Predictive statistical models for monitoring antimicrobial resistance spread in the environment using Apis mellifera (L. 1758) colonies.

The rise of antimicrobial resistance (AMR) is one of the most relevant problems for human and animal health. According to One Health Approach, it is important to regulate the use of antimicrobials and monitor the spread of AMR in the environment as well. Apis mellifera (L. 1758) colonies were used as bioindicators thanks to their physical and behavioural characteristics. During their foraging flights, bees can intercept small particles, including atmospheric particulate matter, etc., and also microorganisms. To date, the antimicrobial surveillance network is limited to the sanitary level but lacks into environmental context. This study aimed to evaluate the use of A. mellifera colonies distributed throughout the Emilia-Romagna region (Italy) as indicators of environmental antimicrobial-resistant bacteria. This was performed by creating a statistical predictive model that establishes correlations between environmental characteristics and the likelihood of isolating specific bacterial genera and antimicrobial-resistant strains. A total of 608 strains were isolated and tested for susceptibility to 19 different antimicrobials. Aztreonam-resistant strains were significantly related to environments with sanitary structures, agricultural areas and wetlands, while urban areas present a higher probability of trimethoprim/sulfamethoxazole-resistant strains isolation. Concerning genera, environments with sanitary structures and wetlands are significantly related to the genera Proteus spp., while the Escherichia spp. strains can be probably isolated in industrial environments. The obtained models showed maximum values of Models Accuracy and robustness (R2) of 55 % and 24 %, respectively. The results indicate the efficacy of utilizing A. mellifera colonies as valuable bioindicators for estimating the prevalence of AMR in environmentally disseminated bacteria. This survey can be considered a good basis for the development of further studies focused on monitoring both sanitary and animal pathology, creating a specific network in the environments of interest.

Using honey bee colonies to monitor phenotypic and genotypic resistance to colistin.

Colistin is a polymyxin antimicrobic mainly used to treat infection caused by multi-drug resistant Gram-negative bacteria. Mechanisms of colistin resistance are linked to the mobile colistin resistance (mcr) genes, which are transferable within mobile plasmids. Currently, there is limited research on the environmental dissemination of these genes. The behavioural and morphological characteristics of Apis mellifera L. make honey bees effective environmental bioindicators for assessing the prevalence of antimicrobial-resistant bacteria. This study aims to evaluate the colistin phenotypic and genotypic resistance in environmental Gram-negative bacteria isolated from foraging honey bees, across a network of 33 colonies distributed across the Emilia-Romagna region in Italy. Phenotypic resistances were determined through a microdilution assay using the minimum inhibitory concentration (MIC) with dilutions ranging from 0.5 µg/ml to 256 µg/ml. Strains with MIC values gather than 2 µg/ml were classified as resistant. Also, the identification of the nine mcr genes was carried out using two separate multiplex PCR assays. The study found that 68.5% of isolates were resistant and the genus with the higher resistance rates observed in Enterobacter spp. (84.5%). At least one mcr gene was found in 137 strains (53.3%). The most detected gene was mcr5 (35.3%), which was the most frequently detected gene in the seven provinces, while the least observed was mcr4 (4.8%), detected only in two provinces. These results suggested the feasibility of detecting specific colistin resistance genes in environmentally spread bacteria and understanding their distribution at the environmental level, despite their restricted clinical use. In a One-Health approach, this capability enables valuable environmental monitoring, considering the significant role of colistin in the context of public health.

Active surveillance of antimicrobial resistance in companion animals: A pilot study in a Spanish Veterinary Teaching Hospital.

