Ketoprofen pharmacokinetics of R‐ and S‐isomers in juvenile loggerhead sea turtles (Caretta caretta) after single intravenous and single‐ and multidose intramuscular administration

Ketoprofen is a nonsteroidal anti‐inflammatory and analgesic agent that nonselectively inhibits cyclooxygenase, with both COX‐1 and COX‐2 inhibition. Recent studies on COX receptor expression in reptiles suggest that nonselective COX inhibitors may be more appropriate than more selective inhibitors in some reptiles, but few pharmacokinetic studies are available. The goal of this study was to determine single‐ and multidose (three consecutive days) pharmacokinetics of racemic ketoprofen administered intravenously and intramuscularly at 2 mg/kg in healthy juvenile loggerhead turtles (Caretta caretta). The S‐isomer is the predominant isomer in loggerhead sea turtles, similar to most mammals, despite administration of a 50:50 racemic mixture. Multidose ketoprofen administration demonstrated no bioaccumulation; therefore, once‐daily dosing will not require dose adjustment over time. S‐isomer pharmacokinetic parameters determined in this study were C_max of 10.1 μg/ml by IM injection, C₀ of 13.4 μg/ml by IV injection, AUC of 44.7 or 69.4 μg*hr/ml by IM or IV injection, respectively, and T½ of 2.8 or 3.6 hr by IM or IV injection, respectively. Total ketoprofen plasma concentrations were maintained for at least 12 hr above concentrations determined to be effective for rats and humans. A dose of 2 mg/kg either IM or IV every 24 hr is likely appropriate for loggerhead turtles.     

A repeated cross‐sectional study of the epidemiology of Campylobacter and antimicrobial resistant Enterobacteriaceae in free‐living Canada geese in Guelph,Ontario, Canada

From May through October 2016, we conducted a repeated cross‐sectional study examining the effects of temporal, spatial, flock and demographic factors (i.e. juvenile vs. adult) on the prevalence of Campylobacter and antimicrobial resistant Enterobacteriaceae among 344 fresh faecal samples collected from Canada geese (Branta canadensis) from four locations where birds nested in Guelph, Ontario, Canada. The overall prevalence of Campylobacter among all fresh faecal samples was 9.3% and was greatest in the fall when these birds became more mobile following the nesting season. Based on 40 gene comparative genomic fingerprinting (CGF40), the increase in prevalence noted in the fall was matched by an increase in the number of unique CGF40 subtypes identified. Resistance to colistin was detected most commonly, in 6% of Escherichia coli isolates, and was highest in the late summer months. All colistin‐resistant isolates were negative for the mcr‐1 to mcr‐5 genes; a chromosomal resistance mechanism (PmrB) was identified in all of these isolates. The prevalence of samples with E. coli exhibiting multi‐class resistance or extended spectrum beta‐lactamase was low (i.e. <2% of samples). The intra‐class correlation coefficients, estimated from the variance components of multilevel logistic regression models, indicated that the shedding of Campylobacter and antimicrobial resistant E. coli among geese within a flock (i.e. birds collected from the same site on the same day) was moderately correlated. Spatial, temporal, and spatiotemporal clusters identified using the spatial scan statistic, largely supported the findings from our multi‐level models. Salmonella was not isolated from any of the fresh faecal samples collected suggesting that its prevalence in this population of birds was very low.     

The occurrence of Salmonella,extended‐spectrum β‐lactamase producing Escherichia coli and carbapenem resistant non‐fermenting Gram‐negative bacteria in a backyard poultry flock environment

