Diversity and Resistance Profiles of Human Non-typhoidal Salmonella spp. in Greece, 2003-2020.

Salmonella spp. is one of the most common foodborne pathogens in humans. Here, we summarize the laboratory surveillance data of human non-typhoidal salmonellosis in Greece for 2003-2020. The total number of samples declined over the study period (p < 0.001). Of the 193 identified serotypes, S. Enteritidis was the most common (52.8%), followed by S. Typhimurium (11.5%), monophasic S. Typhimurium 1,4,[5],12:i:- (4.4%), S. Bovismorbificans (3.4%) and S. Oranienburg (2.4%). The isolation rate of S. Enteritidis declined (p < 0.001), followed by an increase of the less common serotypes. Monophasic S. Typhimurium has been among the five most frequently identified serotypes every year since it was first identified in 2007. Overall, Salmonella isolates were resistant to penicillins (11%); aminoglycosides (15%); tetracyclines (12%); miscellaneous agents (sulphonamides, trimethoprim, chloramphenicol and streptomycin) (12%) and third-generation cephalosporins (2%). No isolate was resistant to carbapenems. In total, 2070 isolates (24%) were resistant to one or two antimicrobial classes and 903 (10%) to three and more. Out of the 1166 isolates resistant to fluoroquinolones (13%), 845 (72%) were S. Enteritidis. S. Enteritidis was also the most frequently identified serotype with a resistance to third-generation cephalosporins (37%, 62/166), followed by S. Typhimurium (12%, 20/166). MDR was most frequently identified for S. Typhimurium and its monophasic variant (resistant phenotype of ampicillin, streptomycin, tetracycline and sulphamethoxazole with or without chloramphenicol or trimethoprim).

Surveillance data for human leishmaniasis indicate the need for a sustainable action plan for its management and control, Greece, 2004 to 2018.

BackgroundThe World Health Organization (WHO) lists human leishmaniasis as a neglected tropical disease; it is not under surveillance at European level.AimWe present surveillance data for visceral (VL) and cutaneous (CL) leishmaniasis for the period 2004 to 2018 in Greece to assess their public health importance.MethodsWe extracted data from the mandatory notification system to analyse separately imported and domestic cases of VL and CL. A case was defined by clinical manifestations compatible with VL or CL and laboratory confirmation.ResultsBetween 2004 and 2018, 881 VL (862 domestic, 19 imported) and 58 CL cases (24 domestic, 34 imported) were recorded. The mean annual notification rate of domestic VL was 0.5 per 100,000 (range: 0.12-1.43/100,000) with a statistically significant increasing trend (p = 0.013). Cases were reported by all regions. The highest notification rate occurred in the age group 0-4 years (1.3/100,000). Overall 24% (164/680) of the cases were immunocompromised and their proportion increased after 2010 (p < 0.001). The mean annual notification rate of domestic CL was 0.05 per 100,000 (range: 0.01-0.19/100,000) with the highest rate in the age group 5-14 years (0.03/100,000). Cases were recorded in six of the 13 regions. Among 34 imported CL cases, 29 were foreign nationals.ConclusionVL is endemic in Greece, with an increasing trend and a considerable burden of severe disease and young children being most affected. CL is rarely reported. A sustainable action plan is needed to reduce the burden of VL and prevent local transmission of CL.

The Rising Burden of Salmonellosis Caused by Monophasic Salmonella Typhimurium (1,4,[5],12:i:-) in Greece and New Food Vehicles.

Monophasic Salmonella typhimurium is of increasing importance worldwide. Here we present the available data regarding monophasic S. typhimurium from 2007 to 2019 in Greece, in order to assess its public health impact. Surveillance data, data on antimicrobial resistance, molecular typing by pulsed-field gel electrophoresis (PFGE), and results of the investigation of monophasic S. typhimurium outbreaks were analyzed. Overall, 403 cases were identified; 329 (81.6%) sporadic and 74 (18.4%) related to two community outbreaks in 2017. A total of 305 isolates from sporadic cases tested for antimicrobial resistance revealed resistance to ampicillin, streptomycin, sulphamethoxazole, and tetracycline (41.3%). Some 23.3% were further resistant to trimethoprim and 5.2% were also resistant to chloramphenicol. Outbreak 1 in 2017 with 37 identified cases was attributed to the consumption of raw milk from a vending machine and isolates were resistant to ampicillin, streptomycin, sulphamethoxazole, tetracycline, and trimethoprim. Outbreak 2 also with 37 cases was attributed to the consumption of pork and isolates were resistant to the five above mentioned antibiotics plus chloramphenicol. The number of human monophasic S. typhimurium isolates is low; however, since 2009, it has been among the five most frequently identified serotypes in Greece. Investigation of the outbreaks revealed that other vehicles apart from pork may be implicated in the occurrence of outbreaks.

