Shedding routes of Coxiella burnetii in dairy cows: implications for detection and control

Reliable detection of Coxiella burnetii shedders is a critical point for the control of the spread of this bacterium among animals and from animals to humans. Coxiella burnetii is shed by ruminants mainly by birth products (placenta, birth fluids), but may also be shed by vaginal mucus, milk, and faeces, urine and semen. However, the informative value of these types of samples to identify shedders under field conditions is unknown. Our aim was then to describe the responses obtained using a real-time PCR technique applied to milk, vaginal mucus and faeces samples taken from 242 dairy cows in commercial dairy herds known to be naturally infected with Coxiella burnetii, and to assess their putative associations. Positive results were found in all types of tested samples even in faeces. No predominant shedding route was identified. Among the shedder cows, 65.4% were detected as shedders by only one route. By contrast, cows with positive results for all three samples were scarce (less than 7%). Testing a cow based on only one type of biological sample may lead to misclassify it with regards to its shedding of Coxiella burnetii and thereby underestimate the risk of bacterial spread within a herd.     

Shedding and serological patterns of dairy cows following abortions associated with Coxiella burnetii DNA detection

To describe both shedding and serological patterns following abortions detected as being associated with Coxiella burnetii (Cb), 24 cows experiencing an abortion due to Cb were followed over a one month period. Samples taken on the day of abortion (D0) were followed 3-fold by weekly samplings from day 14 (D14) to D28 after the abortion. Milk and vaginal mucus were collected at each weekly sampling and tested using real-time PCR while blood samples were collected 2-fold on D21 and D28 and tested using ELISA. We found a very short duration of C. burnetii shedding in vaginal mucus after abortion, highlighting the need to collect samples as rapidly as possible following an abortion to avoid false negative results. In contrast with previous results, concomitancy of vaginal and mucus shedding was frequent, especially for cows shedding a high bacterial load on DO leading to the hypothesis that the clinical onset of the infection influences the modalities of Cb shedding. Lastly, serological results indicating a lack of sensitivity to detect Cb shedder cows (especially for cows for which Ct values were high) suggest that ELISA is not a useful tool to diagnose abortions at the individual level.     

Effectiveness of vaccination and antibiotics to control Coxiella burnetii shedding around calving in dairy cows

Effectiveness of phase 1 vaccine, combined or not with tetracycline, to control Coxiella burnetii vaginal shedding at calving in cows was assessed through a 13 months study in 22 Q fever clinically affected commercial dairy herds. Four medical strategies implemented at herd level but randomly assigned to cows (vaccination, vaccination and tetracycline, tetracycline, nothing) were compared. There was no significant interaction effect between vaccination and antibiotherapy. Tetracycline used once at drying off was associated with a lower risk of being detected shedder at calving (OR=0.40, CI 95% [0.21-0.75]), but had no significant effect on the bacterial load shed. Vaccination did not significantly prevent shedding but was significantly (OR=0.15, CI 95% [0.03-0.85]) associated with lower bacterial load shed. Thus, vaccination using a phase 1 vaccine and antibiotherapy using tetracycline is associated with a decrease in shedding in dairy cows and could contribute to reduce the bacterial load generated in the environment. To the best of our knowledge, this is the first study providing useful information for evidenced-based and rational use of medical strategy combining antibiotic and vaccination in infected dairy cattle herds.     

Beyond occurrence: Body condition and stress hormone as integrative indicators of habitat availability and fragmentation in the common toad

Availability and fragmentation of habitat are today identified as key determinants of population maintenance. Disentangling their relative effects is of crucial importance to adequately manage landscapes. However, the classical use of occurrence or abundance data as proxy to assess the responses of populations could be insufficient to detect less drastic impacts than short-term population extinction. We evaluated the effects of both habitat availability and fragmentation at three spatial scales on both the occurrence and two “physiological state” indicators (body condition and level of stress hormone) reflecting the physical state of common toads (Bufo bufo). Our study showed that occurrence was negatively affected only by habitat availability at the largest spatial scale whereas the both physiological state indicators studied at the population scale are significantly altered by both habitat availability and fragmentation at the finest spatial scale. Results obtained with occurrence and physiological state approaches substantially diverge, which highlights the need to also investigate proximal processes to deeply understand how populations are threatened by landscape modifications. The use of physiological state indicators, particularly of body condition, powerful and easy to obtain, could be particularly relevant to detect early warnings of population decline allowing a management before extinction. Only a complete investigation of the potential impacts of landscape on the different population processes could offer a realistic picture of the requirements of populations to avoid drastic alterations.     