The role of small animal veterinary hospitals in the onset and dissemination of antimicrobial-resistant organisms (AMROs) is still not clear, and the implementation of an internal surveillance systems is a cost-effective tool to better understand their impact. The aim of this study was to describe a pilot program of active surveillance in a Spanish Veterinary Teaching Hospital, developed to estimate the detection frequency of AMROs in the commensal flora of patients and in the environment. Surveillance was focused on Methicillin-resistant Staphylococci (MRS), third generation cephalosporins resistant gram-negative bacteria (3GCR-GNB), and carbapenems-resistant gram-negative bacteria (CR-GNB). Oral and perirectal swabs were collected in the same dogs and cats hospitalized > 48 h, at their admission and before their discharge. Out of 50 patients sampled, 24% (12/50) were carriers at admission of at least one of the three investigated AMROs. Twenty-eight percent of patients (14/50) acquired at least one AMRO during the hospital stay. MRS detection frequency at admission was 12% (6/50), while acquisition was 6% (3/50). 3GCR-GNB detection frequency was 14% at admission (7/50) and acquisition 22% (11/50), while CR-GNB detection frequency was 2% at admission (1/50) and acquisition 2% (1/50). Environmental surveillance (98 samples) showed a total detection frequency of 22.4% for MRS (22/98), 2% for 3GCR-GNB and CR-GNB (2/98). Clinical staff' shoe soles showed high detection frequency for MRS (50%). 3GCR Escherichia coli was the most isolated species in patients (n = 17). The results show how active surveillance can be used as a tool to assess the impact of AMROs in veterinary hospitals to subsequently build up tailored control plans based on specific issues.

Case report: First isolation of Yersinia pseudotuberculosis from the blood of a cat.

A 14-year-old female domestic short-haired cat with a diagnosed diabetes mellitus and acromegaly was presented for lethargy and dysorexia. On clinical presentation, the patient showed hyperglycemia, hyperthermia, dull mentation, and dehydration. With the suspicion of an inflammatory or infectious complication of diabetes, she was hospitalized with constant rate infusion of insulin, and empirical ampicillin sulbactam was started. Blood culture revealed positivity for Yersinia pseudotuberculosis and the septic picture was confirmed by blood analysis, with leukocytosis, neutrophilia, and an increased serum amyloid A concentration. The isolated Y. pseudotuberculosis strain showed susceptibility to every antimicrobial tested. During the second day of hospitalization, the onset of hypoglycemia and hypotension was treated with norepinephrine and glucose in fluid therapy. The cat recovered well and was discharged with insulin and amoxicillin-clavulanate. This is the first case of septicemia associated with Y. pseudotuberculosis in a cat, suspected of developing the infection after contact with natural reservoirs such as rodents or birds. This route of transmission should be highlighted especially in relation to the zoonotic potential of the bacteria.

Avaliação dos níveis de hemoglobina glicada em pacientes com nefropatia diabética / Assessment of glycated hemoglobin rates in patients with diabetic nephropathy

A nefropatia diabética (ND) é uma complicação comum do diabetes mellitus (DM), sendo que a hiperglicemia está associada ao desenvolvimento das lesões glomerulares observadas na ND. Neste estudo foram avaliados os níveis de HbA1c em pacientes diabéticos com e sem nefropatia. Foram determinados os níveis de glicose, colesterol total, HDL-colesterol, LDL-colesterol, triglicérides, creatinina, taxa de filtração glomerular (TFG), HbA1c e microalbuminúria em pacientes diabéticos tipo 2 sem nefropatia (n=50) e com nefropatia (n=57). Os valores de HbA1c nos pacientes dos grupos sem nefropatia e com nefropatia foram, respectivamente, de 6,4 ± 0,1% e 7,0 ± 0,1%. O grupo de pacientes com nefropatia apresentou níveis mais elevados de microalbuminúria do que o grupo sem nefropatia (62,7 ± 7,3 mg/L versus 7,4± 0,7 mg/L, P<0,001), e a TFG obtida no grupo com nefropatia foi significativamente menor do que a observada no grupo sem nefropatia (67,9 ± 2,5 mL/min/1,73 m2 77,4 ± 2,3 mL/min/1,73 m2, 0,01). Não foram observadas diferenças significativas entre os grupos quanto aos níveis de glicose e perfil lipídico. Os valores de hemoglobina glicada foram mais elevados nos pacientes com nefropatia, o que evidencia o papel deste marcador no controle do diabetes e na prevenção de risco para o desenvolvimento das complicações crônicas da doença.

Diabetic nephropathy (DN) is a common renal disease associated with diabetes mellitus (DM), and hyperglycemia is associated with the development of glomerular lesions. This study evaluated the rate of HbA1c in patients with nephropathy and in those without nephropathy. Glucose, total cholesterol, HDLcholesterol, LDL-cholesterol, triglycerides, creatinine, glomerular filtration rate (GFR), HbA1c and urinaryalbumin contents were determined in 50 diabetic type 2 patients without nephropathy and in 57 patients with nephropathy. The HbA1c in patients without and with nephropathy were 6.4 ± 0.1% and 7.0 ± 0.1%, respectively. In addition, the urinary albumin contents were higher in ND group (62.7 ± 7.3 mg/L versus 7.4 ± 0.7 mg/L, P<0.001), while GFR values were lower in ND patients than in patients without nephropathy (67.9 ± 2.5 mL/min/1.73 m2 versus 77.4 ± 2.3 mL/min/1.73 m2, P<0.01). No significant differences were observed for glucose and lipid profile. The highest rates of HbA1c were detected in patients with DN, and these findings support the use of this biomarker for diabetes control and for monitoring the patients and to prevent them from getting diabetes-associated diseases.

