization mechanism similar to that of miniature inverted-repeat transposable elements (MITEs).

From Farms to Markets: Gram-Negative Bacteria Resistant to Third-Generation Cephalosporins in Fruits and Vegetables in a Region of North Africa.

The role of food in human exposure to antimicrobial-resistant bacteria is a growing food safety issue. The contribution of fruits and vegetables eaten raw to this exposure is still unclear. The evaluation of contamination levels of fruits, vegetables and the agricultural environment by third-generation cephalosporin (3GC)-resistant Gram-negative bacteria was performed by analyzing 491 samples of fruits and vegetables collected from 5 markets and 7 farms in Bejaia area, north-eastern Mediterranean coast of Algeria. Ninety soil samples and 45 irrigation water samples were also sampled in farms in order to assess them as potential inoculum sources. All samples were investigated at the same time on ceftazidime-containing selective media for 3GC-resistant Gram-negative bacteria detection and on Hektoen media, for Salmonella spp. presence. The bacteria isolated (n = 30) from fruits and vegetables, soil and irrigation water collected in the farms were almost all non-fermenting bacterial species (Stenotrophomonas, Acinetobacter, Pseudomonas, Ochrobactrum) except one strain of Enterobacter cloacae and two strains of Citrobacter murliniae, isolated on one cucumber and two tomato samples in the same farm. Greater diversity in bacterial species and antimicrobial resistance profiles was observed at markets: Enterobacteriaceae (n = 41) were as strongly represented as non-fermenting bacteria (n = 37). Among Enterobacteriaceae, E. cloacae (n = 21), and Klebsiella pneumoniae (n = 13) were the most common isolates. Most of the K. pneumoniae isolates were extended-spectrum beta-lactamase (ESBL) producers (n = 11). No Salmonella spp. was recovered in any sample. This study showed that fruits and vegetables including those which may be eaten up raw constitute a reservoir of 3GC-resistant Gram-negative bacteria and multi-drug resistant-bacteria in general that can be transferred to humans through food. The general public should be informed of this hazard for health in order to encourage good domestic hygiene practices. In addition, further investigation is needed throughout the production chain to enrol professionals in actions to reduce this contamination.

Contribution of Avian Salmonella enterica Isolates to Human Salmonellosis Cases in Constantine (Algeria).

An epidemiological investigation was carried out on one hundred Salmonella isolates from broiler farms, slaughterhouses, and human patients in the Constantine region of Algeria, in order to explore the contribution of avian strains to human salmonellosis cases in this region over the same period of time. The isolates were characterized by phenotypic as well as genotypic methods. A large variety of antimicrobial resistance profiles was found among human isolates, while only seven profiles were found among avian isolates. Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR), Insertion Sequence 200-PCR (IS200-PCR), and Pulsed Field Gel Electrophoresis (PFGE) resulted in the allocation of the isolates to 16, 20, and 34 different profiles, respectively. The 3 genotyping methods led to complementary results by underlining the clonality of some serovars with the diffusion and persistence of a single clone in the Constantine area as well as stressing the polymorphism present in isolates belonging to other serovars, indicating the diversity of potential reservoirs of nontyphoidal Salmonella. Altogether, our results seem to indicate that nontyphoidal avian Salmonella may play an important role in human salmonellosis in the Constantine region.

Are Salmonella-Induced Gastroenteritis Neglected in Developing Countries? Feedback from Microbiological Investigations in N'Djamena Hospitals, Chad.

Salmonella is considered to be one of the main pathogens causing human gastroenteritis worldwide. Looking for Salmonella in Africa in patients suffering from gastroenteritis is rather unusual, and the use of antibiotics is not subject to any regulation. This study intends for stressing the possible prominent importance of Salmonella in digestive diseases in Africa as well as identifying antimicrobial resistance of Salmonella isolates from faeces samples of human origin. All samples were collected from five N'Djamena hospitals, from patients suffering from diarrhoea. The collecting was undertaken over two periods of six months each: from August 2010 to January 2011 and from September 2011 to February 2012. Salmonella isolates were obtained by standard cultivation and serotyping methods. A total of 43 Salmonella isolates were identified, belonging to 21 different serovars. The most prevalent serovar was Salmonella Stanleyville (n = 7), followed by S. Anatum (n = 4) and S. Kottbus (n = 3). The other serovars were under-represented. The majority of these isolates were susceptible to all antibiotics tested (CLSI Standards), except two S. Enteritidis isolates that exhibited resistance to fluoroquinolones. The different serovars and antibiotic resistance profiles that were observed highlight the substantial diversity of Salmonella in N'Djamena, Chad. Roughly, one out of ten patients who consulted for gastroenteritis was shedding Salmonella spp. and none of them would have been diagnosed outside the context of this research program. This study may encourage local clinicians to explore more often salmonellosis suspicion in their daily practice.