Impact of Salmonella Control Programmes in Poultry on Human Salmonellosis Burden in Greece.

Since 2008, veterinary authorities in Greece have implemented national control programmes (NSCPs) targeting S. Enteritidis (SE) and S. Typhimurium (ST) in poultry. We assessed the effect of the programs on the reported number of human isolates. Using monthly data for 2006-2017, we defined two groups (SE, ST) and one control group with serotypes unrelated to poultry or eggs. For SE we also analysed data for 2006-2015 due to a multi-county SE outbreak in 2016. We performed an interrupted time series analysis and used a negative binominal regression model. For both SE and ST, there was no significant trend of the isolation rate before or after NSCPs' introduction. After the NSCPs' introduction there was an increasing rate (IRR: 1.005, 95% CI: 1.001-1.008) for control serotypes and a decreasing one for SE (IRR: 0.990, 95% CI: 0.986-0.995) (for 2009 to 2015 analysis). From 2006 to 2017, NSCPs had a statistically significant impact on the number of SE isolates that decreased by 49% (IRR:0.511, 95% CI: 0.353-0.739). No impact was shown on the number of ST (p-value = 0.741) and control isolates (p = 0.069). As a conclusion, NSCP's implementation was associated with decreased SE isolates and overall burden of salmonellosis; however further measures aiming at human salmonellosis due to ST, should be considered.

Antibody Response Following Pre-Exposure Immunization Against Rabies in High-Risk Professionals.

Vaccination against rabies and routine antibody testing of subjects participating in programs for the surveillance and control of rabies in animals is strongly recommended. The scope of this study is to describe the antibody level as measured by a commercial enzyme-linked immunosorbent assay (ELISA) after primary and booster intramuscular vaccination with a purified vero-cell rabies vaccine (PVRV) in high-risk professionals and to determine the influence of an array of factors on antibody level, that is, time elapsed since primary immunization series and booster dose, sex, age, pathologic conditions, high-risk occupation, and peak antibody level after initial scheme and booster dose. A primary series of three doses of PVRV was administered and a commercial ELISA was recommended 14 days postimmunization with continuous repetition at 6 months and yearly intervals for the laboratory personnel and the rest of the professionals, respectively. The protective antibody titer was defined as a minimum of 0.5 equivalent units/mL (EU/mL) (seroconvertion) and a booster dose was applied if the titer was determined nonprotective. The seroconversion rate (SCR) after primary vaccination was 100%, with a geometric mean titer (GMT) of 2.90 EU/mL (interquartile range [IQR]: 1.85-3.45). After booster vaccination due to nonprotective titer, the SCR was 100% and the GMT increased by 678% (95% confidence interval [CI]: 514-887) reaching 4.25 EU/mL (IQR: 4.00-4.60), 2.5 times higher than the GMT elicited by the primary vaccine scheme in the respective recipients. The titer dropped by 1.20% per month (95% CI: 0.52-1.89) regardless of booster administration or any other factor. Women had 51% higher titer compared with men (95% CI: 6-116). High-risk professionals should be verified for adequate antibody titers, but routine administration of a single booster dose of PVRV 1 year after the primary series could be considered; more evidence is needed to support the benefit in terms of immunity and logistics.

Surveillance of Rabies Postexposure Prophylaxis in Greece: 4 Years Experience.

Rabies reemerged in Greek fauna during October 2012, 25 years after the last report in animals and 42 after the last human case. This study examined the administration of rabies postexposure prophylaxis (PEP) in humans over the period 2012-2016. A total of 1,454 individuals (62.6% males) received PEP. The vast majority (92.3%) of regimens was initiated in high risk for animal rabies areas (11.1 PEP per 100,000 residents per year). Among the exposed, 77.0% presented at a healthcare setting during the first 24 h after the incident; more severe injuries were associated with faster presentation (p < 0.05). A vaccine series was administered in 54.5% of exposed persons whereas 43.7% received both vaccine and immunoglobulin and 1.7% immunoglobulin only. Exposure to stray dogs represented 68.4% of all incidents. In exposures occurring in nonhigh risk for rabies areas, bat was the second most frequently involved animal (13.4% of incidents). All dogs and cats evaluated by a veterinarian and laboratory-confirmed rabid were initially deemed suspect for rabies during clinical examination or according to history. No human rabies cases were recorded during the period of the study. Surveillance of PEP represents a valuable tool for aiding assessment of present and future demands in prophylaxis biologicals, outlining the epidemiological profile of exposures and planning effective policies for the management of exposure incidents.

