This synopsis summarizes the key chemical and bacteriological characteristics of β-lactams, penicillins, cephalosporins, carbanpenems, monobactams and others. Particular notice is given to first-generation to fifth-generation cephalosporins. This review also summarizes the main resistance mechanism to antibiotics, focusing particular attention to those conferring resistance to broad-spectrum cephalosporins by means of production of emerging cephalosporinases (extended-spectrum β-lactamases and AmpC β-lactamases), target alteration (penicillin-binding proteins from methicillin-resistant Staphylococcus aureus) and membrane transporters that pump β-lactams out of the bacterial cell.

Antibiotics have been intensively used over the last decades in human and animal therapy and livestock, resulting in serious environmental and public health problems, namely due to the antibiotic residues concentration in wastewaters and to the development of antibiotic-resistant bacteria. This study aimed to access the contribution of some anthropological activities, namely urban household, hospital and a wastewater treatment plant, to the spread of antibiotic resistances in the treated wastewater released into the Mondego River, Coimbra, Portugal. Six sampling sites were selected in the wastewater network and in the river. The ampicillin-resistant Enterobacteriaceae of the water samples were enumerated, isolated and phenotypically characterized in relation to their resistance profile to 13 antibiotics. Some isolates were identified into species level and investigated for the presence of class A and class C -lactamases. Results revealed high frequency of resistance to the -lactam group, cefoxitin (53.5%), amoxicillin/clavulanic acid combination (43.5%), cefotaxime (22.7%), aztreonam (21.3) cefpirome (19.2%), ceftazidime (16.2%) and to the non--lactam group, trimethoprim/sulfamethoxazol (21.1%), tetracycline (18.2%), followed by ciprofloxacin (14.1%). The hospital effluent showed the higher rates of resistance to all antibiotic, except two (chloramphenicol and gentamicin). Similarly, higher resistance rates were detected in the wastewater treatment plant (WWTP) effluent compared with the untreated affluent. Regarding the multidrug resistance, the highest incidence was recorded in the hospital sewage and the lowest in the urban waste. The majority of the isolates altogether are potentially extended-spectrum -lactamases positive (ESBL⁺) (51.9%), followed by AmpC⁺ (44.4%) and ESBL⁺/AmpC⁺ (35.2%). The most prevalent genes among the potential ESBL producers were bla_OXA (33.3%), bla_TEM (24.1%) and bla_CTX-M (5.6%) and among the AmpC producers were bla_EBC (38.9%), bla_FOX (1.9%) and bla_CIT (1.9%). In conclusion, the hospital and the WWTP activities revealed to have the highest contribution to the spread of multidrug resistant bacteria in the study area. Such data is important for future management of the environmental and public health risk of these contaminants. This is the first embracing study in the water network of Coimbra region on the dissemination of antibiotic resistance determinants. Moreover, it is also the first report with the simultaneous detection of multiresistant bacteria producers of AmpC and ESBLs -lactamases in aquatic systems in Portugal.

The exposure of both crop fields and humans to antibiotic-resistant bacteria in animal excreta is an emergent concern of the One Health initiative. This study assessed the contamination of livestock manure from poultry, pig, dairy farms and slaughterhouses in Portugal with resistance determinants. The resistance profiles of 331 Enterobacteriaceae isolates to eight β-lactam (amoxicillin, cefoxitin, cefotaxime, cefpirome, aztreonam, ceftazidime, imipenem and meropenem) and to five non-β-lactam antibiotics (tetracycline (TET), trimethoprim/sulfamethoxazole (SXT), ciprofloxacin (CIP), chloramphenicol (CHL) and gentamicin) was investigated. Forty-nine integron and non-β-lactam resistance genes were also screened for. Rates of resistance to the 13 antibiotics ranged from 80.8% to 0.6%. Multidrug resistance (MDR) rates were highest in pig farm samples (79%). Thirty different integron and resistance genes were identified. These were mainly associated with resistance to CHL (catI and catII), CIP (mainly, qnrS, qnrB and oqx), TET (mainly tet(A) and tet(M)) and SXT (mostly dfrIa group and sul3). In MDR isolates, integron presence and non-β-lactam resistance to TET, SXT and CHL were positively correlated. Overall, a high prevalence of MDR Enterobacteriaceae was found in livestock manure. The high gene diversity for antibiotic resistance identified in this study highlights the risk of MDR spread within the environment through manure use.

