ABSTRACT
According to the 2030 Agenda of the United Nations, one of the sustainable development goals is to ensure sustainable consumption and production patterns. The need to ensure food safety includes, other than microbiological hazards, concerns with antimicrobial-resistant (AMR) bacteria. The emergence of resistant bacteria in the food industry is essentially due to the abusive, and sometimes incorrect, administration of antimicrobials. Although not allowed in Europe, antimicrobials are often administered to promote animal growth. Each time antimicrobials are used, a selective pressure is applied to AMR bacteria. Moreover, AMR genes can be transmitted to humans through the consumption of meat-harbouring-resistant bacteria, which highlights the One Health dimension of antimicrobial resistance. Furthermore, the appropriate use of antimicrobials to ensure efficacy and the best possible outcome for the treatment of infections is regulated through the recommendations of antimicrobial stewardship. The present manuscript aims to give the current state of the art about the transmission of AMR bacteria, particularly methicillin-resistant S. aureus, ESBL-producing Enterobacteriaceae, and vancomycin-resistant Enterococcus spp., along with other ESKAPE bacteria, from animals to humans through the consumption of meat and meat products, with emphasis on pork meat and pork meat products, which are considered the most consumed worldwide.
ABSTRACT
Antimicrobial resistance is a serious problem for the control of infections and infectious diseases. Propolis is a substance produced by honeybees with antimicrobial and antibiofilm properties. To consider propolis as an alternative to the use of antimicrobials for infection control, we assessed its antimicrobial and antibiofilm activities. To assess propolis for topical medical use, toxicological studies were also performed. A Portuguese 70% propolis ethanolic extract was chemically evaluated and studied for antimicrobial activity on staphylococcal field isolates (n = 137) and antibiofilm action (n = 45). Cell toxicological assessment was performed using keratinocytes and fibroblasts. Pinobanksin, chrysin, acacetin, apigenin, pinocembrin, and kaempferol-dimethyl-ether were detected. All 137 isolates were susceptible to 6.68 mg/mL or lower propolis concentration (80% isolates were susceptible to <1 mg/mL). The mean percentage of biofilm inhibition was 71%, and biofilm disruption was 88.5%. Propolis (<1 mg/mL) was well-tolerated by fibroblasts and moderately tolerated by keratinocytes. The combined antimicrobial and antibiofilm effect of propolis, together with its low toxicity to connective tissue and epithelial cells, suggests a good applicability for topical antibacterial treatment. Therefore, propolis seems to be a good alternative to antimicrobials for the treatment of infections with Staphylococcus spp. that deserves to be evaluated in vivo.
ABSTRACT
Small ruminant mastitis is a serious problem, mainly caused by Staphylococcus spp. Different virulence factors affect mastitis pathogenesis. The aim of this study was to investigate virulence factors genes for biofilm production and antimicrobial resistance to ß-lactams and tetracyclines in 137 staphylococcal isolates from goats (86) and sheep (51). The presence of coa, nuc, bap, icaA, icaD, blaZ, mecA, mecC, tetK, and tetM genes was investigated. The nuc gene was detected in all S. aureus isolates and in some coagulase-negative staphylococci (CNS). None of the S. aureus isolates carried the bap gene, while 8 out of 18 CNS harbored this gene. The icaA gene was detected in S. aureus and S. warneri, while icaD only in S. aureus. None of the isolates carrying the bap gene harbored the ica genes. None of the biofilm-associated genes were detected in 14 isolates (six S. aureus and eight CNS). An association was found between Staphylococcus species and resistance to some antibiotics and between antimicrobial resistance and animal species. Nine penicillin-susceptible isolates exhibited the blaZ gene, questioning the reliability of susceptibility testing. Most S. aureus isolates were susceptible to tetracycline, and no cefazolin or gentamycin resistance was detected. These should replace other currently used antimicrobials.
ABSTRACT
OBJECTIVES: Mastitis is responsible for a decrease in milk yield and quality. Disease control is vital for producers' profit and for consumer's welfare. This study aimed at investigating the immune response to Staphylococcus epidermidis intramammary infection. METHODS: A total of 14 S. epidermidis isolates from milk collected from ewes with subclinical mastitis were used. Protein extracts were prepared and analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Immunoblotting assay was performed for the detection of specific IgG and IgA in blood and milk from S. epidermidis mastitic ewes and from healthy animals. RESULTS: The presence of pathogen-specific IgG was detected in blood of both infected and healthy animals. However, in milk, pathogen-specific IgG was only identified in infected animals, while IgA was found in both groups. Proteins with 59 and 43 kDa were recognized by all immunoglobulins screened in blood and milk provided by both healthy and mastitic ewes. In addition, in milk, IgG and IgA for proteins with 35 kDa were also detected. CONCLUSION: The results have lead to propose a theory for immunoglobulin dynamics in mammary gland's defence: blood IgG1, specifically targeting intestinal antigens, is transported to the mammary gland with the main purpose of protecting the newborn, while IgG2 is specific for mammary pathogens and is transported to the mammary gland exclusively during inflammation. This study suggests that only local immunization should trigger IgG-producing cells in the mammary gland as a response to mastitis antigens. Moreover, IgA seems to be of crucial value for the defence of the ewe mammary gland, and stimulation strategies towards an increase in IgA should be addressed for mastitis prevention.
ABSTRACT
BACKGROUND CONTEXT: Percutaneous vertebroplasty (PVP) is a minimally invasive surgical procedure and is frequently performed in humans who need surgical treatment of vertebral fractures. PVP involves cement injection into the vertebral body, thereby providing rapid and significant pain relief. PURPOSE: The testing of novel biomaterials depends on suitable animal models. The aim of this study was to develop a reproducible and safe model of PVP in sheep. STUDY DESIGN: This study used ex vivo and in vivo large animal model study (Merino sheep). METHODS: Ex vivo vertebroplasty was performed through a bilateral modified parapedicular access in 24 ovine lumbar hemivertebrae, divided into four groups (n=6). Cerament (Bone Support, Lund, Sweden) was the control material. In the experimental group, a novel composite was tested-Spine-Ghost-which consisted of an alpha-calcium sulfate matrix enriched with micrometric particles of mesoporous bioactive glass. All vertebrae were assessed by micro-computed tomography (micro-CT) and underwent mechanical testing. For the in vivo study, 16 sheep were randomly allocated into control and experimental groups (n=8), and underwent PVP using the same bone cements. All vertebrae were assessed postmortem by micro-CT, histology, and reverse transcription-polymerase chain reaction (rt-PCR). This work has been supported by the European Commission under the 7th Framework Programme for collaborative projects (600,000-650,000 USD). RESULTS: In the ex vivo model, the average defect volume was 1,275.46±219.29 mm3. Adequate defect filling with cement was observed. No mechanical failure was observed under loads which were higher than physiological. In the in vivo study, cardiorespiratory distress was observed in two animals, and one sheep presented mild neurologic deficits in the hind limbs before recovering. CONCLUSIONS: The model of PVP is considered suitable for preclinical in vivo studies, mimicking clinical application. All sheep recovered and completed a 6-month implantation period. There was no evidence of cement leakage into the vertebral foramen in the postmortem examination.
