The prevalence of methicillin resistance in staphylococci has been increasing globally and is currently one of the major public health concerns. In particular, treating infections caused by staphylococci with acquired antimicrobial resistance is problematic, as their treatment is more difficult. The resistance is found both in human and animal staphylococcal strains. Methicillin-resistant staphylococci (MRS) have also been increasingly reported in wildlife. In Arab countries, MRS has been detected in food producing animals and food products; however, the risk this poses is somewhat unclear, and still a significant lack of information on the trend and distribution of these pathogens in these countries, which have a specific ecosystem (desert) and traditions (Muslim culture). In this manuscript, we aim to provide an overview of the prevalence and the major MRS clonal lineages circulating in these specific countries and compare to them other situations with different ecosystems and cultures.

Dromedary camels are an important source of food and income in many countries. However, it has been largely overlooked that they can also transmit antibiotic-resistant bacteria. The aim of this study was to identify the Staphylococcaceae bacteria composition of the nasal flora in dromedary camels and evaluate the presence of methicillin-resistant Mammaliicoccus (MRM) and methicillin-resistant Staphylococcus (MRS) in dromedary camels in Algeria. Nasal swabs were collected from 46 camels from seven farms located in two different regions of Algeria (M’sila and Ouargla). We used non-selective media to determine the nasal flora, and antibiotic-supplemented media to isolate MRS and MRM. The staphylococcal isolates were identified using an Autoflex Biotyper Mass Spectrometer (MALDI-TOF MS). The mecA and mecC genes were detected by PCR. Methicillin-resistant strains were further analysed by long-read whole genome sequencing (WGS). Thirteen known Staphylococcus and Mammaliicoccus species were identified in the nasal flora, of which half (49.2%) were coagulase-positive staphylococci. The results showed that four out of seven farms were positive for MRS and/or MRM, with a total of 16 isolates from 13 dromedary camels. The predominant species were M. lentus, S. epidermidis, and S. aureus. Three methicillin-resistant S. aureus (MRSA) were found to be ST6 and spa type t304. Among methicillin-resistant S. epidermidis (MRSE), ST61 was the predominant ST identified. Phylogenetic analysis showed clonal relatedness among M. lentus strains, while S. epidermidis strains were not closely related. Resistance genes were detected, including mecA, mecC, ermB, tet(K), and blaZ. An SCCmec type VIII element was found in a methicillin-resistant S. hominis (MRSH) belonging to the ST1 strain. An SCCmec-mecC hybrid element was detected in M. lentus, similar to what was previously detected in M. sciuri. This study highlights that dromedary camels may be a reservoir for MRS and MRM, and that they contain a specific set of SCCmec elements. This emphasizes the need for further research in this ecological niche from a One Health perspective.

Sarcocystis spp. (Apicomplexa: Coccidia) are obligate heteroxenous protozoa that infect a wide range of host species. Transmission follows a predator-prey cycle involving an intermediate host (IH) and a definitive host (DH). For many species, only IHs have been identified, while DHs remain unknown. DHs can be infected with multiple Sarcocystis spp. at the same time, which complicates species identification. We aimed to determine the prevalence and species diversity of Sarcocystis infections in free-ranging wolves in Switzerland using both coprological and molecular methods. A further goal was to evaluate the utility of Third-generation sequencing for resolving mixed infections. A total of 87 wolf intestinal content samples were collected between 2017 and 2023 and analyzed coproscopically by a sedimentation-flotation method. Sarcocystis oocysts/sporocysts were detected in 76 % (66/87). DNA was obtained from 57/66 positive samples and 55/57 resulted positive in a Sarcocystis 18S rRNA screening PCR. Additionally, mitochondrial cytochrome c oxidase subunit I (COI) gene PCR and a real-time PCR targeting S. cruzi were performed. PCR products from conventional PCRs were submitted for Sanger sequencing. Monoinfections were identified in 16 % (9/55) and mixed infections in 84 % (46/55) of the samples. A subset of five samples was analyzed by Third-generation sequencing (Pacific Biosciences) of the 18S rRNA full-length and COI fragment PCR products. BLAST and phylogenetic analysis were used to validate taxonomic classification. Molecular analysis identified nine known Sarcocystis species: S. tenella, S. arieticanis, S. capreolicanis, S. linearis, S. gracilis, S. cruzi, S. capracanis, S. iberica, and S. venatoria. Newly developed pipelines for the Third-generation sequencing data provided high-resolution species-level identification in samples with mixed infections. These findings confirm the Eurasian wolf as natural DH for multiple Sarcocystis species for the first time, including S. linearis, S. iberica, and S. venatoria. Further complementary studies on prey species are needed to clarify host-parasite dynamics.

