S.M. Andrew

Indication of a Lombard vocal response in the St. Lawrence River beluga

Noise pollution is recognized as a potential danger to marine mammals in general, and to the St. Lawrence beluga in particular. One method of determining the impacts of noise on an animals communication is to observe a natural and repeatable response of the vocal system to variations in noise level. This is accomplished by observing intensity changes in animal vocalizations in response to environmental noise. One such response observed in humans, songbirds, and some primates is the Lombard vocal response. This response represents a vocal system reaction manifested by changes in vocalization level in direct response to changes in the noise field. In this research, a population of belugas in the St. Lawrence River Estuary was tested to determine whether a Lombard response existed by using hidden Markhov-classified vocalizations as targets for acoustical analyses. Correlation and regression analyses of signals and noise indicated that the phenomenon does exist. Further, results of human subjects experiments [Egan, J. J. (1966), Ph.D. dissertation; Scheifele, P. M. (2003), Ph.D. dissertation], along with previously reported data from other animal species, are similar to those exhibited by the belugas. Overall, findings suggest that typical noise levels in the St. Lawrence River Estuary have a detectable effect on the communication of the beluga.

Diverse Antibiotic Resistance Genes in Dairy Cow Manure

ABSTRACT Application of manure from antibiotic-treated animals to crops facilitates the dissemination of antibiotic resistance determinants into the environment. However, our knowledge of the identity, diversity, and patterns of distribution of these antibiotic resistance determinants remains limited. We used a new combination of methods to examine the resistome of dairy cow manure, a common soil amendment. Metagenomic libraries constructed with DNA extracted from manure were screened for resistance to beta-lactams, phenicols, aminoglycosides, and tetracyclines. Functional screening of fosmid and small-insert libraries identified 80 different antibiotic resistance genes whose deduced protein sequences were on average 50 to 60% identical to sequences deposited in GenBank. The resistance genes were frequently found in clusters and originated from a taxonomically diverse set of species, suggesting that some microorganisms in manure harbor multiple resistance genes. Furthermore, amid the great genetic diversity in manure, we discovered a novel clade of chloramphenicol acetyltransferases. Our study combined functional metagenomics with third-generation PacBio sequencing to significantly extend the roster of functional antibiotic resistance genes found in animal gut bacteria, providing a particularly broad resource for understanding the origins and dispersal of antibiotic resistance genes in agriculture and clinical settings. IMPORTANCE The increasing prevalence of antibiotic resistance among bacteria is one of the most intractable challenges in 21st-century public health. The origins of resistance are complex, and a better understanding of the impacts of antibiotics used on farms would produce a more robust platform for public policy. Microbiomes of farm animals are reservoirs of antibiotic resistance genes, which may affect distribution of antibiotic resistance genes in human pathogens. Previous studies have focused on antibiotic resistance genes in manures of animals subjected to intensive antibiotic use, such as pigs and chickens. Cow manure has received less attention, although it is commonly used in crop production. Here, we report the discovery of novel and diverse antibiotic resistance genes in the cow microbiome, demonstrating that it is a significant reservoir of antibiotic resistance genes. The genomic resource presented here lays the groundwork for understanding the dispersal of antibiotic resistance from the agroecosystem to other settings. The increasing prevalence of antibiotic resistance among bacteria is one of the most intractable challenges in 21st-century public health. The origins of resistance are complex, and a better understanding of the impacts of antibiotics used on farms would produce a more robust platform for public policy. Microbiomes of farm animals are reservoirs of antibiotic resistance genes, which may affect distribution of antibiotic resistance genes in human pathogens. Previous studies have focused on antibiotic resistance genes in manures of animals subjected to intensive antibiotic use, such as pigs and chickens. Cow manure has received less attention, although it is commonly used in crop production. Here, we report the discovery of novel and diverse antibiotic resistance genes in the cow microbiome, demonstrating that it is a significant reservoir of antibiotic resistance genes. The genomic resource presented here lays the groundwork for understanding the dispersal of antibiotic resistance from the agroecosystem to other settings.

Antibacterial effect of plant-derived antimicrobials on major bacterial mastitis pathogens in vitro.

