An Atlas of Surra in Spain: A Tool to Support Epidemiological Investigations and Disease Control.

Trypanosomosis is a global animal issue, causing significant economic losses, particularly in Africa. In Spain, only one pathogenic species, Trypanosoma evansi, has been identified so far. It was first detected in a dromedary camel in the Canary Islands in 1997. Since then, numerous cases of the disease, known as Surra, have been diagnosed, prompting various studies and efforts in control and surveillance. Given the lack of a comprehensive database that consolidates the most relevant data in this area, the development of a national atlas, with a focus on the Canary Islands, to incorporate all available information on T. evansi in Spain became a necessity. For the development of the atlas, a repository was constructed, encompassing a range of datasets and documents spanning from 1997 to 2022. Information from each source, and in particular georeferenced locations and results of blood tests on animals, were extracted and integrated into a comprehensive database. A total of 31 sources were analysed, providing a total of 99 georeferenced locations and 12,433 animal samples. Out of these samples, 601 (mostly from dromedaries) were found to be positive for T. evansi. The Card Agglutination Test for T. evansi (CATT/T. evansi), a serological test, was the most commonly used diagnostic method, and it showed a higher prevalence for all tested animal species. Positive cases were mainly concentrated in the Canary Islands, specifically in the eastern islands, with isolated cases found in the province of Alicante (Iberian Peninsula). This atlas provides an overview of the history and occurrence of Surra in Spain, and it represents a valuable tool for future control initiatives and for research. Still, the need for more studies remains, especially for further testing of potential hosts other than camelids and for the examination of their potential transmission vectors.

Prevalence, antimicrobial resistance, and genetic lineages of nasal Staphylococcus aureus among medical students at a Spanish University: detection of the MSSA-CC398-IEC-type-C subclade.

Medical students could be a potential source of Staphylococcus aureus transmission to patients. This cross-sectional study involved samples collected from both nasal nostrils. Samples were processed for S. aureus recovery; the antimicrobial resistance (AMR) phenotype was determined by disc diffusion assays and the spa types and AMR genotypes by PCR/sequencing. A structured questionnaire was administered to students to collate data related to potential risk factors of nasal colonization. Ninety-eight students were included, 50 % were colonized by S. aureus and 12.2 % by MRSA. The mecA gene was detected in all MRSA isolates. The MSSA-CC398-IEC-type C lineage was found among 16.3 % of nasal carriers, of which t571 was the predominant spa-type. MRSA isolates were ascribed to spa types t2226 (CC5, 12 isolates) and t3444 (new spa type, 1 isolate). All MRSA were multi-drug resistant and MSSA were predominantly resistant to erythromycin-clindamycin (inducible-type, mediated by ermT gene). High rates of S. aureus and MRSA nasal carriages were observed in this study. The predominance of the CC398 lineage among MSSA (emergent invasive lineage) represent a relevant finding of public health concern. The role of medical students as potential source of MRSA and MSSA-CC398 transmissions in hospital and community needs to be elucidated in detail.

Surveillance and control of Trypanosoma evansi in the canary Islands: A descriptive analysis.

This study examines the occurrence of Surra, a disease caused by Trypanosoma evansi, in camels in the Canary Islands. The 1997 detection of T. evansi in camels in the Canary Islands led to the implementation of an initial control program, resulting in a decrease in prevalence. Following an outbreak in 2014, and due to the impossibility of eradicating it using the conventional measures, a lazaret was set up to separate positive and suspicious animals, in addition to the control measures previously implemented. Stomoxys calcitrans was the only vector captured, and no other animals tested were found to be positive for T. evansi. In November 2019, the last camels that tested serologically positive were detected; however, since February 2018, no camels that tested positive for PCR have been found in the farms were the outbreak was detected, suggesting that the sanitary measures implemented are adequate. The duration of the outbreak control and potential eradication for the disease has yet to be established. This study provides evidence to facilitate the control of African Animal Trypanosomosis in endemic areas of the world, which may contribute to revise the World Organization for Animal Health (WOAH) protocol to implement recommendations of surveillance and control strategies for animal Trypanosomosis in camels.

Antimicrobial resistance patterns of bacteria isolated from chicks of Canarian Egyptian vultures (Neophron percnopterus majorensis): A "one health" problem?

