-
Pathogens (Basel, Switzerland) Feb 2023Antimicrobials are added to semen extenders to inhibit the growth of bacteria that are transferred to the semen during collection. However, this non-therapeutic use of...
Antimicrobials are added to semen extenders to inhibit the growth of bacteria that are transferred to the semen during collection. However, this non-therapeutic use of antimicrobials could contribute to the development of antimicrobial resistance. The objective of this study was to determine changes in the antibiotic susceptibility of vaginal microbiota after artificial insemination. Swabs were taken from the vagina of 26 mares immediately before artificial insemination and again 3 days later. Bacteria isolated from the vagina at both time points were subjected to antibiotic susceptibility testing and whole-genome sequencing. In total, 32 bacterial species were identified. There were increases in the resistance of to trimethoprim ( = 0.0006), chloramphenicol and ( = 0.012) tetracycline ( = 0.03) between day 0 and day 3. However, there was no significant effect of exposure to antibiotics in semen extenders with respect to the resistance of and ( > 0.05). Whole-genome sequencing indicated that most phenotypic resistance was associated with genes for resistance. These results indicate that the resistance patterns of vaginal bacteria may be affected by exposure to antibiotics; therefore, it would be prudent to minimize, or preferably, avoid using antibiotics in semen extenders.
PubMed: 36986297
DOI: 10.3390/pathogens12030375 -
Journal of Dairy Science Feb 2018Subclinical mastitis causes an increase in milk somatic cell count (SCC) and can lead to reduced milk production and early culling. In many countries, non-aureus...
Subclinical mastitis causes an increase in milk somatic cell count (SCC) and can lead to reduced milk production and early culling. In many countries, non-aureus staphylococci (NAS) is the most common bacterial finding in subclinical mastitis of dairy cows. New methodology makes it possible to identify NAS species, but knowledge about the epidemiology is limited. The objective of this project was to improve advisory services for mastitis control by investigating associations between NAS and SCC, milk production, and persistence of intramammary infections (IMI). Farmers who had sent milk samples to the Swedish National Veterinary Institute (Uppsala, Sweden) were asked to participate if NAS was identified in the samples. Participating farmers were asked to resample all udder quarters of the cow once within 1 mo. Regression models were used to investigate associations between NAS and cow factors, udder quarter California mastitis test and SCC, and persistence of IMI. Associations with cow composite milk yield and SCC were also investigated. In total, 671 cows from 201 herds were enrolled in the study, and 19 NAS species were identified, of which the 4 most common were Staphylococcus epidermidis, Staphylococcus simulans, Staphylococcus chromogenes, and Staphylococcus haemolyticus. Persistent IMI was more common in udder quarters with Staphylococcus hyicus and S. simulans and less common in those with Staphylococcus saprophyticus IMI. β-Lactamase production by the different NAS species varied from 0 to 100%. There was a significant association between NAS species and California mastitis test and SCC of udder quarters, and this varied depending on parity. The cow composite milk SCC at the test milking before the initial sample was taken differed significantly with NAS species, but not at the subsequent test milking. Milk yield-at the test milking before or after the initial sample-did not differ significantly for NAS species. There were no significant associations between milk yield or SCC and persistent NAS IMI. In conclusion, the NAS species affects SCC and persistent IMI differently but not milk yield.
Topics: Animals; California; Cattle; Cell Count; Dairying; Female; Mastitis, Bovine; Milk; Parity; Pregnancy; Staphylococcus; Staphylococcus epidermidis; Staphylococcus haemolyticus; Staphylococcus hyicus; Sweden; beta-Lactamases
PubMed: 29174143
DOI: 10.3168/jds.2017-13467 -
Central European Journal of Public... Jun 2022This work aimed to determine the representation and resistance of bacteria belonging to the genus Staphylococcus and Enterococcus on inanimate surfaces of two selected...
