Aims Molecular characterization of extended-spectrum β-lactamases (ESBLs) among Salmonella Kentucky and Typhimurium isolates: partial sequence analysis of the types of β-lactamases found in these isolates, clonality, resistance and supposed emergence of ESBL-producing strains. Methods and Results A retrospective study surveyed the ESBLs occurring in a total of 1404 Salmonella Kentucky and Typhimurium isolates collected over a 5-year period in Tunisia. Antimicrobial susceptibility tests, ESBL phenotype determination (double-disc synergy) were performed. Polymerase chain reaction assays were used for the detection of β-lactamase genes (bla_TEM, bla_SHV, bla_OXA-1 and bla_CTX-M), class 1 and class 2 integrases (intI1 and intI2) and the 3′ conserved segment (3′-CS) of class 1 integron (qacEΔ1+sul1). Sequencing of amplicons of β-lactamase genes was performed. Percentage of 9.8 of the isolates (S. Kentucky = 117, S. Typhimurium = 20) were either resistant to penicillin and had decreased susceptibility to cefotaxime or had a positive double-disc synergy test result. Polymerase chain reaction detected that these isolates harboured one or more β-lactamase genes (bla_TEM, bla_SHV, bla_OXA-1 or bla_CTX-M). TEM-1, TEM-34, CTX-M15, CTX-M9 and CTX-M61 type ESBLs were identified through sequencing. The novel Salmonella cefotaxime-hydrolysing β-lactamase, CTX-M61/TEM-34, detected in this study showed the emergence of new CTX-M-type ESBLs in Tunisia. There were found 33 different multidrug resistance (MDR) patterns. Conclusion These findings highlighted the proliferation of ESBLs and MDR in Salmonella Kentucky and Typhimurium isolates from numerous regions and sources in Tunisia, indicating an emerging public health concern. Significance and Impact of the Study For the first time CTX-M-61/TEM-34, a novel cefotaxime-hydrolysing β-lactamase of Salmonella had been detected.

Aims Characterization of quinolone‐resistant Salmonella Kentucky and Typhimurium isolates in Tunisia from various sources, detection of some PMQR genes, and the genetic relatedness. Methods A total of 1,404 isolates of S . Kentucky (n = 1,059) / S . Typhimurium (n = 345) from various sources from all over Tunisia were tested for quinolone resistance by disk diffusion method. Minimum inhibitory concentrations of nalidixic acid, ciprofloxacin, and ofloxacin were determined. Quinolone‐resistant isolates were screened for plasmid‐mediated quinolone‐resistance genes [qnrA, qnrB, qnrS , aac(6′)‐Ib‐cr , and qepA ] by PCR. Mutations in the quinolone resistance‐determining regions of the gyrA and parC genes were detected by PCR and DNA sequencing. Pulsed‐field gel electrophoresis and multilocus sequence typing were accomplished for isolates harboring plasmid‐mediated quinolone‐resistance genes. Results According to our selection criteria (NAL = resistance phenotype; CIP = resistant with diameter 0, or intermediate), only 63 S. Kentucky/41 S . Typhimurium isolates were investigated: 49% (5/104) were multidrug‐resistant. Two S . Typhimurium isolates harbored qnrB19 with different PFGE profiles. A mutation was detected in the gyrA gene for each of these two isolates. MLST revealed the presence of ST313 and ST34, an endemic sequence type. Conclusion Our study highlights the presence of quinolone multidrug‐resistant Salmonella in humans and animals in Tunisia. This is the first report of S. Typhimurium ST34 in Africa and qnrB19 in Tunisia.

The poultry industry is a very important agricultural and industrial sector in Tunisia and Nigeria, with little information about occurrence of diarrheagenic Escherichia coli in the farmers and chickens. This study aimed to detect the prevalence of diarrheal E. coli in humans and poultry and to investigate plasmid-mediated quinolone resistance (PMQR) genes in both countries. Seventy-four isolates of E. coli were studied; nine different virulence genes were screened by PCR. Serotyping was performed only for pathotypes as well as the determining of antibiotic resistance profiles against 21 antibiotics. PMQR genes were investigated by PCR. EAEC was the most abundant pathotype (37/74; 50%) in human and chicken isolates, whereas single EHEC and EPEC (1/74, 1.35%) pathotypes were detected in Tunisia and Nigeria, respectively. About 17 (45.95%) quinolones/fluoroquinolones-resistant isolates were detected, from which the following PMQR genes were detected: aac(6′)-Ib-cr (8/17, 47.05%), qepA (6/17, 35.29%), qnrA + qnrB (2/17, 11.76%), and qnrS gene (1/17, 5.88%). Our findings highlight high occurrence of EAEC pathotype in Tunisia and Nigeria, more frequent than EPEC and EHEC. Additionally, all E. coli pathotypes isolated from different sources (humans, poultry) showed resistance to several antibiotics, which are in use as therapeutic choices in Tunisia and Nigeria.