Antimicrobial resistance in Gram-negative bacteria isolated from cloacal samples of chicks of Canarian Egyptian vultures was investigated. Prevalence of Salmonella was also studied. Forty-seven isolates, obtained from 23 animals, were analysed. Escherichia coli (n = 29), Proteus mirabilis (n = 17) and Salmonella spp. (n = 1) were identified using API 20E system. Antimicrobial susceptibility to 13 antibiotics included in nine different categories was determined using disk-diffusion technique. The higher percentages of susceptible E. coli isolates were found for aminoglycosides and cefoxitin, and the lower ones were found for ampicillin, enrofloxacin and tetracycline. Proteus mirabilis isolates were susceptible to most of antimicrobials tested. Multidrug resistance patterns were found in 13 E. coli and four Proteus mirabilis. Salmonella spp. was detected in one chick (4.37%), and the isolate was also resistant to ampicillin and tetracycline. Thirteen E. coli isolates and four Proteus isolates showed a multidrug-resistance pattern, being resistant at least to one antibiotic in three or more different antimicrobial categories. This high level of antibiotic resistance in chickens of an endangered bird may be a limitation for possible treatments of infections in this species, as well as representing a source of resistant bacteria for animal care staff and for other animals in wildlife recovery centres. A "One Health" approach to this problem is necessary to reduce the levels of antimicrobial resistance in wild birds.

Staphylococcus aureus and Methicillin-Resistant Coagulase-Negative Staphylococci in Nostrils and Buccal Mucosa of Healthy Camels Used for Recreational Purposes.

Several different species of animals host staphylococci as normal microbiota. These animals can be a source of staphylococci zoonotic infections. People with routine or occupational exposure to infected/colonized animals are at risk of a potential transmission. Therefore, we aimed to investigate the presence of S. aureus and other staphylococci in camels used for recreational purposes as well as their antimicrobial resistance, virulence factors and genetic lineages. A total of 172 samples were collected from 86 healthy camels (nose and mouth) from different farms located in the Canary Islands, Spain. Antimicrobial susceptibility testing was performed against 14 antimicrobial agents. The presence of virulence genes was studied by PCR. Multilocus sequence typing, spa typing and agr typing were performed in all S. aureus isolates. From the 86 camels tested, 42 staphylococci were isolated, of which there were 11 S. aureus, 13 S. lentus, 12 S. sciuri, 3 S. xylosus, S. epidermidis, S. hominis and S. chromogenes. Staphylococci isolates were resistant to penicillin, ciprofloxacin, clindamycin and fusidic acid. All S. aureus isolates harbored the hla, hlb and hld virulence genes. S. aureus isolates were ascribed to three sequence types (STs) and three spa types. All S. aureus isolates belonged to agr type III. Camels from Gran Canaria used in recreational purposes have a moderate prevalence of S. aureus and other coagulase-negative staphylococci. Nevertheless, S. aureus isolates are susceptible to almost all antibiotics tested.

Microbiological aspects of osteomyelitis in veterinary medicine: drawing parallels to the infection in human medicine.

Osteomyelitis is a challenging infectious disease affecting humans and animals. It is difficult to diagnose because, in many cases, symptoms are non-specific and, for example in implant-related cases, can appear long time after surgery. In addition to this, it is also difficult to treat due to the need to find the appropriate antibiotic regime and delivery system to reach the site of infection and to avoid development of bacterial resistance. The central purpose of this review is to compare the microbiological aspects of osteomyelitis in human and veterinary medicine, with the aim of improving the microbiological diagnosis and treatment of this infection in animals. Furthermore, the study of osteomyelitis in animals may help to improve the development of animal models for testing new treatments in humans. Host factors and underlying conditions have been studied mainly in humans, although aspects as immunodeficiency have been described in some veterinary cases. Even when Staphylococcus aureus is still considered the most prevalent causing microorganism, this prevalence should be reviewed using molecular diagnostic techniques, and this could affect treatment options. New approaches to treatment include local delivery of antibiotics using different biomaterials, antimicrobial photodynamic therapy, and new antimicrobial compounds. We would like to remark the need of large, high-quality clinical trials and of the development of guides for the diagnosis and treatment of osteomyelitis in different animal species.

Multidrug-Resistant Phenotypes of Escherichia coli Isolates in Wild Canarian Egyptian Vultures (Neophron percnopterus majorensis).