IncH-type plasmid harboring bla CTX-M-15, bla DHA-1, and qnrB4 genes recovered from animal isolates.

The whole sequence of plasmid pENVA carrying the extended-spectrum ß-lactamase gene blaCTX-M-15 was determined. It was identified from a series of clonally related Klebsiella pneumoniae sequence type 274 strains recovered from companion animals. This plasmid was 253,984 bp in size and harbored, in addition to blaCTX-M-15, a large array of genes encoding resistance to many antibiotic molecules, including ß-lactams (blaTEM-1, blaDHA-1), aminoglycosides (aacA2, aadA1), tetracycline (tetA), quinolones (qnrB4), trimethoprim (dfrA15), and sulfonamides (two copies of sul1). In addition, genes encoding resistance to mercury, tellurium, nickel, and quaternary compounds were identified. It also carried genes encoding DNA damage protection and mutagenesis repair and a locus for a CRISPR system, which corresponds to an immune system involved in protection against bacteriophages and plasmids. Comparative analysis of the plasmid scaffold showed that it possessed a structure similar to that of only a single plasmid, which was pNDM-MAR encoding the carbapenemase NDM-1 and identified from human K. pneumoniae isolates. Both plasmids possessed two replicons, namely, those of IncFIB-like and IncHIB-like plasmids, which were significantly different from those previously characterized. The blaCTX-M-15 gene, together with the other antibiotic resistance genes, was part of a large module likely acquired through a transposition process. We characterized here a new plasmid type carrying the blaCTX-M-15 gene identified in a K. pneumoniae isolate of animal origin. The extent to which this plasmid type may spread efficiently and possibly further enhance the dissemination of blaCTX-M-15 among animal and human isolates remains to be determined.

Prevalence and antimicrobial resistance of non-typhoidal Salmonella serotypes isolated from laying hens and broiler chicken farms in N'Djamena, Chad.

This study aimed at updating knowledge on the prevalence and antimicrobial resistance characteristics of Salmonella isolated from poultry in the province of N'Djamena, Chad. The results collected during this study provide the first baseline data on the prevalence of contamination by Salmonella in laying hens and broiler chicken farms in N'Djamena. All samples were collected from sixteen poultry farms over two periods of six months each: from August 2010 to January 2011 and from September 2011 to February 2012. Diagnostic methods used during this study allowed to isolate eighty four Salmonella strains, belonging to twenty seven different serotypes. The most frequent serotypes were Salmonella Colindale (19%) followed by S. Minnesota (18%) S. Havana and S. Riggil (each 6%), S. Kottbus and S. Amager (4.7%), S. Idikan, Mississipi, and Muenchen (3.6%). Other serotypes were poorly represented. The majority of these serotypes were susceptible to all antibiotics tested (CLSI Standards), except some S. Colindale isolates that exhibited a decreased susceptibility to fluoroquinolones, S. Limete resistant to three antibiotics and S. Minnesota isolates resistant to five different antimicrobial classes. The different serotypes and antibiotic resistance profiles that were observed highlight the substantial diversity of Salmonella in Chad, the contribution of avian isolates to human salmonellosis and Salmonella's capacity to colonize all types of environment worldwide.

Extended-spectrum ß-lactamase CTX-M-15-producing Klebsiella pneumoniae of sequence type ST274 in companion animals.

Screening of extended-spectrum ß-lactamase (ESBL)-producing Gram-negative bacteria in companion animals living in the Paris area in France identified a high rate of CTX-M-15-producing Klebsiella pneumoniae. Those isolates were recovered during the 2010-2011 period from both infections and asymptomatic colonizations. Sequence typing revealed that most of these isolates belonged to sequence type ST274. Interestingly, the bla(CTX-M-15) gene was located on a specific and novel plasmid scaffold. These findings highlight that companion animals may be reservoirs for CTX-M-15-producing K. pneumoniae evolving separately from the human reservoir of CTX-M-15 producers.

Ceftriaxone-resistant salmonella enterica serotype Newport, France.

The multidrug-resistant (MDR) Salmonella enterica serotype Newport strain that produces CMY-2 beta-lactamase (Newport MDR-AmpC) was the source of sporadic cases and outbreaks in humans in France during 2000-2005. Because this strain was not detected in food animals, it was most likely introduced into France through imported food products.

A comparison of antimicrobial usage in human and veterinary medicine in France from 1999 to 2005.