The objective of this study was to investigate the antimicrobial effect of plant-derived antimicrobials including trans-cinnamaldehyde (TC), eugenol, carvacrol, and thymol on major bacterial mastitis pathogens in milk. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the aforementioned compounds on Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, Staphylococcus aureus, and Escherichia coli were determined. In addition, the bactericidal kinetics of TC on the aforementioned pathogens and the persistence of the antimicrobial activity of TC in milk over a period of 2 wk were investigated. All 4 plant-derived molecules exhibited antimicrobial activity against the 5 mastitis pathogens tested, but TC was most effective in killing the bacteria. The MIC and MBC of TC on Staph. aureus, E. coli, and Strep. uberis were 0.1 and 0.45%, respectively, whereas that on Strep. agalactiae and Strep. dysgalactiae were 0.05 and 0.4%, respectively. The MIC and MBC of the other 3 molecules ranged from 0.4 to 0.8% and 0.8 to 1.5%, respectively. In time-kill assays, TC at the MBC reduced the bacterial pathogens in milk by 4.0 to 5.0 log(10) cfu/mL and to undetectable levels within 12 and 24 h, respectively. The antimicrobial effect of TC persisted for the duration of the experiment (14 d) without any loss of activity. Results of this study suggest that TC has the potential to be evaluated as an alternative or adjunct to antibiotics as intramammary infusion to treat bovine mastitis.

Factors associated with the risk of antibiotic residues and intramammary pathogen presence in milk from heifers administered prepartum intramammary antibiotic therapy.

Heifers (n=136) from 5 herds were treated with a commercially available beta-lactam intramammary (IMM) antibiotic preparation containing cephapirin sodium at 10-21 d prior to anticipated parturition to evaluate the risk of antibiotic residues occurring in milk postpartum and to determine factors associated with antibiotic residues and IMM pathogen presence in milk postpartum. Mammary secretions collected from quarters before antibiotic administration and during weeks 1, 2 and 3 postpartum were analyzed for mastitis pathogens. Composite milk was collected at milkings 3, 6 and 10 postpartum and analyzed for beta-lactam residues using a microbial inhibition antibiotic residue screening test. Antibiotic residues were confirmed with beta-lactamase treatment and re-tested for residues. Residues were detected in 28.0, 8.82 and 3.68% of milk samples obtained at the third, sixth, and tenth milking postpartum, respectively. Increases in interval between prepartum antibiotic therapy and parturition and an increase in the postpartum interval to sampling were associated with a decrease in risk of antibiotic residues. The presence of antibiotic residues in milk at the third milking was associated with a reduced risk for IMM pathogen prevalence in the first 21 d postpartum. Lower somatic cell counts, an increase in mean milk yield over 200 days in milk and a reduction in IMM pathogen prevalence were associated with the presence of an antibiotic in milk postpartum. Screening milk for antibiotic residues in milk postpartum following prepartum antibiotic therapy in heifers is recommended to reduce the risk for antibiotic residue contamination of milk.

Evaluation of a clay-based acidic bedding conditioner for dairy cattle bedding

This study investigated the effects of a clay-based acidic bedding conditioner on sawdust bedding pH, dry matter (DM), environmental pathogen counts, and environmental bacterial counts on teat ends of lactating dairy cows. Sixteen lactating Holstein cows were paired based on parity, days in milk, milk yield, and milk somatic cell count, and were negative for the presence of an intramammary pathogen. Within each pair, cows were randomly assigned to 1 of 2 treatments with 3-wk periods in a crossover design. Treatment groups consisted of 9 freestalls per group bedded with either untreated sawdust or sawdust with a clay-based acidic bedding conditioner, added at 3- to 4-d intervals over each 21-d period. Bedding and teat ends were aseptically sampled on d 0, 1, 2, 7, 14, and 21 for determination of environmental bacterial counts. At the same time points, bedding was sampled for DM and pH determination. The bacteria identified in the bedding material were total gram-negative bacteria, Streptococcus spp., and coliform bacteria. The bacteria identified on the teat ends were Streptococcus spp., coliform bacteria, and Klebsiella spp. Teat end score, milk somatic cell count, and intramammary pathogen presence were measured weekly. Bedding and teat cleanliness, environmental high and low temperatures, and dew point data were collected daily. The bedding conditioner reduced the pH, but not the DM, of the sawdust bedding compared with untreated sawdust. Overall environmental bacterial counts in bedding were lower for treated sawdust. Total bacterial counts in bedding and on teat ends increased with time over both periods. Compared with untreated sawdust, the treated bedding had lower counts of total gram-negative bacteria and streptococci, but not coliform counts. Teat end bacterial counts were lower for cows bedded on treated sawdust for streptococci, coliforms, and Klebsiella spp. compared with cows bedded on untreated sawdust. The clay-based acidic bedding conditioner reduced environmental pathogens in sawdust bedding and teat ends without affecting teat end integrity.