OBJECTIVES
This work aimed to determine the representation and resistance of bacteria belonging to the genus Staphylococcus and Enterococcus on inanimate surfaces of two selected workplaces of the University Hospital of L. Pasteur in Košice (UHLP) and to investigate their importance in the hospital environment. The men's ward of the Department of Internal Medicine (DIM) and the Department of Anaesthesiology and Intensive Care (DAIC) were chosen.
METHODS
Using sterile sampling kits, a total of 182 swabs were collected from the inanimate surfaces of both UHLP workplaces. The swabs were then transported to a microbiological laboratory and inoculated onto sterile culture media (blood agar containing 5% ram erythrocytes). After culturing (24-48 hours, in a thermostat at constant temperature 37 °C), bacterial colonies were identified by mass spectrometry on a MALDI TOF MS. Bacteria belonging to the genera Staphylococcus and Enterococcus were subsequently separated from the spectrum of identified bacteria. Nosocomial significant strains of staphylococci (Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus aureus) and all isolated enterococci were subjected to susceptibility testing for selected antibiotics using the disk diffusion method - E-tests.
RESULTS
Several members of the genus Staphylococcus were identified from the inanimate surfaces of both workplaces. These were mainly coagulase-negative strains - Staphylococcus epidermidis (45), Staphylococcus capitis (34), Staphylococcus haemolyticus (20), Staphylococcus hominis (45), Staphylococcus pasteuri (2), Staphylococcus sroph (1), Staphylococcus simulans (3), and Staphylococcus warneri (4). Staphylococcus aureus strains were also identified (2). Nosocomial significant isolates were tested for susceptibility to the antibiotics cefoxitin (FOX) and oxacillin (OXA). Two members of the genus Enterococcus - Enterococcus faecium (7) and Enterococcus faecalis (8) were isolated. All strains were subject to vancomycin susceptibility testing using the disk method.
Topics: Animals; Anti-Bacterial Agents; Bacteria; Cross Infection; Enterococcus; Hospitals; Humans; Male; Microbial Sensitivity Tests; Sheep; Staphylococcal Infections; Staphylococcus
PubMed: 35841227
DOI: 10.21101/cejph.a7241 -
Environmental Research Apr 2019The airborne fungal and bacterial species present in pig farm dust have not been well characterised even though these bioaerosols are known to cause inflammation and...
The airborne fungal and bacterial species present in pig farm dust have not been well characterised even though these bioaerosols are known to cause inflammation and other airway maladies. In this study, the microbial species and composition in airborne dust within and between pig farms were investigated. Passively sedimenting dust from six pig farms were collected using electrostatic dust collectors. The bacterial and fungal species were identified using matrix-assisted laser desorption-ionisation time-of-flight mass spectrometry (MALDI-TOF MS) and next generation sequencing (NGS). Dust samples taken within the same stable section revealed high resemblance and stability. Constrained statistical analysis of the microbial community compositions indicated that the types of stable did not appear to have a great effect on the bacterial and fungal β-diversity. In contrast to this, the farm from which samples were taken appeared to have the greatest effect on the bacterial β-diversity, but this trend was not observed for the fungal β-diversity. The most common bacteria and fungi according to NGS data were anaerobes typically associated with the pig intestinal tract and yeasts respectively. Bacterial sedimentation varied at a rate between 10 and 10 CFU/m/day, with the most common species after aerobic incubation being Aerococcus viridans and Staphylococcus equorum, while Clostridium perfringens and Staphylococcus simulans were the most common species after anaerobic incubation. A total of 28 different species of bacteria and fungi were classifiable as pathogens. In conclusion, the biodiversity in pig farm dust shows a high diversity of bacterial species. However, samples from the same stable section resembled each other, but also different sections within the same farm also resembled each other, thus indicating a high degree of community stability in the dust source. In regards to fungal identification, the biodiversity was observed to be similar between samples from different stable sections and farms, indicating a higher degree of similarities in the mycobiomes found across pig farms studied.
