Intramammary infection with coagulase-negative staphylococci at parturition: Species-specific prevalence, risk factors, and effect on udder health.

Coagulase-negative staphylococci (CNS) are the main cause of bovine intramammary infections (IMI) in many countries. Despite a high prevalence of CNS IMI at parturition, species-specific risk factor studies, relying on accurate identification methods, are lacking. Therefore, this observational study aimed at determining the prevalence and distribution of different CNS species causing IMI in fresh heifers and dairy cows in Flemish dairy herds and identifying associated species- and subgroup-specific risk factors at the herd, cow, and quarter level. The effect on udder health was investigated as well. Staphylococcus chromogenes, S. sciuri, and S. cohnii were the most frequently isolated species. The only CNS species causing IMI in fresh heifers and dairy cows in all herds was Staphylococcus chromogenes, whereas large between-herd differences in distribution were observed for the other species. Quarters from heifers and quarters with an inverted teat end had higher odds of being infected with S. chromogenes, S. simulans, or S. xylosus as well as with S. chromogenes solely. Prepartum teat apex colonization with S. chromogenes increased the likelihood of S. chromogenes IMI in the corresponding quarters at parturition. Quarters with dirty teat apices before calving were more likely to be infected with S. cohnii, S. equorum, S. saprophyticus, or S. sciuri, supporting the environmental nature of these CNS species. Three species (S. chromogenes, S. simulans, and S. xylosus) were associated with a higher quarter somatic cell count at parturition as compared with uninfected quarters.

De Visscher A ，Piepers S ，Haesebrouck F ，De Vliegher S ... -  《-》

Short communication: Species group-specific predictors at the cow and quarter level for intramammary infection with coagulase-negative staphylococci in dairy cattle throughout lactation.

Coagulase-negative staphylococci (CNS) are frequently isolated from quarters with subclinical mastitis, teat apices, and the cows' environment. Virulence, ecology, epidemiological behavior, and effect on udder health vary between different CNS species. Staphylococcus chromogenes, Staph. simulans, and Staph. xylosus are frequently present in milk and have a more substantial effect on quarter milk somatic cell count than other species. Therefore, these species are considered the "more relevant" CNS. As species-specific factors associated with CNS intramammary infection (IMI) have not yet been identified and susceptibility for IMI differs between cows and quarters, this study aimed to identify predictors for CNS IMI at the cow and quarter level (some of them changing over time) with a specific focus on the aforementioned more relevant CNS. Precise data were available from a longitudinal study (3,052 observations from 344 quarters from 86 dairy cows belonging to 3 commercial dairy herds). All CNS were molecularly identified to the species level, and multivariable, multilevel logistic regression models taking into account the longitudinal nature of the data, were fit to study the likelihood of infection. Staphylococcus chromogenes, Staph. xylosus, and Staph. cohnii were the most frequently isolated species from CNS IMI in older cows, whereas Staph. chromogenes, Staph. xylosus, and Staph. simulans were the main species found in IMI in heifers. Quarters from heifers (as opposed to multiparous cows), from heifers and multiparous cows in third or fourth month in lactation (as opposed to early lactation, <60 d in milk), and with an increasing quarter milk SCC were more likely to be infected with the more relevant CNS species. Quarter milk SCC was identified as the sole statistically significant predictor for IMI with other CNS species, although the size of the effect was lower [odds ratio of 1.6 (1.4-1.9) vs. 2.1 (1.8-2.5)] than the effect for IMI with the more relevant CNS. As a strong herd effect was present, studying herd-level predictors is warranted.

De Visscher A ，Piepers S ，Supré K ，Haesebrouck F ，De Vliegher S ... -  《-》

Prevalence of non-aureus staphylococci species causing intramammary infections in Canadian dairy herds.

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.

Condas LAZ ，De Buck J ，Nobrega DB ，Carson DA ，Naushad S ，De Vliegher S ，Zadoks RN ，Middleton JR ，Dufour S ，Kastelic JP ，Barkema HW ... -  《-》

Teat apex colonization with coagulase-negative Staphylococcus species before parturition: Distribution and species-specific risk factors.