Nine Gram-positive, non-motile, facultatively anaerobic cocci, designated EM39Eᵀ, JEK85, 18KM676, 21M1142, 18KM445^T, 18KM444, 18KM245, 18KM583 and 21KM1573, were isolated from diverse animal material, including horse and pig skin, bovine mastitis milk and feline urine from a urinary tract infection. Phylogenomic analysis based on amino-acid alignment obtained from translated whole-genome sequences; digital DNA–DNA hybridization (dDDH); 16S rRNA, dnaJ, hsp60, rpoB and sodA gene comparison; and MALDI-TOF MS spectral profiles placed the strains within the Macrococcus caseolyticus clade. They were closely related to Macrococcus bohemicus CCM 7100ᵀ, Macrococcus epidermidis CCM 7099ᵀ, Macrococcus goetzii CCM 4927ᵀ and Macrococcus capreoli DSM 113939^T. The novel species exhibited 22.30% dDDH and 79.49% average nucleotide identity (ANI) to each other, <24% dDDH and <82% ANI to their closest relatives, values below established species delineation thresholds, confirming their classification as two distinct novel species. Chemotaxonomic analysis identified, in both species, diphosphatidylglycerol, phosphatidylglycerol, aminolipid, glycolipids and phosphoglycolipids, as the major polar lipids. Fatty acid profiling revealed C_14 : 0, C_16 : 1 ω11c, C_16 : 0 and C_18 : 1 ω13c as predominant components, with antesio-C_15 : 0 in one species. The peptidoglycan type was A3α l-Lys–Gly2. The novel species can be distinguished from their closest relatives by their ability to grow in 12.5% NaCl. EM39Eᵀ, JEK85, 18KM676 and 21M1142 can be distinguished by positive reaction to d-ribose, and 18KM445^T, 18KM444, 18KM245, 18KM583 and 21KM1573 by the absence of α-glucosidase activity and acid production from methyl-β-d-glucopyranoside and maltose. The two novel species can be differentiated from each other by d-ribose metabolism and methyl-β-d-glucopyranoside hydrolysis. 18KM445^T, 18KM444, 18KM245, 18KM583 and 21KM1573 formed a distinct cluster of strains, designated Macrococcus equi sp. nov., with strain 18KM445ᵀ (=DSM 118744ᵀ; =CCOS 2124ᵀ; =CCM 9438ᵀ) as the proposed type strain. The second cluster, containing strains EM39Eᵀ, JEK85, 18KM676 and 21M1142, was designated Macrococcus animalis sp. nov., with strain EM39Eᵀ (=DSM 118743ᵀ; =CCOS 2125ᵀ; =CCM 9439ᵀ) as the proposed type strain.

Methicillin-resistant Staphylococcus aureus (MRSA) sequence type 630 (ST630) is an emerging clone causing infections in humans. We report the complete genomes of three MRSA ST630 from cattle. The 2.8 Mbp genomes contained several antimicrobial resistance and virulence genes and were related to those of previously detected human-associated MRSA ST630.

Camel milk is a valuable food choice, representing a primary need in the diet of people in dry zones. Camel milk is rich in nutrients, which makes it a favorable environment for the development of microorganisms. Staphylococcus and Listeria are significant opportunistic pathogens in humans, dairy cattle, and camels. The presence of these bacteria could present a potential public health issue. In the present study, 20 milk samples collected from camel farms in M’sila were investigated for the presence of Staphylococcus spp. and Listeria spp. Staphylococcus enumeration, and a search for Listeria spp. was performed according to the recommendations of ISO 6888-1(2004) and ISO 11290-1(2017) methods, respectively. The results show a contamination prevalence of 62% of staphylococcus spp. with an estimated average bacterial load of 2.7.102 cfu/mL, while for Listeria spp., only three samples were positive, with a prevalence of 14.28%. For each species, identification using API Listeria strips confirmed the presence of Listeria grayi, Listeria innocua, and Listeria seeligeri species, but no Listeria monocytogenes were recovered in these samples. According to the current results, we could conclude that the percentage of contamination with Staphylococcus in the tested camel milk samples was relatively high compared to the level of contamination with Listeria. There is no Algerian regulation setting microbiological criteria for raw camel milk; however, these results suggest that the hazard to the consumer cannot be excluded.