OBJECTIVES: The antimicrobials allowed and amounts sold in veterinary and human medicine in France were compared to see if the same antimicrobial drugs are used in veterinary and human medicine, and to the same extent. METHODS: Registers of all approved antimicrobial commercial products, kept by the French Agency for Veterinary Medicinal Products (AFSSA ANMV) for animals and the French Health Products Safety Agency (AFSSAPS) for humans, were compared to determine whether the same antimicrobials were approved in 2007 for use in both human and animal populations. Sales data were collected from pharmaceutical companies between 1999 and 2005 by the AFSSA ANMV and AFSSAPS. Usage of the different antimicrobial anatomical therapeutic chemical (ATC) classes in human and veterinary medicines was recorded. Data were expressed in tonnes of active ingredients and were then related to the animal and human biomasses to compare usages expressed in mg/kg. RESULTS: All antimicrobial ATC classes were used in both human and veterinary medicine. Tetracyclines accounted for the most sales in veterinary medicine. beta-Lactams predominated in human medicine. A decrease in the amounts consumed by both human and animal populations was observed during the study. In 2005, 760 tonnes were used in human medicine and 1320 tonnes in veterinary medicine, corresponding to 199 and 84 mg/kg of live weight in human and animal populations, respectively. CONCLUSIONS: The same antimicrobial drugs were used in human and veterinary medicines but the quantitative patterns of use were different. Expression of antimicrobial usage is a key point to address when comparing usage trends.

Epidemiological analysis of Salmonella enterica from beef sampled in the slaughterhouse and retailers in Dakar (Senegal) using pulsed-field gel electrophoresis and antibiotic susceptibility testing.

Seventy-eight isolates of Salmonella spp. isolated from beef sampled from the official city slaughterhouse and from retailers in Dakar, Senegal were analyzed using serotyping, antimicrobial testing and macrorestriction profiling by Pulsed-Field Gel Electrophoresis (PFGE). These analyses were done to identify clonal relationships and potential transmission routes in beef channel. XbaI macrorestriction allowed defining 17 genotypes among the six main analyzed serotypes: Salmonella bredeney (3 genotypes), S. muenster (6), S. waycross (1), S. corvallis (3), S. kentucky (1) and S. brandenburg (3). The cross analysis of PFGE profiles and origin of the beef samples reveals a wide range of contamination sources in the beef channel in Dakar. Comparison of PFGE and antimicrobial resistance types shows that the Salmonella contamination sources are equally shared by the slaughterhouse (56% of the isolates) and by the distribution channel (44% of the isolates) by handlings and houseflies.

Prevalence and antibiotic-resistance of Salmonella isolated from beef sampled from the slaughterhouse and from retailers in Dakar (Senegal).

A study was made of Salmonella contamination in beef sampled from a slaughterhouse and from retailers in Dakar, Senegal. The serotypes as well as antibiotic-resistance patterns of the Salmonella isolates were determined. A total of 435 meat samples (236 from the slaughterhouse, 199 from retailers) were tested. Among them, 275 (63%) were positive for Salmonella, 43% (101/236) from the slaughterhouse and 87% (174/199) from the retailers. Furthermore, 97% of the investigated retailers had at least one beef sample contaminated by Salmonella. The 286 Salmonella isolates were divided into 51 serotypes. The most prevalent serotypes were Salmonella bredeney (25%), S. muenster (8%), S. waycross (7%), S. corvallis (4%) and S. kentucky (4%). About 62% of the isolates were resistant to nitrofurans. Resistance rates were lower to streptomycin (22%), sulfamethoxazole (15%), spectinomycin (1%), chloramphenicol (1%), and tetracycline (0,4%) while low-level resistance to quinolones was detected. About 16% of the Salmonella strains were multiresistant to two or more antibiotic families. Finally, ten resistance profiles have been identified. This study shows the huge spread of Salmonella in the beef production chain in Dakar, Senegal. Finally, this study provides the very first data about Salmonella prevalence in sub-saharian Africa.

Prevalence of multidrug resistant (MDR) Salmonella in bovine dairy herds in western France.

As a part of our effort in quantitative risk analysis of food-borne diseases, we carried out an epidemiologic study to estimate the prevalence of multidrug resistant (MDR) Salmonella in dairy herds situated in western France. The study population consisted of 489 farms in the region and manure or slurry was sampled from these operations and tested for the Salmonella spp. All strains isolated during the study were serotyped and tested for their antimicrobial susceptibility. Salmonella spp. was isolated from 8.1% (95% confidence interval (CI 95%): 4.5-13.3%) of the sampled herds. The herd prevalence of MDR Salmonella among the sampled herds was 1.9% (CI 95%: 0.5-5.4%). Spatial statistics were used to check for sampling representativeness and to determine if infected herds were clustered spatially.