The presence of multidrug-resistant (MDR) Escherichia coli in cloacal samples from Canarian Egyptian vultures was investigated. Samples were obtained from chicks (n = 65) and from adults and immature birds (n = 38). Antimicrobial susceptibility to 16 antibiotics included in 12 different categories was determined for 103 E. coli isolates. MDR was defined as acquired non-susceptibility to at least one agent in three or more antimicrobial categories. Forty-seven different resistance phenotypes were detected: 31 MDR (41 isolates) and 16 non-MDR (62 isolates). One isolate was resistant to all 12 antimicrobial categories and 2 phenotypes included resistance to 9 antimicrobial categories. Imipenem resistance was included in five MDR phenotypes, corresponding to five different isolates. Statistically significant differences in prevalence of MDR-phenotypes were found between chicks in nests and the rest of the animals, probably due to the shorter exposure time of chicks to antimicrobials. The main risk derived from MDR bacteria in scavengers is that it threatens the treatment of wild animals in rescue centres and could be transferred to other animals in the facilities. In addition to this, it could pose a health risk to veterinarians or other staff involved in wildlife protection programmes.

Molecular diversity of Extended-spectrum ß-lactamase-producing Escherichia coli from vultures in Canary Islands.

Antimicrobial resistance among isolates from wild animals is increasingly reported. Extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae, and particularly Escherichia coli, have spread worldwide as one of the most common multidrug-resistant organisms. The aim of this study was to determine the carriage rate of ESBL-producing E. coli isolates and their genetic characteristics in wild vultures from the Canary Islands. Faecal samples were collected from 22 apparently healthy free-ranging (wild) vulture chicks from Lanzarote and Fuerteventura (Canary Islands) during July 2019. They were seeded in MacConkey agar supplemented with cefotaxime (2 µg ml-1 ). Colonies with typical morphology of E. coli were identified by MALDI-TOF-MS. Antimicrobial susceptibility was done by disk diffusion. Phenotypic detection of ESBL was performed by double-disk tests. The presence of blaCTX-M , blaSHV , blaTEM , blaKPC and blaOXA-48 genes, as well as mcr-1 (colistin resistance), tetA/tetB and int1 gene, was tested by PCR/sequencing. Phylogenetic groups and multilocus sequence typing (MLST) were determined by PCR/sequencing. ESBL-producing E. coli isolates were detected in 5/22 tested animals (22.7%), and all isolates (one/animal) carried blaCTX-M genes: blaCTX-M-15 (n = 3) and blaCTX-M-55 (n = 2). ESBL-positive isolates were ascribed to phylogenetic group D (two isolates), B1 (two isolates) and A (one isolate), and five sequence types were detected (ST/phylogenetic-group/ESBL): ST515/B1/CTX-M-15, ST1290/A/CTX-M-15, ST38/D/CTX-M-15, ST457/D/CTX-M-55 and ST6448/B1 /CTX-M-55; this suggests a genetic diversity among these isolates. Three CTX-M-15-producing isolates contained the blaTEM gene and one the tetA gene. To our knowledge, this appears to be the first report of ESBL-producing E. coli in vulture chicks from the Canary Islands.

Escherichia coli Producing Extended-Spectrum ß-lactamases (ESBL) from Domestic Camels in the Canary Islands: A One Health Approach.

OBJECTIVE: This work aimed to determine the carriage rate of ESBL-producing Escherichia coli as well as their genetic characteristics in camels from the Canary Islands, Spain. METHODS: Fecal samples were recovered from 58 healthy camels from Gran Canaria (n = 32) and Fuerteventura Islands (n = 26) during July 2019. They were seeded on MacConkey (MC) agar no supplemented and supplemented (MC + CTX) with cefotaxime (2 µg/mL). Antimicrobial susceptibility was determined by disk diffusion test (CLSI, 2018). The presence of blaCTX-M, blaSHV, blaTEM,blaCMY-2 and blaOXA-1/48 genes was tested by PCR/sequencing. Furthermore, the mcr-1 (colistin resistance), tetA/tetB (tetracycline resistance), int1 (integrase of class 1 integrons) and stx1,2 genes were analyzed. Phylogenetic groups and sequence types were determined by specific-PCR/sequencing for selected isolates. RESULTS: E. coli was obtained from all the 58 camels in MC media (100%) and in five of them in MC + CTX media (8.6%). Furthermore, 63.8% of E. coli isolates recovered from MC agar were susceptible to all the antibiotics tested. The five E. coli isolates recovered from MC + CTX media were characterized and two of them were ESBL-producers (3.4%). Both ESBL-producer isolates carried the blaCTX-M-15 gene and belonged to the lineages ST3018 (phylogroup A) and ST69 (phylogroup B1). The 3 ESBL-negative isolates recovered from MC-CTX plates were ascribed to phylogroup-B1. CONCLUSIONS: Camels can be a source of ESBL-producer bacteria, containing the widespread blaCTX-M-15 gene associated with the lineages ST3018 and ST69.