Coagulase-negative staphylococci (CNS) are the main cause of bovine intramammary infections and are also abundantly present in extramammary habitats such as teat apices. Teat apex colonization (TAC) with CNS has already been explored in lactating dairy cows at the species level, whereas this is not true for dry cows and end-term heifers. Therefore, the aim of this observational study was to describe CNS TAC in nonlactating dairy cows and end-term heifers in Flemish dairy herds and to identify associated risk factors at the herd, cow, and quarter level. All CNS were molecularly identified to the species level using transfer RNA intergenic spacer PCR (tDNA-PCR) and sequencing of the 16S rRNA gene, allowing for species-specific statistical analyses using multivariable, multilevel logistic regression. Staphylococcus devriesei, Staphylococcus chromogenes, Staphylococcus haemolyticus, and Staphylococcus equorum were the most frequently isolated species. Staphylococcus chromogenes was the sole species colonizing teat apices of cows and heifers in all herds, whereas large between-herd differences were observed for the other species. Teat apices of red and white Holstein Friesians, of quarters dried off without an internal teat sealer, and swabbed in months with lower precipitation and higher ambient temperature were significantly more likely to be colonized by S. devriesei. Slightly dirty teat apices and teat apices swabbed in months with lower precipitation had higher odds of being colonized by S. chromogenes, whereas teat apices sampled in months with lower precipitation and higher ambient temperature were more likely to be colonized by S. haemolyticus. Dirty teat apices and teat apices swabbed in months with lower ambient temperature in combination with low precipitation had higher odds of being colonized by S. equorum. Diverse factors explaining CNS TAC, yet mostly related to humidity, ambient temperature, and hygiene, substantiate differences in epidemiological behavior and ecology between species.

De Visscher A ，Piepers S ，Haesebrouck F ，De Vliegher S ... -  《-》

Longitudinal study on the effects of intramammary infection with non-aureus staphylococci on udder health and milk production in dairy heifers.

We conducted a longitudinal study to evaluate the effect of non-aureus staphylococci (NAS) causing subclinical intramammary infections (IMI) on quarter milk somatic cell count (qSCC) and quarter milk yield (qMY). In total, 324 quarters of 82 Holstein Friesian heifers were followed from calving to 130 d in milk (DIM) and were sampled 10 times each at 14-d intervals. The IMI status of each quarter was determined based on bacterial culture results at the current and previous or next sampling day, or both. The qSCC was determined on each sampling day and the average qMY on sampling day was available through stored daily milk weight data in the management program of the automatic milking system. A transient IMI (tIMI) was defined as a case where a specific pathogen was isolated from a quarter on only one sampling day and not on the previous or next sampling day. When the same bacterial strain, as defined by random amplification of polymorphic DNA-PCR, was isolated from the same quarter on multiple sampling days, it was defined as a persistent IMI (pIMI) status on those sampling days; a pIMI episode was defined as the combination of multiple consecutive pIMI statuses with the same bacterial strain on different sampling days. During this study, 142 subclinical IMI with NAS occurred in 116 different quarters from 64 animals, yielding in total 304 NAS isolates belonging to 17 different species. The prevalence of NAS was highest in the first 4 DIM. Overall, the predominant species was Staphylococcus chromogenes (52% of the isolates), followed by S. epidermidis (9.2%), S. xylosus (8.2%), and S. equorum (5.9%). Staphylococcus chromogenes was the only species for which an effect on qSCC and qMY could be analyzed separately; the other NAS species were considered as a group because of their low prevalence. Eighteen out of 40 IMI (45%) caused by S. chromogenes persisted over at least 2 sampling days, whereas only 10 of 102 (9.8%) IMI caused by other NAS species persisted for at least 2 sampling days. The average duration of pIMI episodes was 110.4 d for S. chromogenes and 70 d for the other NAS species. Remarkably, 17 of the 18 pIMI episodes with S. chromogenes started within the first 18 DIM. The qSCC was highest in quarters having a pIMI with a major pathogen, followed by quarters having a pIMI with S. chromogenes, and a pIMI with other NAS. Transient IMI with other NAS or with a major pathogen caused a small but significantly higher qSCC, whereas the qSCC in quarters having a tIMI with S. chromogenes was not statistically different compared with noninfected quarters. No significant differences in qMY were observed between quarters having a pIMI or tIMI with S. chromogenes or with the other NAS species compared with noninfected quarters, despite the higher qSCC. Quarters having a pIMI with major pathogens showed significantly lower daily milk production. Surprisingly, quarters that cured from an IMI with S. chromogenes had a significantly lower qMY than noninfected quarters.

Valckenier D ，Piepers S ，Schukken YH ，De Visscher A ，Boyen F ，Haesebrouck F ，De Vliegher S ... -  《-》