Efficacy of oral rabies vaccination in individual age groups of juvenile red foxes.

Although juvenile red foxes (Vulpes vulpes) are considered a single age group, essential for monitoring the effectiveness of the oral rabies vaccination (ORV), there appear to be significant differences among age subgroups. Herein, a subset of 335 foxes aged 0-1 year that had not consumed bait in previous campaign were collected for monitoring the effectiveness of the first seven ORV campaigns in Greece, carried out from 2013 to 2017. These juveniles were additionally assigned to three individual 4-month age groups, according to the exact date on which they were killed. The aim was to identify differences in seroconversion rate and bait uptake level and determine whether reconsideration is needed in the way that ORV monitoring is being implemented and evaluated. Statistically significant differences were observed following the analysis of mandible bone, teeth and blood samples obtained from 1-4 and 5-8-month old foxes as compared to the respective samples derived from 9-12-month old animals, whereas no differences were revealed in samples between foxes aged 1-4 and 5-8 months. Hunting juveniles during the whole period of spring ORV campaigns monitoring should be reevaluated and even discouraged. On the contrary, juvenile foxes hunted for the evaluation of autumn campaigns, aged > 8 months, had similar assessment rates to adult individuals and are equally helpful for assessing the efficacy of an ORV campaign. Taking the above into consideration and by distinguishing recent and old tetracycline uptake, ORV monitoring and evaluation could be performed in an alternative, more comprehensive way.

First four Oral Rabies Vaccination campaigns of the red foxes in Greece: Evaluating factors and assessment.

Following the last animal rabies outbreak in Greece in 2012, Oral Rabies Vaccination (ORV) campaigns of red foxes (Vulpes vulpes) were conducted in order to halt the spread of the disease, as widely and effectively have also been implemented in other countries. The present study aims to report the main outcomes following the first Greek ORV campaigns during autumn 2013, 2014, 2015 and spring 2016, to assess their effectiveness and to investigate factors potentially related to their success. Blood samples, mandible bones and teeth, derived by 452 foxes, were tested for rabies antibody titration, animal age determination and tetracycline (TTC) detection. The laboratory results obtained were statistically analyzed. High seroprevalence and TTC detection rates were obtained following the autumn campaigns studied, while these rates were significantly reduced following the spring campaign. The year or the season of the vaccination campaign, the estimated age group of the animal and the geographical Regional Unit (RU), where the animal was hunted, were identified as important factors. On the contrary, no significance could be ascertained for TTC detection based on exclusively previous uptake, use of filter paper, blood sample type and quality, as well as sex of animal. Based on the monitoring results achieved, the first ORV campaigns conducted in the country can be generally considered to be satisfactory. No positives cases were detected since May 2014. Seasonal, geographical parameters and factors related to fox ecology may interfere with monitoring results and should be always considered when planning future ORV programs.

Evaluation of the first oral rabies vaccination campaign of the red foxes in Greece.

Following the late 2012 recurrence of rabies in wild foxes (Vulpes vulpes) in central and north-western Greece, the first oral fox vaccination campaign co-financed by the European Union (EU) and the Greek state budget, was implemented. Initially, it involved 24 regional units of the Greek territory during the period October-December 2013. Vaccine-baits were aerially distributed by fixed-wing aircrafts. Vaccines were scattered along parallel flight paths 500m apart in order to optimize aerial missions and achieve homogeneous distribution. A geographical information system was used to objectively evaluate bait distribution. This system identified areas of inadequate bait density that would require additional flights. A total number of 1,504,821 baits were distributed covering an area of 54,584.29km(2). To assess the effectiveness of oral vaccination campaign a monitoring program was introduced, which entailed examination of serum samples and canine teeth derived from red foxes collected in the field. The laboratory analysis revealed 60% seropositivity and detection of tetracycline biomarker in 70% of the foxes tested.

Rabies in Greece; historical perspectives in view of the current re-emergence in wild and domestic animals.

Greece has been rabies free since 1987 while no human cases have been seen since 1970. The re-emergence of rabies in Northern Greece during 2012-2013 in wild and domestic animals prompted a systematic review of historical evidence of the presence of the disease in the country from ancient years till the present. Historical data is presented along with efforts to prevent disease in animals and humans especially during the high prevalent periods in the country in the mid-20th century. These efforts serve as a guide to current extensive efforts to prevent spread especially in the wild and domestic animal populations.