Resistant Escherichia coli isolated from wild mammals from two rescue and rehabilitation centers in Costa Rica: characterization and public health relevance.

This study aimed to characterize the antimicrobial resistance (AMR) and virulence profiles of 67 Escherichia coli isolates obtained from faecal samples of 77 wild mammals from 19 different species, admitted in two rescue and rehabilitation centers in Costa Rica. It was possible to classify 48% (n = 32) of the isolates as multidrug-resistant, and while the highest resistance levels were found towards commonly prescribed antimicrobials, resistance to fluoroquinolones and third generation cephalosporins were also observed. Isolates obtained from samples of rehabilitated animals or animals treated with antibiotics were found to have significantly higher AMR levels, with the former also having a significant association with a multidrug-resistance profile. Additionally, the isolates displayed the capacity to produce α-haemolysins (n = 64, 96%), biofilms (n = 51, 76%) and protease (n = 21, 31%). Our results showed that AMR might be a widespread phenomenon within Costa Rican wildlife and that both free-ranging and rehabilitated wild mammals are potential carriers of bacteria with important resistance and virulence profiles. These results highlight the need to study potential sources of resistance determinants to wildlife, and to determine if wild animals can disseminate resistant bacteria in the environment, potentially posing a significant threat to public health and hindering the implementation of a "One Health" approach.

REPRODUCTIVE CYCLE STAGE ASSESSMENT IN THE TWO-TOED SLOTH (CHOLOEPUS HOFFMANNI), PRELIMINARY RESULTS.

The number of two-toed sloths (Choloepus hoffmanni) has significantly decreased in the last years. Deepening the knowledge of this tropical mammal's reproductive physiology is essential to improve captive breeding within conservation programs for this species. However, several aspects of its reproductive biology remain unexplored and have not been described sufficiently. The aim of this work was to describe the estrous cycle and reproductive physiology of two adult female C. hoffmanni by vaginal cytology, appearance of the external genitalia, and behavior. Vaginal cytology assay showed that the average duration of the estrous cycle was 15.1 ± 4.53 d. Positive correlations (P < 0.05) were found between the peak presence of superficial cells (estrous phase) and four parameters: aggressive behavior, pursuing behavior, vulvar swelling, and vaginal discharge. This pilot study, conducted on just two animals, forms a basis for a study design that may be employed for a more comprehensive assessment of the two-toed sloth reproductive physiology and behavior.

The Speaker Method: A Novel Release Method for Offspring Mammals and 5-Year Study on Three Costa Rican Mammals.

Nowadays, wild animals are threatened by humans, with the number of species and individuals decreasing during recent years. Wildlife rescue centers play a vital role in the conservation of wildlife populations. This study aims to describe a new release technique, the Speaker Method, to rescue and facilitate the reunion of different baby mammals that arrived at a wildlife rescue center with their mothers within their natural habitat, avoiding the need for captivity. This method is based on a recorded baby's cry played on a speaker to make a "call effect" in the mother. The efficacy of the Speaker Method for babies' reunion with their mothers was 45.8% in Hoffmann's two-toed sloths (Choloepus hoffmanni) and 91.9% in brown-throated sloths (Bradypus variegatus). Among the mantled howler monkeys (Alouatta palliata), 50% of the babies could be released using this new technique. The findings suggest that the method could be helpful in the early release of young individuals, highlighting higher release outcomes in these three species compared to traditional nursery care provided by human caretakers, who face inherent difficulties in raising young animals without their mothers.

Outbreaks of Acute Necrotic Enteritis and Haemolytic Crisis and Renal Disease Associated with Clostridium perfringens in Captive Toucans (Ramphastos spp).

Four toco toucans (Ramphastos toco), one channel-billed toucan (Ramphastos vitellinus) and one white-throated toucan (Ramphastos tucanus) died in two disease outbreaks in the same aviary in 2011 and 2016. Post-mortem examination revealed diffuse necrotic enteritis (NE) as the cause of death of five of these six birds. Clostridium perfringens was identified by culture and real-time multiplex PCR for C. perfringens α-, ß-, ε- and ι-toxin genes in ligated intestine of one toucan from each outbreak. At another aviary, two keel-billed toucans (Ramphastos sulfuratus) died peracutely from severe haemolytic crisis with haemoglobinaemic nephrosis and cholestasis and acute tubulointerstitial nephritis. Mild NE was present in these birds and C. perfringens was demonstrated in liver by bacterial culture and real-time multiplex PCR for C. perfringens α-, ß-, ε- and ι-toxin genes. To the authors' knowledge, this is the first description of outbreaks of NE associated with C. perfringens in captive toucans. Although haemolytic crisis has been reported in humans with C. perfringens type A septicaemia and hepatic abscesses, this presentation appears not to have been described in C. perfringens infections in toucans or other avian species. The factors causing C. perfringens proliferation and disease in the toucans were not identified. PCR for C. perfringens NetB toxin and enterotoxin genes performed retrospectively on one of the C. perfringens isolates from the second outbreak and on paraffin-embedded tissues from one dead toucan from the first outbreak was negative. With the current C. perfringens toxin typing scheme, C. perfringens type A was identified in the first two outbreaks.

TRAUMATIC (FOREIGN BODY) PERICARDITIS IN A TOCO TOUCAN (RAMPHASTOS TOCO).

An approximately 10-yr-old, captive-born female toco toucan ( Ramphastos toco ) was presented due to an acute onset of depression and apathy. On visual and physical examination, it showed an abnormal posture and dehydration, respectively. Serum biochemistry revealed hyperuricemia (39.4 mg/dl) and elevated glutamic oxaloacetic transaminase (GOT; 1,050 U/L). Radiographs demonstrated an enlargement of the cardiac silhouette. The bird died 7 days after presentation, despite treatment with enrofloxacin, allopurinol, a preparation of hepatorenal protectors, and complex B vitamins with dextrose. Necropsy revealed severe fibrinohemorrhagic pericarditis with a 15 mm long and 2.5 mm diameter, rigid foreign body in the pericardial exudate. Microscopically, this foreign body was of vegetal origin.

Bilateral Renal Tubular Neoplasm in a Channel-billed Toucan (Ramphastos vitellinus).

An adult male channel-billed toucan (Ramphastos vitellinus) was presented with a history of weakness, dyspnea, and severe dilatation of the coelomic cavity, which was caused by accumulation of serohemorrhagic fluid. Radiographs revealed increased radiodensity and thickness of the descending aorta and a pectoral mass, and blood test results revealed anemia, hypocalcemia, hypoproteinemia, and hyperuricemia. On ultrasound examination, a hyperechoic enlarged soft tissue mass was found in the caudodorsal region of the coelom. The bird did not respond to supportive care and died. Postmortem examination revealed severe, bilateral nephromegaly due to multifocal to coalescing renal tubular adenomas (adenomatosis), which was complicated with renal gout and soft tissue mineralization. Relevant concurrent diseases included hepatic hemochromatosis, subcutaneous cestodiasis with cellulitis, and systemic amyloidosis. There are few documented cases of neoplasms in ramphastid birds and to our knowledge, this is the first report of a renal neoplasm in a channel-billed toucan.