Microorganisms Resistant to Antimicrobials in Wild Canarian Egyptian Vultures (Neophron percnopterus majorensis).

Due to their predatory habits, raptors may serve as indicators of the presence of antimicrobial-resistant bacteria in the environment, but they also represent a public health risk for livestock and humans because they can act as reservoirs, sources and spreaders of these bacteria. Our objective was to determine the presence of antimicrobial-resistant bacteria in cloacal samples of Canarian Egyptian vultures (Neophron percnopterus majorensis), an endemic bird of prey. One hundred and forty-two cloacal swabs were obtained; Escherichia coli was isolated from 80.28% and Salmonella from 6.3% of these samples. Low levels of susceptibility to ampicillin, tetracycline and trimethoprim/sulfamethoxazole were found. About 20% of the isolates were resistant or presented intermediate susceptibility to fluoroquinolones. Surprisingly, we found isolates resistant to imipenem (6.96%). Isolates from chicks were more susceptible to antimicrobial drugs than adult and immature birds. About 50% of E. coli isolates were resistant to ampicillin, tetracycline and trimethoprim/sulfamethoxazole, and about 20% to piperacillin, enrofloxacin and marbofloxacin. High percentages of isolates of Salmonella were found to be resistant to cephalexin (88%) and aminoglycosides (greater than 77%). Our results support the idea that raptors could act as reservoirs of Salmonella and antimicrobial-resistant bacteria, posing a risk not only to wildlife but also to livestock and the human population, thus reinforcing the need to minimize the exposure of wildlife to antimicrobial agent through human and livestock waste.

Virulence factors in coagulase-positive staphylococci of veterinary interest other than Staphylococcus aureus.

Coagulase-positive Staphylococci (CoPS) can exist as commensals in humans, companion and food-producing animals, but can cause severe or even lethal diseases. Exchange of these bacteria between humans and animals has been described. Special attention has been focused on Methicillin-Resistant Staphylococcus aureus, but other CoPS can also represent an important threat. In addition to significant antimicrobial resistance, these bacteria may carry a plethora of virulence factors - molecules that allow bacteria to establish on or within a host and increase their ability to cause disease. These virulence factors have been widely described in S. aureus but information about other species of CoPS is scarce. The aim of this paper is to review the recent literature about the virulence factors of non-aureus CoPS of animal origin. Their possible effects on human health are also described. The role and prevalence of different virulence factors including leukocidins, hemolysins, adhesins, enterotoxins, exfoliative and toxic shock syndrome toxins as well as superantigen-like proteins are addressed. The effect of these virulence factors on human health is also described. The possibility of misdiagnosis of species of CoPS has been demonstrated in human clinical samples. Prevalence of zoonotic infections could be higher than thought and medical laboratories should be aware of these other staphylococcal species. In keeping with the 'One Health' approach to animal and human disease, medical professionals, veterinarians and health workers should be aware of the risks derived from exposure to these bacteria in people in close contact with animals, including pet owners, farmers and veterinarians themselves.

Preliminary evidence of the seroprevalence and risk factors associated with Chlamydia abortus infection in goats on the Canary Islands, Spain.