Extended-spectrumβ-lactamases (ESBLs) prevalence was studied in the north of Portugal, among 193 clinical isolates belonging to citizens in a district in the boundaries between this country and Spain from a total of 7529 clinical strains. In the present study we recovered some members of Enterobacteriaceae family, producing ESBL enzymes, includingEscherichia coli(67.9%),Klebsiella pneumoniae(30.6%),Klebsiella oxytoca(0.5%),Enterobacter aerogenes(0.5%), andCitrobacter freundii(0.5%).β-lactamases genes blaTEM, blaSHV, and blaCTX-M were screened by polymerase chain reaction (PCR) and sequencing approaches. TEM enzymes were among the most prevalent types (40.9%) followed by CTX-M (37.3%) and SHV (23.3%). Among our sample of 193 ESBL-producing strains 99.0% were resistant to the fourth-generation cephalosporin cefepime. Of the 193 isolates 81.3% presented transferable plasmids harboringblaESBLgenes. Clonal studies were performed by PCR for the enterobacterial repetitive intragenic consensus (ERIC) sequences. This study reports a high diversity of genetic patterns. Ten clusters were found forE. coliisolates and five clusters forK. pneumoniaestrains by means of ERIC analysis. In conclusion, in this country, the most prevalent type is still the TEM-type, but CTX-M is growing rapidly.

The purpose of this study was to investigate the presence of β-lactam-resistant bacteria in six different types of Portuguese cheese. The numbers of ampicillin resistant (AMP^r) bacteria varied from 4.7 x 10² to 1.5 x 10⁷ CFU/g. Within 172 randomly selected β-lactam-resistant bacteria, 44 resistant phenotypes were found and 31.4% were multidrug resistant. The majority (85%) of the isolates identified belonged to the Enterobacteriaceae family. The presence of the bla_TEM gene was detected in 80.9% of the tested isolates. The results suggest that without thermal processing of the milk and good hygienic practices, cheese may act as a vehicle of transfer of β-lactam-resistant bacteria to the gastrointestinaltract of consumers.

Irrigation districts play a decisive role in Portuguese agriculture and require the adaptation to the new water management paradigm through a change in technology and practices compatible with farmers’ technical know-how and economic sustainability. Therefore, improvement of water management, focusing on water savings and increasing farmers’ income, is a priority. In this perspective, an applied research study is being carried out on the gravity-fed Lis Valley Irrigation District to assess the performance of collective water supply, effectiveness of water pumping, and safety of crop production due to the practice of reuse of drainage water. The water balance method was applied at irrigation supply sectors, including gravity and Pumping Irrigation Allocation. The average 2018 irrigation water allocated was 7400 m³/ha, being 9.3% by pumping recharge, with a global efficiency of about 67%. The water quality analysis allowed identifying some risk situations regarding salinization and microbiological issues, justifying action to solve or mitigate the problems, especially at the level of the farmers’ fields, according to the crops and the irrigation systems. Results point to priority actions to consolidate improved water management: better maintenance and conservation of infrastructure of hydraulic infrastructures to reduce water losses and better flow control; implementation of optimal operational plans, to adjust the water demand with distribution; improvement of the on-farm systems with better water application control and maintenance procedures; and improvement of the control of water quality on the water reuse from drainage ditches. The technological innovation is an element of the modernization of irrigation districts that justifies the development of multiple efforts and synergies among stakeholders, namely farmers, water users association, and researchers.