Topics: Air Microbiology; Animals; Bacteria; Biodiversity; Dust; Environmental Monitoring; Farms; Fungi; Swine
PubMed: 30771719
DOI: 10.1016/j.envres.2019.01.008 -
Journal of Dairy Science Jul 2018The most common pathogens causing intramammary infections (IMI) in dairy goats are staphylococci. Gene sequencing has been the reference method for identification of...
The most common pathogens causing intramammary infections (IMI) in dairy goats are staphylococci. Gene sequencing has been the reference method for identification of staphylococcal species, but MALDI-TOF mass spectrometry could represent a rapid and cost-effective alternative method. The objectives were to evaluate the typeability and accuracy of partial gene sequencing and MALDI-TOF for identifying staphylococci isolated from caprine milk samples, and to evaluate the relationship between staphylococcal species IMI, milk somatic cell score (SCS), and milk yield (MY). A composite (goat-level) milk sample was collected from all 940 lactating goats in a single herd. Dairy Herd Information Association test-day data for parity, days in milk, SCS, and MY were retrieved from Dairy Herd Information Association records. Milk samples were cultured on Columbia blood agar, and isolates from samples that yielded a single colony type of a presumptively identified Staphylococcus spp. were identified by PCR amplification and partial sequencing of rpoB, tuf, or 16S-rRNA, and MALDI-TOF. Mixed linear models were used to evaluate the relationship between staphylococcal IMI, SCS, and MY. The goat-level prevalence of staphylococcal IMI based on isolation of a single colony type was 24.4% (213/874). Seventeen goats had a contaminated sample. Among the remaining goats (n = 857), the most common species causing single colony-type IMI were Staphylococcus simulans (7.9%), Staphylococcus xylosus (3.5%), Staphylococcus caprae (3.6%), Staphylococcus chromogenes (2.9%), and Staphylococcus epidermidis (2.2%). The typeability of staphylococcal isolates with partial housekeeping gene sequence analysis (rpoB, complemented by tuf and 16S as needed) was 97.7%. The typeability and accuracy of MALDI-TOF were 84 and 100%, respectively. Overall, only Staphylococcus chromogenes IMI was associated with a higher SCS than goats with no growth. After adjusting for parity and stage of lactation, staphylococcal IMI status was not significantly associated with MY. For the staphylococci isolated from goats in this herd, MALDI-TOF proved an accurate method of speciation with a relatively high typeability. An association between staphylococcal IMI, SCS, and MY was not defined using goat-level data with the exception of S. chromogenes IMI, which was associated with a higher SCS than goats with no growth.
Topics: Animals; Cattle; Female; Goat Diseases; Goats; Lactation; Mastitis; Milk; Pregnancy; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Staphylococcal Infections; Staphylococcus
PubMed: 29705416
DOI: 10.3168/jds.2017-14224 -
Pathogens (Basel, Switzerland) Oct 2022The objectives of the work were (a) to compare the efficacy of two routes for antibiotic administration in the treatment of mastitis in ewes and (b) to assess the...
The objectives of the work were (a) to compare the efficacy of two routes for antibiotic administration in the treatment of mastitis in ewes and (b) to assess the potential importance of the timing of the initiation of the therapeutic regime on the outcome of the treatment. The ewes were allocated at random into three equal groups; intramammary inoculation with a isolate was performed, and clinical mastitis developed. The ewes in groups T1 ( = 6) and T2 ( = 6) were treated by the intramammary administration of ampicillin and dicloxacillin (two administrations with a 12-h interval). The ewes in group T3 ( = 6) were treated by the intramuscular injection of ampicillin and dicloxacillin (0.75 mL per 10 kg bodyweight, three injections with a 24-h interval). In the ewes in groups T1 and T3, treatment started immediately when the clinical signs of mastitis were first detected during the periodic examination of the ewes; in the ewes in group T2, treatment started 24 h after the clinical signs of mastitis were first detected. The animals were monitored clinically; mammary secretion samples were collected for bacteriological and cytological examinations. The median duration of the clinical signs was 4.75, 7.13, and 4.75 d for T1, T2, and T3; significant differences in clinical severity between the groups were seen until the 7th day post-treatment. The median duration of bacterial recovery was 3.25, 8.00, and 8.00 d for T1, T2, and T3; significant differences in the frequency of bacterial recovery between the groups were seen until (64.1%, 94.9%, and 96.2% of the samples) and after (2.9%, 16.7%, and 11.8%) the 7th day post-treatment. The median period required for the complete cure (clinical, bacteriological, and cytological) was shorter in the T1 than in the T2 and T3 ewe groups: 20.0, 32.0, and 24.5 d, respectively. The findings cover a gap in the available literature regarding the treatment of clinical mastitis in ewes. Early treatment resulted in the improved cure of the infection. The comparison of the intramammary and injectable routes for antibiotic administration indicated some benefit for the former, primarily in the post-treatment somatic cell counts.