Increase in the number of small‐scale backyard poultry flocks in the USA has substantially increased human‐to‐live poultry contact, leading to increased public health risks of the transmission of multi‐drug resistant (MDR) zoonotic and food‐borne bacteria. The objective of this study was to detect the occurrence of Salmonella and MDR Gram‐negative bacteria (GNB) in the backyard poultry flock environment. A total of 34 backyard poultry flocks in Washington State (WA) were sampled. From each flock, one composite coop sample and three drag swabs from nest floor, waterer‐feeder, and a random site with visible faecal smearing, respectively, were collected. The samples were processed for isolation of Salmonella and other fermenting and non‐fermenting GNB under ceftiofur selection. Each isolate was identified to species level using MALDI‐TOFF and tested for resistance against 16 antibiotics belonging to eight antibiotic classes. Salmonella serovar 1,4,[5],12:i:‐ was isolated from one (3%) out of 34 flocks. Additionally, a total of 133 ceftiofur resistant (Cef^R) GNB including Escherichia coli (53), Acinetobacter spp. (45), Pseudomonas spp. (22), Achromobacter spp. (8), Bordetella trematum (1), Hafnia alvei (1), Ochrobactrum intermedium (1), Raoultella ornithinolytica (1), and Stenotrophomonas maltophilia (1) were isolated. Of these, 110 (82%) isolates displayed MDR. Each flock was found positive for the presence of one or more Cef^R GNB. Several MDR E. coli (n = 15) were identified as extended‐spectrum β‐lactamase (ESBL) positive. Carbapenem resistance was detected in non‐fermenting GNB including Acinetobacter spp. (n = 20), Pseudomonas spp. (n = 11) and Stenotrophomonas maltophila (n = 1). ESBL positive E. coli and carbapenem resistant non‐fermenting GNB are widespread in the backyard poultry flock environment in WA State. These GNB are known to cause opportunistic infections, especially in immunocompromised hosts. Better understanding of the ecology and epidemiology of these GNB in the backyard poultry flock settings is needed to identify potential risks of transmission to people in proximity.     

Genetic background of CTX‐M‐15‐producing Enterobacter hormaechei ST114 and Citrobacter freundii ST265 co‐infecting a free‐living green turtle (Chelonia mydas)

CTX‐M‐type extended‐spectrum β‐lactamase (ESBL)‐producing Enterobacteriaceae have become identified in marine ecosystem constituting a serious ecological issue. In this respect, although contamination of coastal waters and seafood, and even colonization of seabirds and fishes have been increasingly reported, molecular data are lacking to elucidate the clinical impact of ESBL producers in infected marine animals. In this study, using a genomic approach, we have analysed the genetic background of CTX‐M‐15‐producing Enterobacter hormaechei (belonging to the international human clone ST114) and Citrobacter freundii (ST265) co‐infecting a free‐living green turtle (Chelonia mydas) suffering from septic arthritis, which progressed to generalized coelomitis and death. Wide resistome of these pathogens contributed to treatment failure and death of the animal.     

Effect of antimicrobial use and production system on Campylobacter spp., Staphylococcus spp. and Salmonella spp. resistance in Spanish swine: A cross‐sectional study

Antimicrobial resistance is a worldwide public health threat; hence, current trends tend to reduce antimicrobial use in food‐producing animals and to monitor resistance in primary production. This study aimed at evaluating the impact of antimicrobial use and production system on swine farms in the antimicrobial resistance of Campylobacter, Salmonella and Staphylococcus, the main zoonotic pathogens in pig herds, in order to assess their potential value as sentinel microorganisms in antimicrobial resistance surveillance schemes. A total of 37 Spanish swine farms, 18 intensive and 19 organic/extensive farms, were included in the study. The antimicrobial resistance of 104 Campylobacter, 84 Staphylococcus and 17 Salmonella isolates was evaluated using Sensititre plates following the EUCAST guidelines. Mixed‐effects logistic regression was used to evaluate the influence of production system and antimicrobial use in resistant and multidrug‐resistant (MDR) phenotypes to the antimicrobials tested. The results showed that antimicrobial use was higher (p < .001) on intensive farms than on organic/extensive farms. MDR in Campylobacter and Staphylococcus was lower on organic/extensive farms (OR < .01p < .001). Antimicrobial resistance in Campylobacter and Staphylococcus isolates was, also for most of the antimicrobials studied, significantly higher in intensive than organic/extensive pig herds. Tetracycline resistance was associated with total antimicrobial consumption in both microbial species (p < .05), and some cross‐associations between distinct antimicrobial substances were established, for instance resistance to erythromycin was associated with macrolide and phenicol consumption. No significant associations could be established for Salmonella isolates. The results demonstrate the link between antimicrobial consumption and resistance in zoonotic bacteria and evidence the potential value of using Campylobacter and Staphylococcus species in monitoring activities aimed at determining the impact of antimicrobials use/reduction on the occurrence and spread of antimicrobial resistance.     

Molecular Characterization and Serology of Leptospira kirschneri (Serogroup Grippotyphosa) Isolated from Urine of a Mare Post‐Abortion in Brazil

A strain of Leptospira kirschneri (serogroup Grippotyphosa) was cultured from urine of a mare post‐abortion in Brazil and characterized by serogrouping, multiple‐locus variable‐number tandem repeat analysis, PGFE, and sequencing of genes rrs and secY. Strains of L. kirschneri have apparently never been recovered from horses in tropical area, only in Europe and USA. Knowledge of local epidemiology is important to interpret genetic profiles of leptospires circulating in an area.     

