Characterization of Gallibacterium anatis Isolated from Pathological Processes in Domestic Mammals and Birds in the Czech Republic.

Gallibacterium anatis, recognized as a resident and opportunistic pathogen primarily in poultry, underwent investigation in unwell domestic mammals and birds. The study encompassed the mapping and comparison of G. anatis isolates, evaluation of their genetic diversity, and determination of their susceptibility to antimicrobials. A total of 11,908 clinical samples were analyzed using cultivation methods and MALDI-TOF. Whole-genome sequencing was performed on seven calf isolates and six hen isolates. Among mammals, G. anatis was exclusively detected in 22 young dairy calves, while among domestic birds, it was found in 35 individuals belonging to four species. Pathological observations in calves were predominantly localized in the digestive tract, whereas in birds, multi-organ infections and respiratory system infections were most prevalent. Distinct groups of genes were identified solely in calf isolates, and conversely, those unique to hen isolates were also recognized. Novel alleles in the multilocus sequence typing scheme genes and previously unidentified sequence types were observed in both calf and hen isolates. Antimicrobial susceptibility exhibited variation between bird and calf isolates. Notably, G. anatis isolates from calves exhibited disparities in genotype and phenotype compared to those from hens. Despite these distinctions, G. anatis isolates demonstrated the capability to induce septicemia in both species.

Sero-epidemiology of tick-borne encephalitis in small ruminants in the Czech Republic.

Tick-borne encephalitis virus (TBEV) is an arbovirus that causes severe infections in humans, and is endemic to large areas of Europe and Asia. Humans most commonly become infected with TBEV after a tick bite; however, alimentary infection can occur after consumption of unpasteurized dairy products. Milk from sheep and goats can be a source of alimentary TBE infections. In addition, sheep and goats are considered suitable sentinels for surveillance of TBEV-associated risks in endemic areas. Here we conducted a serological survey to determine the prevalence of TBEV infection among sheep and goats in the Czech Republic. In 2019-2020, a total of 310 serum samples were collected from sheep and 418 from goats, in 11 of the 14 administrative districts of the country. Sera were tested for the presence of TBEV-specific IgG by ELISA, and suspected results were validated using a virus neutralization test. Positive samples were identified in 56.7% of goat farms, and 82.4% of sheep farms, and in 9 of the 11 administrative districts examined. The seroprevalence was significantly higher among sheep (32.5%) than goats (19.7%) (p < 0.001). The present results indicate that sheep and goats have a relatively high rate of exposure to TBEV-infected ticks in most of the administrative districts of the Czech Republic. These findings confirm the usefulness of serological testing in small ruminants to determine and monitor the risk of TBEV infection in humans.

Gordonia species as a rare pathogen isolated from milk of dairy cows with mastitis.

While Gordonia species have long been known to cause severe inflammation in humans, the pathogenic effects of Gordonia species in veterinary medicine have rarely been described. Between 2010 and 2019, we collected microorganisms of the genus Gordonia isolated from milk samples from dairy cows with mastitis. We describe the growth properties of these microorganisms and their prevalence, virulence factors and susceptibility to antimicrobial agents. From 31,534 quarter milk samples processed by standard culture methods, 27 isolates of Gordonia species (0.086% prevalence) were identified by a molecular phenotyping method. The isolates originated from 17 farms in 12 districts of the Czech Republic. Twenty-one isolates were tested for susceptibility to 7 antimicrobials by the disc diffusion method. Notably, 100% of these isolates were susceptible to streptomycin and neomycin, 85.7% to cefovecin and tetracycline, 76.2% to penicillin G, 47.6% to trimethoprim/sulfamethoxazole and 0% to clindamycin. The species was determined to be Gordonia paraffinivorans by whole genome sequencing for 9 isolates (from 8 farms in 7 districts). These isolates showed the highest similarity to two reference strains from the environment. In all these isolates, we identified genes encoding virulence factors that are very similar to genes encoding virulence factors expressed in Mycobacterium tuberculosis and Mycobacterium smegmatis. However, genome analysis revealed 61 unique genes in all 9 sequenced isolates.

Streptococcus suis Isolates-Serotypes and Susceptibility to Antimicrobials in Terms of Their Use on Selected Repopulated Czech Pig Farms.

Streptococcus suis represents a primary health problem (such as meningitis, septicemia and arthritis in piglets and fatteners) in the swine industry worldwide and also an emerging zoonotic pathogen. In the Czech Republic, many pig farms repopulated their herds over the past decades to reduce morbidity and minimize treatment. The study analysed serotypes, sequence types and antimicrobial susceptibility in 39 S. suis isolates obtained from organs of diseased pigs from selected 16 repopulated farms with a history of S. suis-associated diseases and routine antimicrobial treatment with tulathromycin and/or amoxicillin. The analysis revealed diversity of collected isolates with regular occurrence of more than three serotypes per farm. The serotypes identified were 1/2 and 7, each in six isolates, followed by serotype 2 and 3 found in five isolates each, other serotypes were less frequent. Seven isolates were not typable by multiplex PCR and we also found sequence type of unknown type in thirteen isolates. The majority of S. suis isolates were resistant to clindamycin (n = 31), tetracycline (n = 29) and tilmicosin and tulathromycin (n = 28). On the other hand, with the exception of two isolates that were intermediately susceptible to penicillin and ampicillin, all isolates were susceptible to all three tested subgroups of beta-lactam antibiotics.

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.