PubMed: 36297221
DOI: 10.3390/pathogens11101164 -
Journal of Dairy Science Jan 2017Coagulase-negative staphylococci (CNS) have become the main pathogens causing bovine mastitis in recent years. A huge variation in species distribution among herds has...
Coagulase-negative staphylococci (CNS) have become the main pathogens causing bovine mastitis in recent years. A huge variation in species distribution among herds has been observed in several studies, emphasizing the need to identify subgroup- and species-specific herd-level factors to improve our understanding of the differences in ecological and epidemiological nature between species. The use of bulk milk samples enables the inclusion of a large(r) number of herds needed to identify herd-level risk factors and increases the likelihood of recovering enough isolates per species needed for conducting subgroup- and, eventually, species-specific analyses at the same time. This study aimed to describe the prevalence and distribution of CNS species in bulk milk samples and to identify associated subgroup- and species-specific herd-level factors. Ninety percent of all bulk milk samples yielded CNS. Staphylococcus equorum was the predominant species, followed by Staphylococcus haemolyticus and Staphylococcus epidermidis. A seasonal effect was observed for several CNS species. Bulk milk samples from herds with a loose-pack or a tiestall housing system were more likely to yield CNS species compared with herds with a freestall barn, except for S. epidermidis, Staphylococcus simulans, and Staphylococcus cohnii. In September, herds in which udders were clipped had lower odds of yielding Staphylococcus chromogenes, S. simulans, and Staphylococcus xylosus, the CNS species assumed to be most relevant for udder health, in their bulk milk than herds in which udder clipping was not practiced. Bulk milk of herds participating in a monthly veterinary udder health-monitoring program was more likely to yield these 3 CNS species. Herds always receiving their milk quality premium or predisinfecting teats before attachment of the milking cluster had lower odds of having S. equorum in their bulk milk. Herds not using a single dry cotton or paper towel for each cow during premilking udder preparation were more likely to have S. cohnii-positive bulk milk. Herds in which flushing with hot water or steam of the milking cluster after having milked a cow with a (sub)clinical mastitis was applied, were less likely to yield S. simulans, S. haemolyticus, and S. cohnii in their bulk milk. Always wearing gloves during milking decreased the odds of having Staphylococcus devriesei-positive bulk milk. Tap water from the public drinking system used as drinking water increased the odds of yielding S. simulans in the bulk milk. In conclusion, CNS are highly prevalent in bulk milk and might originate from the environment for some species (we hypothesize this is true for S. equorum or S. cohnii), or from within the udder (e.g., for S. simulans). Studies collecting bulk milk and quarter milk samples at the same time along with environmental samples are needed to determine the exact origin of the different (subgroups of) CNS species present in bulk milk using strain-typing techniques.
Topics: Animals; Cattle; Coagulase; Female; Mastitis, Bovine; Milk; Prevalence; Risk Factors; Staphylococcal Infections; Staphylococcus
PubMed: 27865514
DOI: 10.3168/jds.2016-11476 -
Frontiers in Cellular and Infection... 2022Mixed vaginitis is a complex vaginal dysbiosis that differs from single vaginitis. Vaginitis in the third trimester may lead to adverse maternal and neonatal outcomes....