The aims of this cross-sectional study were to estimate the prevalence of IgG antibodies against Chlamydia abortus, the cause of enzootic abortion, in goats and to determine its associated risk factors on the Canary Islands. A total of 325 goats from 11 non-vaccinated herds were sampled and assessed using a commercial ELISA kit. Related data were also obtained for further statistical analysis and associated risk factors to seropositive flocks. For comparison, abortion rates between the vaccinated and non-vaccinated herds were compared. The overall seroprevalence of the unvaccinated herds was 33%, which can be considered as high when compared to other European regions. Associated risk factors such as herd size, management system, diet, and manure removal frequency were found statistically significant. However, no significant differences were found in the abortion rates between vaccinated and non-vaccinated flocks, indicating that other microorganisms could also cause abortions in goats on the region. Despite this, the seroprevalence of C. abortus is relatively high in this limited survey of goat herds and may pose a threat to both human and animals on the Canary Islands.

Doxycycline treatment for Dirofilaria immitis in dogs: impact on Staphylococcus aureus and Enterococcus antimicrobial resistance.

Doxycycline is an antibiotic that, in addition to the classic antibacterial use, is also prescribed to fight parasitic diseases, like heartworm disease in dogs. Despite the concern that the overuse of this antibiotic may decrease susceptibility of clinically important bacteria, the consequences of the prolonged doxycycline therapy in heartworm-infected dogs have never been studied before. We have analyzed the impact of this therapy on Staphylococcus aureus and Enterococcus antimicrobial resistance. In this study, 17 heartworm-infected dogs (10 that had completed the doxycycline treatment and 7 dogs that had not yet begun) were included. Twenty-four isolates of Staphylococcus aureus were obtained from two locations of each dog. After treatment, 73.3% of isolates were resistant to at least one antibiotic but only 22.2% of isolates before treatment. Most of doxycycline resistant isolates were obtained from dogs that have received treatment. Erythromycin resistance or intermediate susceptibility was detected in 45.6% of isolates, most of them from dogs after treatment. For Enterococci, 48 isolates were obtained from fecal samples (25 before treatment and 23 after treatment). Before treatment, 32% of isolates were resistant at least to one antibiotic while after, this data increase up to 65%. Comparing isolates before and after treatment, a clear increase in resistance to doxycycline (12% against 21.74%) and erythromycin (20% against 39.13%) was observed. Although the present work is a preliminary research, the results encourages the development of further studies to determinate the effect of prolonged doxycycline therapy on antimicrobial resistance.

Identification, antimicrobial susceptibility, and virulence factors of Enterococcus spp. strains isolated from Camels in Canary Islands, Spain.

This study investigated the presence of Enterococcus spp. strains in camel faeces, their virulence factors, and resistance to the antibiotics commonly used as therapy of enterococcal infections. One hundred and seventy three Enterococcus strains were isolated and identified to species level using polymerase chain reaction (PCR). Susceptibility to 11 antimicrobials was determined by disk diffusion method. Minimal Inhibitory Concentrations (MIC) of penicillin, ampicillin, vancomycin, teicoplanin, gentamicin, and streptomycin were all determined. Genes encoding resistance to vancomycin, tetracycline, and erythromycin as well as genes encoding some virulence factors were identified by PCR. Enterococcus hirae (54.3%) and Enterococcus faecium (25.4%) were the species most frequently isolated. None of the strains were resistant to vancomycin, teicoplanin, ampicillin or showed high level aminoglycoside resistance (HLAR). Strains resistant to rifampicin (42.42%) were those most commonly found followed those resistant to trimethoprim - sulfamethoxazole (33.33%). The genes tetM, tetL, vanC1, and vanC2-C3 were detected in some strains. Virulence genes were not detected. Monitoring the presence of resistant strains of faecal enterococci in animal used with recreational purposes is important to prevent transmission of those strains to humans and to detect resistance or virulence genes that could be transferred to other clinically important bacteria.

Stomoxys calcitrans as possible vector of Trypanosoma evansi among camels in an affected area of the Canary Islands, Spain.

INTRODUCTION: Trypanosoma evansi was first identified in the Canary Islands in 1997, and is still present in a small area of the Archipelago. To date, the disease has exclusively affected camel herds, and has not been detected in any other animal hosts. However potential vectors of Trypanosoma evansi must be identified. METHODS: One Nzi trap was placed on a camel farm located in the infected area for a period of one year. RESULTS: Two thousand five hundred and five insects were trapped, of which Stomoxys calcitrans was the sole hematophagous vector captured. CONCLUSIONS: Stomoxys calcitrans could be exclusively responsible for the transmission of Trypanosoma evansi among camels in the surveyed area, as other species do not seem to be infected by S. calcitrans in the presence of camels.