Antibiotic Susceptibility, Resistance Gene Determinants and Corresponding Genomic Regions in Lactobacillus amylovorus Isolates Derived from Wild Boars and Domestic Pigs.

Restrictions on the use of antibiotics in pigs lead to the continuous search for new probiotics serving as an alternative to antibiotics. One of the key parameters for probiotic bacteria selection is the absence of horizontally transmissible resistance genes. The aim of our study was to determine antibiotic susceptibility profiles in 28 Lactobacillus amylovorus isolates derived from the digestive tract of wild boars and farm pigs by means of the broth microdilution method and whole genome sequencing (WGS). We revealed genetic resistance determinants and examined sequences flanking resistance genes in these strains. Our findings indicate that L. amylovorus strains from domestic pigs are predominantly resistant to tetracycline, erythromycin and ampicillin. WGS analysis of horizontally transmissible genes revealed only three genetic determinants (tetW, ermB and aadE) of which all tetW and ermB genes were present only in strains derived from domestic pigs. Sequence analysis of coding sequences (CDS) in the neighborhood of the tetW gene revealed the presence of site-specific recombinase (xerC/D), site-specific DNA recombinase (spoIVCA) or DNA-binding transcriptional regulator (xre), usually directly downstream of the tetW gene. In the case of ermB, CDS for omega transcriptional repressor or mobilization protein were detected upstream of the ermB gene.

Microbial contamination of harvested colostrum on Czech dairy farms.

The objective of this study was to perform a quantitative and qualitative evaluation of microbial contamination of harvested colostrum on 39 Czech dairy farms. The study identified the proportion of colostrum samples that met the recommended goals for total plate count (TPC), total coliform count (TCC), and gram-negative noncoliform count (NCC), and evaluated the effect of the farm, breed, parity, season of the year, time of calving, and colostrum volume on these 3 microbiological parameters. Colostrum samples from cows (n = 1,241; 57.6% from Czech Fleckvieh, and 42.4% from Holstein breed) were collected on dairy farms between autumn of 2015 and spring of 2017. The samples were collected after the first milking directly from milking buckets. In 155 out of 1,241 colostrum samples (26 farms, 6 samples each, except 1 farm), the species of microorganisms obtained by culture were determined, and the findings were classified into 4 groups according to the probable source of contamination as follows: (1) normal inhabitants of bovine skin and mucosa, (2) fecal contaminants, (3) environmental contaminants, and (4) potential gram-positive mammary pathogens. Our results showed heavy microbial contamination of collected colostrum samples (TPC median = 408,000 cfu/mL; TCC median = 200 cfu/mL; NCC median = 80 cfu/mL). Only 28.4% of samples met the requirement for TPC (<100,000 cfu/mL), 88.2% for TCC (<10,000 cfu/mL), and 86.0% for NCC (<5,000 cfu/mL). Among the tested factors, we found that farm had a significant effect on all 3 microbiological parameters, volume of colostrum had an effect on TPC (the highest TPC in <3.0 L of colostrum), and season had an effect on TCC and NCC (higher TCC in summer than in autumn and winter; the highest NCC in summer and higher in autumn than in spring and winter). Our results showed that most microbes isolated from colostrum belonged to normal inhabitants of bovine skin and mucosa, fecal, or environmental contaminants (i.e., 82.6%, 81.9%, and 75.5% of colostrum samples, respectively). Potential gram-positive mammary pathogens were found in 13.5% of samples. Escherichia coli was isolated from 9.0% of colostrum samples, and Streptococcus uberis and Streptococcus parauberis were each isolated from 5.2% of samples. Our study showed high microbial contamination of colostrum collected on dairy farms. Therefore, better hygiene and sanitation around colostrum harvest should be addressed by farmers.

New sequence type ST3756 of Staphylococcus aureus subspecies anaerobius as the causative agent of abscessing lymphadenitis in sheep.

Morel's disease is a form of abscessing lymphadenitis of sheep and goats caused by Staphylococcus aureus subspecies anaerobius. In Europe and Africa, the disease is linked to S. aureus of multilocus sequence type 1464. In an outbreak recorded in 2015 in a flock of 530 animals in the district of Nymburk, Czech Republic, Europe, the causative agent was cultured and subsequently confirmed by Maldi-TOF. Neither antibiotic therapy nor surgical interventions met any success, although the strain isolated was found to be sensitive to antibiotics used. Vaccination and revaccination with inactivated autogenous vaccine administered subcutaneously was relatively successful. Subsequent multilocus sequence typing revealed the presence of new S. aureus sequence type 3756, different from 1464 in three out of seven genes typed. The isolate thus represents a new sequence type of Staphylococcus aureus ssp. anaerobius which should be considered as a causative agent of Morel's disease.