Reduced Atmospheric Manganese in Montreal Following Removal of Methylcyclopentadienyl Manganese Tricarbonyl (MMT)

Methylcyclopentadienyl manganese tricarbonyl (MMT) was used between 1990 and 2003 as an antiknock agent and as an octane booster in Canadian unleaded gasoline. Its combustion leads to Mn emissions. The objective of this research was to examine the variations in atmospheric Mn in Montreal (Canada) from 2001 to 2007, covering the period prior (2001?C2003) to and following (2005?C2007) MMT use. Three sampling stations were selected because of their proximity to roads with widely differing and well-known traffic patterns. Filters from 2001 to 2007 were obtained from the Montreal Urban Community. The first sample of each month was selected, and Mn analysis was performed by neutron activation analysis. Total suspended particulates (TSP) was calculated by weighing the filters before and after dust collection. Results show a significant decrease of Mn over time at each station, whereas TSP decreased significantly in two stations. Comparing atmospheric Mn during and after the period of use of MMT 2001?C2003 vs 2005?C2007 showed a significant decrease at all stations. For TSP, only one station showed borderline significant decline between these two periods. Overall, between the two periods, Mn and TSP decreased by 39% and 17%, respectively. These data suggest that the combustion of MMT led to an increase of airborne Mn of approximately 22%. These findings should help in decision-making processes concerning the use of MMT in gasoline in other countries.     

Anaesthetic efficacy of clove oil,benzocaine, 2‐phenoxyethanol and tricaine methanesulfonate in juvenile marbled spinefoot (Siganus rivulatus)

Anaesthetics are used in aquaculture and fisheries to facilitate routine procedures, such as capture, handling, transportation, tagging, grading and measurements that can often cause injury or induce physiological stress. Two experiments were performed to assess the efficacies of four anaesthetic agents, clove oil, benzocaine, 2‐phenoxyethanol and MS‐222 on juvenile marbled spinefoot rabbitfish (Siganus rivulatus). In the first experiment we tested the lowest effective doses that produced induction and recovery times in 3 min or less and 5 min or less respectively. Dosages were 70 mg L^?1 for clove oil, 60–70 mg L^?1 for benzocaine, 400 μL L^?1 for 2‐phenoxyethanol and 100–125 mg L^?1 for MS‐222. In the second experiment, we determined optimal concentrations of the four anaesthetics if they were to be used to transport rabbitfish fry. Anaesthetic concentrations suitable for handling and transport were: 10–15 mg L^?1 of MS‐222, 5–10 mg L^?1 of benzocaine, 5 mg L^?1 of clove oil and 50–100 μL L^?1 of 2‐phenoxyethanol. All anaesthetic agents are acceptable for use on S. rivulatus, however, 2‐phenoxyethanol, MS‐222 and clove oil appear to be more suitable than benzocaine. Further studies need to be conducted on effects of high and low doses of anaesthetic agents on physiology of marbled spinefoot.     

Clove oil as an anaesthetic for Australian redclaw crayfish Cherax quadricarinatus

Crustaceans are aquacultured both for food and as ornamental organisms. Pain and distress are not entirely understood in crustaceans, but the industry is moving towards requiring protection for the welfare of these animals during handling. In the present study, we evaluated the efficacy of clove oil as an anaesthetic for redclaw crayfish (Cherax quadricarinatus) as a model for freshwater crustaceans. We also studied how factors such as body weight and sex of redclaw could affect this efficacy. The whole experiment was replicated in two consecutive years. Redclaw juveniles were sorted into three size classes: small (<5 g), medium (5–12 g) and large (12–37 g). At least 10 males and 10 females from each size class were placed individually in water containing clove oil concentrations of 375 and 500 μl/L. Both concentrations induced rapid induction and recovery times, with 500 μl/L being the more effective concentration of the two. Induction and recovery times increased with the increase in crayfish size. No significant differences were found in induction and recovery times between male and female crayfish. Results suggest that clove oil is an effective anaesthetic for redclaw.