Intoxicação em cães e gatos: diagnóstico toxicológico empregando cromatografia em camada delgada e cromatografia líquida de alta pressão com detecção ultravioleta em amostras estomacais / Intoxication in dogs and cats: toxicological diagnosis using thin layer chromatography and high pressure liquid chromatography with ultraviolet detection in stomach samples

Agrotóxicos e raticidas são responsáveis por inúmeras intoxicações humanas e animais. Dados preliminares sugerem que o uso ilegal desses compostos com a finalidade de intoxicação fatal em pequenos animais é uma prática comum na região central do Estado do Rio Grande do Sul. O Laboratório de Toxicologia (LATOX) recebe amostras de casos em que a principal suspeita é a intoxicação por agrotóxicos ou raticidas (lícitos e ilícitos). O presente estudo teve como objetivo apresentar um levantamento das intoxicações de pequenos animais, analisadas pelo LATOX no período de 2004 a 2008, sendo identificados os xenobióticos por meio de métodos analíticos otimizados pelo laboratório, incluindo screening por cromatografia em camada delgada (CCD) e possível confirmação por cromatografia líquida de alta eficiência (CLAE-UV). No período, foram analisadas 68 amostras oriundas de intoxicações em cães e gatos. As amostras biológicas analisadas foram o estômago e o conteúdo estomacal, das quais a CCD permitiu a identificação de carbamatos, warfarina e estricnina. Esta mostrou ser uma técnica qualitativa eficiente e adequada para esse propósito, além de ser relativamente rápida, de baixo custo e de sofrer pouca interferência de componentes da matriz. Também foi realizado um screening toxicológico por CLAE-UV para os carbamatos n-metilados: aldicarb, carbaril, carbofuran e propoxur. O resultado das análises indicou que o principal agente tóxico encontrado foi o aldicarb (chumbinho), responsável por 39,7 por cento das intoxicações (27 casos), seguido por estricnina (seis casos), warfarina (três casos) e monofluoracetato de sódio (um caso). Sendo assim, o "chumbinho" foi o principal agente envolvido em intoxicações de cães e gatos na região central do Estado no período avaliado, e os métodos analíticos CCD e CLAE-UV podem ser utilizados de forma eficiente na rotina laboratorial para identificação e confirmação dos xenobióticos mais envolvidos nessas intoxicações.

The pesticides and rodenticides are responsible for several human and animal intoxications. Preliminary data suggest that the illegal use of these compounds to poison small animals is a common practice in the central region of the Rio Grande do Sul state. The Laboratory of Toxicology received samples, of cases in which the main suspected cause of death is poisoning by pesticides or exogenous rodenticides (licit and illicit). The aim of this study was show the survey of small animals poisoned and analyzed by LATOX during the period of 2004 to 2008 identifying the xenobiotics by optimized analytical methods, including screening by thin-layer chromatography (TLC) and a possible confirmation by high performance liquid chromatography - ultraviolet detection (HPLC-UV). In this period 68 samples were analyzed from small poisoned animals. The biological samples analyzed were stomach and stomach content and the TLC permitted carbamates, warfarin and stricnine identification. This proved to be an efficient and adequate technique for this purpose, relatively fast, inexpensive and with low matrix interference. The screening by HPLC for N-methyl carbamates was also realized: aldicarb, carbaryl, carbofuran and propoxur. The analysis showed that the main toxic agent found was aldicarb (chumbinho), responsible for 39.7 percent of poisoning (27 cases), followed by stricnine (6 cases), warfarin (3 cases) and sodium monofluoracetate (1 case). Thus, the 'chumbinho' was the main agent involved in dogs and cats poisoning in the central region of the state, during the analyzed period. The analytical methods TLC and HPLC can be used efficiently in laboratorial routine for identification and confirmation of xenobiotics involved in these poisonings.