Comparison of plasma vitamin A and E,copper and zinc levels in free‐ranging and captive greater flamingos (Phoenicopterus roseus) and their relation to pododermatitis

Pododermatitis is a worldwide problem in captive flamingos. Studies in domestic poultry showed that nutrition is a possible influencing factor for pododermatitis. Vitamin A and E, copper and zinc levels were analysed in two different diets (diet 1 = in‐house mix and diet 2 = commercial diet) and in plasma of captive greater flamingos fed these diets and compared to those of free‐ranging greater flamingos. Results were analysed with respect to type and severity of foot lesions of the individuals from the different groups. Juvenile and subadult/adult captive flamingos on diet 1 showed various types and severities of foot lesions, whereas no foot lesions were found at the time of blood sampling in juvenile captive flamingos on diet 2. Juvenile captive flamingos on diet 1 had significantly lower plasma zinc levels than juvenile captive flamingos on diet 2 and juvenile free‐ranging flamingos; data were also lower than reference ranges for flamingos, poultry and cranes. There were no significant differences in plasma vitamin A, vitamin E, copper or zinc levels between animals with different types of foot lesions or with different severity scores. Shortly after the change to diet 2 (fed to juvenile captive flamingos that did not show any foot lesion), the flooring of the outdoor water pools was covered with fine granular sand. Because both factors (nutrition and flooring) were changed during the same evaluation period, it cannot be concluded which factor contributed in what extent to the reduction of foot lesions. While it is assumed that low plasma zinc levels identified in the group of juvenile captive flamingos on diet 1 were not directly responsible for foot lesions observed in these animals, they may have played a role in altering the skin integrity of the feet and predisposing them to pododermatitis.     

Identification of Chlamydia gallinacea in a parrot and in free‐range chickens in Australia

Chlamydia gallinacea is a recently described bacterial species in a genus known to infect and cause disease in animals and humans. Our report describes the identification of C. gallinacea infection in free‐range laying chickens (Gallus gallus) in Australia, and the identification of C. gallinacea infection in a parrot, a wild Australian galah (Eolophus roseicapillus). There is currently little knowledge of the effects of C. gallinacea infection on avian hosts, but it has been linked to respiratory disease in humans and could potentially cause similar disease in other species. Our report highlights the need for further study and surveillance of Chlamydia species in both wild and domestic hosts in Australia.     

Changes in in vitro ruminal and post‐ruminal degradation of tropical tannin‐rich legumes due to varying levels of polyethylene glycol

We evaluated the effects of tannins from Flemingia macrophylla (CIAT 17403) and Calliandra calothyrsus (San Ramón CIAT 22310 and Patulul CIAT 22316) on in vitro ruminal and post‐ruminal dry matter and apparent protein degradation. For each tannin source (legumes), different dosages of polyethylene glycol (PEG) (8000 Da) in McDougall buffer were added to achieve ratios of 0:3, 1:3, 2:3 and 3:3 PEG:condensed tannin (CT). Ruminal fluid mixed with McDougall buffer (1:4) was added to tubes containing only legume foliage (control) or PEG‐treated legume foliage. For both Calliandra varieties, a higher ruminal dry matter degradation was observed at a PEG:CT ratio of 3:3. For F. macrophylla, no differences were found between 2:3 and 3:3 ratios (p > 0.05), indicating that a PEG:CT ratio of 2:3 might be enough to bind tannins. Increasing PEG:CT ratios increased apparent ruminal degraded protein and ammonia concentration (p < 0.0001) differing among species (species × ratio: p < 0.0001). The degradation of bypass crude protein (dBCP) was influenced by both legume type and PEG:CT ratio (p < 0.0001). For Patulul, as PEG:CT ratio increased, dBCP increased, but after tannin ratio of 2:3, there was not a significant increase, and for San Ramón, dBCP degradation was higher as PEG:CT ratio increased up to 2:3. For Flemingia, dBCP was higher than PEG:CT ratio of 0:3 but not different among 1:3, 2:3 or 3:3. Low concentration of CT (116 mg/g DM) increased the proportion of protein digested in the abomasum, but higher levels of CT (252 mg/g) clearly reduced the proportion of digested CP. For Flemingia, PEG:CT ratio of 2:3 is enough to inactivate tannins, while PEG:CT ratio of 3:3 was needed for Calliandra and consequently increased ruminal degradation of dry mater (rdDM), and crude protein (rdCP), total degradation of dry matter (tdDM), crude protein (tdCP) and ammonia levels.     