Mixed vaginitis is a complex vaginal dysbiosis that differs from single vaginitis. Vaginitis in the third trimester may lead to adverse maternal and neonatal outcomes. The clinical characteristics, microbiological characteristics, and adverse pregnancy outcomes of mixed vaginitis in late pregnancy are worth studying. Therefore, this study investigated the clinical and microbiological characteristics of vaginitis and adverse pregnancy outcomes of patients with mixed vaginitis. We studied 1,674 women in late pregnancy who attended the Tianjin Medical University General Hospital from November, 2019 to October, 2021. We administered standardized questionnaires, performed vaginal examination and sampling plus microscope examinations, and assessed follow-up pregnancy outcomes. We cultured the vaginal discharge of the patients with mixed vaginitis to isolate pathogens and performed antimicrobial susceptibility tests of the isolated pathogens. For the patients with peripartum infection, we collected a sample to isolate pathogens. Among the 1,674 women, 66 (3.9%) had mixed vaginitis. The independent risk factor for mixed vaginitis in late pregnancy was a history of vaginitis during early and middle pregnancy (OR = 5.637, 95% CI: 3.314-9.580). The signs of vaginal erythema (63.6% 42.0%), yellow discharge (81.8% . 59.6%), and malodor (31.8% . 18.8%) (0.05) were significantly higher in patients with mixed vaginitis than in patients with single vaginitis. Bacterial isolates of the vaginal secretions of patients with mixed bacterial vaginitis were mainly the pathogens of aerobic vaginitis and bacterial vaginosis, such as , , and Pathogen isolation of the vaginal secretions of patients with mixed fungus and bacteria vaginitis mainly included , followed by , , , , and . Women with mixed vaginitis had an increased incidence and risk of peripartum infections (6.1% . 1.4%, <0.05; OR = 3.985, 95% CI:1.214-13.079). is the main pathogen that causes peripartum infection. Mixed vaginitis in late pregnancy is characterized by a severe and complex phenotype, complex vaginal dysbiosis, and a long course of vaginal dysbiosis. This can lead to an increased incidence and risk of peripartum infection. Therefore, more attention should be paid to patients with mixed vaginitis in the third trimester of pregnancy.
Topics: Cross-Sectional Studies; Dysbiosis; Escherichia coli; Female; Humans; Male; Pregnancy; Pregnancy Outcome; Pregnancy Trimester, Third; Vaginitis; Vaginosis, Bacterial; Vulvovaginitis
PubMed: 35419297
DOI: 10.3389/fcimb.2022.798738 -
Journal of Dairy Science Jul 2017Non-aureus staphylococci (NAS), the microorganisms most frequently isolated from bovine milk worldwide, are a heterogeneous group of numerous species. To establish their...
