Bacteria culturing and isolation under field conditions of Moukalaba-Doudou National Park, Gabon, and preliminary survey on bacteria carrying antibiotic resistance genes

Abstract

Culturing the intestinal bacteria of wild animals is a difficult task under field conditions, although isolation and characterization of bacteria are absolutely necessary to evaluate the transmission of bacteria from human to animal or vice versa. We have developed a protocol for intestinal bacteria culturing from feces of wild animals such as gorillas in Moukalaba-Doudou National Park in Gabon, where no laboratory settings were available. The prevalence of resistance genes in Enterobacteriaceae isolates estimated by real time PCR array was higher for aminoglycosides resistance, followed by tetracycline resistance, except for possible naturally occurring β-lactam resistance. The detection level of resistance genes was higher for isolates from humans than those from gorillas. Occasional monitoring of this prevalence may help to measure the intensity of introduction of humanborne bacteria to wildlife.

Characterization of ESBL-Producing Enterobacteria from Fruit Bats in an Unprotected Area of Makokou, Gabon

“…A cotton swab was rotated inside the bat rectum and was immediately discharged into 5 mL of sterile water. One millilitre of liquid sample was inoculated into BacT/Alert blood culture media (bioMérieux, Auvergne-Rhône-Alpes, France) according to a previously established protocol [30]. After incubation, enterobacteria present in the medium produced CO 2 during their growth phase.…”

“…ESBL types temoneira (TEM), sulfhydryl variable (SHV), and cefotaximase-Munich (CTX-M) are the most common ESBLs found in humans, livestock, and wildlife [6]. The global spread of ESBL-producing Enterobacteriaceae, particularly CTX-M-15, creates serious therapeutic challenges [10,[21][22][23][24][25][26][27][28][29][30][31][32].…”

“…In Gabon, CTX-M-15 has only been identified in ESBL-producing Enterobacteriaceae isolates from patients in the Albert Schweitzer Hospital in Lambaréné [31] and in poultry [32], but not in wildlife. Although previous studies on antibiotic resistance in Gabon have already described some phenotypes and resistance genes in wild terrestrial mammals [21,29,30], nothing is known about their dissemination in the environment. We hypothesized that bats could participate in the dissemination of antibiotic resistance in wildlife [14].…”

Isolation of multiple drug-resistant enteric bacteria from feces of wild Western Lowland Gorilla (<i>Gorilla gorilla gorilla</i>) in Gabon

“…In fact, there were several surveys regarding the antibiotic resistance of infectious bacteria in 1997 and 2009 in a hospital located adjacent to MDNP that clearly indicated a significant progress in the resistance of diarrheagenic E. coli isolates to second-generation cephalosporin, third-generation cephalosporin, carbapenem, aminoglycosides and the new quinolones [ 27 , 29 ]. The multidrug-resistant Achromobacter presently found in gorillas together with previously suggested resistant Enterobacter and Klebsiella [ 22 ] in wild western lowland gorillas may suggest that gorillas in Gabon have been exposed to human-borne resistant bacteria. Although the prevalence of multi-drug resistant bacteria was not high ( Table 1 ), wild animals like gorilla in MDNP may have received the stronger anthropological influences comparing to those in Lopé National Park in terms of transmission of bacteria.…”

“…In a preliminary survey, we have detected a range of antibiotic-resistant genes from the fecal Enterobacteriaceae of wild gorillas in MDNP [ 22 ]. The most prevalent genes were responsible for resistance against β-lactam antibiotics.…”