Background and Aim: Brucellosis contributes to significant economic losses due to abortion, weak newborns, infertility, and up to 20% reductions in milk yield in carrier animals. This study aimed to estimate the prevalence of ruminant brucellosis in six Egyptian governorates. This study aimed to estimate the prevalence of ruminant brucellosis and evaluate the risk factors regarding the epidemiological status, highlighting the importance of early carrier detection for the success of control programs. Materials and Methods: A total of 3000 ruminants were investigated. Blood and serum samples were collected for routine hemato-biochemical analysis (complete blood picture and metabolic panel). In addition, genotoxicity analysis was performed, whereas tissue samples were collected for histopathological analysis. The buffered acidified plate antigen test (BAPAT), Rose Bengal plate test (RBPT), and complement fixation test (CFT) were used for serological diagnosis of brucellosis. The obtained bacterial colonies were typed using Brucella abortus-, melitensis-, ovis-, and suis-polymerase chain reaction (AMOS-PCR), depending on the variability of the IS711 fragment among Brucella spp. Serum trace elements, oxidative stress, and acute phase proteins were compared according to body condition score (BCS) and clinical condition images within the study population. Results: Mastitis and abortion were the key recorded symptoms (9.966%, 299/3000 and 6%, 180/3000, respectively); however, symptomless individuals were predominant (82.9%, 2487/3000). Blood lymphocytosis was prominent even in asymptomatic animals. Nutritional and food conversion conditions were defined as low, moderate, or high BCS. Brucella overall seropositivity by BAPAT, RBPT, and CFT was 6.1% (182/3000), 5.6% (168/3000), and 5.1% (154/3000) in ruminant species within the included governorates, respectively. Upon diagnosis, 154 seropositive cases developed 93 bacterial isolates and a 731-bp PCR fragment whose sequences confirmed Brucella melitensis biovar 3. Serum metabolic and biochemical profiles, acute phase proteins, trace elements, and oxidative stress concentrations were indicative of loss of functionality in the liver and kidneys, malnutrition and malabsorption syndrome, and DNA damage, particularly in the low-BCS groups (p < 0.0001). Granulomatous lesions were most prominent in the lymph nodes, spleen, uterus, and udder of the dams, while placental multifocal necrosis with thrombosis was recorded in aborted fetuses. There were 8 types of chromosomal aberrations detected in peripheral white blood cells. The highest frequency was for dicentric aberrations 0.025% (25/1000), whereas the lowest 0.009% (9/1000) was for acentric, ring, fusion, and polyploidy. The difference between species was significant for BCS; 14.2% in low-BCS cattle and camels and 8.4% in high-BCS buffaloes. Conclusion: B. melitensis biovar 3 is prevalent in Egypt. Mixed-rearing systems are the main risk factors for interspecies transmission among ruminants. The difficulty in accurately diagnosing all infected animals, particularly carriers, is a major limitation of eradication and control programs. Different biomarkers could be indicators and/or sensors for performance and/or infectivity conditions in animal herds; however, they require further optimization. Early detection using molecular technologies, highly descriptive, quantitative, sensitive, and specific methods, as alternatives to serological diagnosis (CFT, BAPAT, and RBT), is urgently needed to enhance the efficiency of brucellosis-specific prophylaxis. Such a comprehensive procedure is the World Organization for Animal Health dependent decision. Keywords: abortion, acute phase proteins, AMOS-PCR, body condition score, Brucella melitensis, Egypt, genotoxicity, mastitis, oxidative stress, prevalence, trace elements.

Aquatic ecosystems are recognized as major reservoirs of antimicrobial resistance (AMR), where chemical contaminants interact with microbial populations to promote the persistence and spread of res...