Non-aureus staphylococci (NAS), the microorganisms most frequently isolated from bovine milk worldwide, are a heterogeneous group of numerous species. To establish their importance as a group, the distribution of individual species needs to be determined. In the present study, NAS intramammary infection (IMI) was defined as a milk sample containing ≥1,000 cfu/mL in pure or mixed culture that was obtained from a cohort of cows assembled by the Canadian Bovine Mastitis Research Network. Overall, 6,213 (6.3%) of 98,233 quarter-milk samples from 5,149 cows and 20,305 udder quarters were associated with an NAS IMI. Of the 6,213 phenotypically identified NAS isolates, 5,509 (89%) were stored by the Canadian Bovine Mastitis Research Network Mastitis Pathogen Collection and characterized using partial sequencing of the rpoB housekeeping gene, confirming 5,434 isolates as NAS. Prevalence of each NAS species IMI was estimated using Bayesian models, with presence of a specific NAS species as the outcome. Overall quarter-level NAS IMI prevalence was 26%. The most prevalent species causing IMI were Staphylococcus chromogenes (13%), Staphylococcus simulans (4%), Staphylococcus haemolyticus (3%), Staphylococcus xylosus (2%), and Staphylococcus epidermidis (1%). The prevalence of NAS IMI as a group was highest in first-parity heifers and was evenly distributed throughout cows in parities ≥2. The IMI prevalence of some species such as S. chromogenes, S. simulans, and S. epidermidis differed among parities. Overall prevalence of NAS IMI was 35% at calving, decreased over the next 10 d, and then gradually increased until the end of lactation. The prevalence of S. chromogenes, Staphylococcus gallinarum, Staphylococcus cohnii, and Staphylococcus capitis was highest at calving, whereas the prevalence of S. chromogenes, S. haemolyticus, S. xylosus, and S. cohnii increased during lactation. Although the overall prevalence of NAS IMI was similar across barn types, the prevalence of S. simulans, S. xylosus, S. cohnii, Staphylococcus saprophyticus, S. capitis, and Staphylococcus arlettae IMI was higher in tiestall barns; the prevalence of S. epidermidis IMI was lowest; and the prevalence of S. chromogenes and Staphylococcus sciuri IMI was highest in bedded-pack barns. Staphylococcus simulans, S. epidermidis, S. xylosus, and S. cohnii IMI were more prevalent in herds with intermediate to high bulk milk somatic cell count (BMSCC) and S. haemolyticus IMI was more prevalent in herds with high BMSCC, whereas other common NAS species IMI were equally prevalent in all 3 BMSCC categories. Distribution of NAS species IMI differed among the 4 regions of Canada. In conclusion, distribution differed considerably among NAS species IMI; therefore, accurate identification (species level) is essential for studying NAS epidemiology.
Topics: Animals; Bayes Theorem; Canada; Cattle; Cattle Diseases; Female; Mammary Glands, Animal; Mastitis, Bovine; Milk; Pregnancy; Prevalence; Species Specificity; Staphylococcal Infections; Staphylococcus
PubMed: 28527793
DOI: 10.3168/jds.2016-12478 -
Research in Veterinary Science May 2023Single-layer centrifugation (SLC) with a low-density colloid is an efficient method for removing contaminating microorganisms from boar semen while recovering most...
Single-layer centrifugation (SLC) with a low-density colloid is an efficient method for removing contaminating microorganisms from boar semen while recovering most spermatozoa from the original sample. This study tested the performance of this technique, using 50-ml tubes, by spiking commercial semen doses prepared without antibiotics with selected bacterial species followed by storage at 17 °C. The doses were spiked up to 10/ml CFU (colony forming units) of the bacteria Burkholderia ambifaria, Pseudomonas aeruginosa, and Staphylococcus simulans. The semen was processed by SLC (15 ml of sample and 15 ml of colloid) with the colloid Porcicoll at 20% (P20) and 30% (P30), with a spiked control (CTL) and an unspiked control (CTL0), analyzing microbiology and sperm quality on days 0, 3 and 7. SLC completely removed B. ambifaria and S. simulans, considerably reducing P. aeruginosa and overall contamination (especially P30, ∼10 CFU/ml of total contamination on day 7, median). Sperm viability was lower in P20 and P30 samples at day 0, with higher cytoplasmic ROS. Still, results were similar in all groups on day 3 and reversed on day 7, indicating a protective effect of SLC (possibly directly by removal of damaged sperm and indirectly because of lower bacterial contamination). Sperm chromatin was affected by the treatment (lower DNA fragmentation and chromatin decondensation) and storage (higher overall condensation on day 7 as per chromomycin A3 and monobromobimane staining). In conclusion, SLC with low-density colloids can remove most bacteria in a controlled contamination design while potentially improving sperm quality and long-term storage at practical temperatures.
Topics: Male; Animals; Swine; Semen; Spermatozoa; Semen Analysis; Semen Preservation; Centrifugation; Colloids; Chromatin; Burkholderia; Sperm Motility
PubMed: 37031470
DOI: 10.1016/j.rvsc.2023.03.024