Preliminary studies of alfaxalone for intravenous immobilization of juvenile captive estuarine crocodiles (Crocodylus porosus) and Australian freshwater crocodiles (Crocodylus johnstoni) at optimal and selected sub‐optimal thermal zones

ObjectivesTo investigate the character of immobilization given by alfaxalone in juvenile crocodiles at optimal and at suboptimal temperatures.Study designProspective, randomized partial crossover study.AnimalsTwenty captive male estuarine (weight 0.6–2.5 kg) and five captive male freshwater crocodiles (weight 0.2–0.6 kg).MethodsCrocodiles were acclimatized for 24 hours at one of the following environmental temperatures; 32 °C, 27 °C, 22 °C or 17 °C, then received 3 mg kg^?1 intravenous (IV) alfaxalone into the dorsal occipital venous sinus. Duration and quality of immobilization was assessed and heart rate (HR) measured. On a separate occasion each crocodile was immobilized at one other environmental temperature.ResultsAlfaxalone, 3 mg kg^?1 IV, produced immobilization for 55 (range 15–100 minutes in estuarine, and 20 (range 20–25) minutes in freshwater crocodiles at 32 °C. There was no significant difference overall in immobilization times between temperatures, other than that, in estuarine crocodiles, duration was shorter at 32 °C than 22 °C. The character of immobilization was unpredictable, with animals recovering without warning, or having extended recoveries requiring assisted ventilation. Assisted ventilation was necessary mainly at the lower temperatures. Median HR in all temperature treatments decreased within 5 minutes post–injection, but the change in HR over the duration of immobilization was affected by the temperature, with a progressively smaller range of fall as temperature decreased. At 17 °C, two estuarine crocodiles appeared to re–immobilize after initial recovery, became severely bradycardiac and required ventilation and re–warming.Conclusions and clinical relevanceAlfaxalone IV in small captive estuarine and freshwater crocodiles provides adequate induction of immobilization at various temperatures. However, the unpredictable results following induction mean it is unsuitable for field use and should be restricted to environments where intubation and ventilation are available, where animals can be warmed to optimal temperature, and where access to immersion in water can be restricted for 24 hours.     

Changes in the prevalence,genotypes and antimicrobial resistance phenotypes of non‐typhoidal Salmonella recovered from mail‐order hatchling poultry sold at US feed stores, 2013–2015

Every year, multiple outbreaks of salmonellosis in humans are linked to contact with mail‐order chicks and ducks. The objective of this study was to describe the temporal changes in the prevalence of serovars, genotypes and antimicrobial resistance (AMR) phenotypes of non‐typhoidal Salmonella (NTS) recovered from shipped boxes of mail‐order hatchling poultry in the United States during 2013 to 2015. In each year, a sample of feed stores belonging to a single national chain participated in the study. The store employees submitted swabs or hatchling pads from hatchling boxes and shipment tracking information of the arriving boxes to the investigators. NTS was cultured from the samples and isolates were sent to the National Veterinary Services Laboratories (Ames, IA) for serotyping, pulsed‐field gel electrophoresis (PFGE) and AMR phenotyping. The PFGE patterns of Salmonella serovars isolated from hatchling boxes were compared with those from human outbreaks of salmonellosis linked to live poultry contact. The box‐level prevalence of NTS was significantly higher in 2015 compared to 2014. Also, the population of Salmonella serovars recovered in 2015 was more diverse and substantially different from those recovered in the previous two years. Of PFGE patterns recovered from hatchling boxes, seven distinct patterns in 2015, three in 2014 and four in 2013 were indistinguishable from the PFGE patterns of human outbreaks‐associated strains in the respective years. Importantly, a significant positive correlation was found between the box‐level prevalence of PFGE patterns and the number of human illnesses associated with the same patterns. Also, the proportion of multidrug‐resistant isolates was higher in 2014 and 2015 compared to that in 2013. The results demonstrate that shipments of mail‐order hatchling poultry are frequently contaminated with Salmonella genotypes indistinguishable from human outbreaks‐associated strains each year, and control efforts at hatchery level are likely to have an important public health impact.     

A Cross‐Sectional Study Examining Campylobacter and Other Zoonotic Enteric Pathogens in Dogs that Frequent Dog Parks in Three Cities in South‐Western Ontario and Risk Factors for Shedding of Campylobacter spp.

An estimated 6 million pet dogs live in Canadian households with the potential to transmit zoonotic pathogens to humans. Dogs have been identified as carriers of Salmonella, Giardia and Campylobacter spp., particularly Campylobacter upsaliensis, but little is known about the prevalence and risk factors for these pathogens in pet dogs that visit dog parks. This study examined the prevalence of these organisms in the faeces of dogs visiting dog parks in three cities in south‐western Ontario, as well as risk factors for shedding Campylobacter spp. and C. upsaliensis. From May to August 2009, canine faecal samples were collected at ten dog parks in the cities of Guelph and Kitchener‐Waterloo, Ontario, Canada. Owners were asked to complete a questionnaire related to pet characteristics and management factors including age, diet and activities in which the dog participates. Faecal samples were collected from 251 dogs, and 189 questionnaires were completed. Salmonella, Giardia and Campylobacter spp. were present in 1.2%, 6.4% and 43.0% of faecal samples, respectively. Of the Campylobacter spp. detected, 86.1% were C. upsaliensis, 13% were C. jejuni and 0.9% were C. coli. Statistically significant sparing factors associated with the shedding of Campylobacter spp. included the feeding of a commercial dry diet and the dog's exposure to compost. Age of dog had a quadratic effect, with young dogs and senior dogs having an increased probability of shedding Campylobacter spp. compared with adult dogs. The only statistically significant risk factor for shedding C. upsaliensis was outdoor water access including lakes and ditches, while dogs >1 year old were at a lower risk than young dogs. Understanding the pet‐related risk factors for Campylobacter spp. and C. upsaliensis shedding in dogs may help in the development of awareness and management strategies to potentially reduce the risk of transmitting this pathogen from dogs to humans.     

Time‐coordinated prevalence of extracellular HGF,FGF2 and TGF‐β3 in crush‐injured skeletal muscle

Successful regeneration and remodeling of neuromuscular junctions are critical for restoring functional capacities and properties of skeletal muscle after damage, and axon‐guidance molecules may be involved in the signaling that regulates such restoration. Recently, we found that early‐differentiated satellite cells up‐regulate a secreted neural chemorepellent Sema3A upon in vivo muscle‐crush injury. The study also revealed that Sema3A expression is up‐regulated in primary satellite‐cell cultures in response to hepatocyte growth factor (HGF) and basic fibroblast growth factor (FGF2) and is prevented by transforming growth factor (TGF)‐β2, 3. In order to verify the physiological significance of this regulation in vitro, the present study was designed to estimate the time‐course of extracellular HGF, FGF2 and TGF‐β3 concentrations after crush‐injury of Gastrocnemius muscle in the rat lower hind‐limb, using a combination of a non‐homogenization/non‐spin extraction of extracellular wound fluids and enhanced chemiluminescence–Western blotting analyses. Results clearly demonstrated that active HGF and FGF2 are prevalent in 2–8 days post‐crush, whereas active TGF‐β3 increases after 12 days, providing a better understanding of the time‐coordinated levels of HGF, FGF2 and TGF‐β3 that drive regulation of Sema3A expression during regenerative intramuscular moto‐neuritogenesis.     

Serial Evaluation of Abdominal Fluid and Serum Amino‐terminal pro‐C‐type Natriuretic Peptide in Dogs with Septic Peritonitis

Background

Serum N‐terminal pro‐C‐natriuretic peptide (NT‐proCNP) has shown promise as a diagnostic biomarker for sepsis. Its sensitivity to detect dogs with septic peritonitis (SP) is reportedly low, perhaps attributable to the compartmentalization of NT‐proCNP in the abdominal cavity.

Objectives

To evaluate the use of an ELISA for the measurement of NT‐proCNP in canine abdominal fluid and to describe the peri‐operative pattern of abdominal fluid and serum NT‐proCNP concentrations in dogs with SP.

Animals

Five client‐owned dogs with nonseptic abdominal effusion of varying etiologies and 12 client‐owned dogs with SP undergoing abdominal surgery and placement of a closed‐suction abdominal drain (CSAD). Six dogs were included upon hospital admission; 6 were included the day after surgery.

Methods

Prospective pilot study. A commercially available ELISA kit was analytically validated for use on canine abdominal fluid. The NT‐proCNP concentrations were measured in the abdominal fluid of control dogs, and in serum and abdominal fluid of dogs with SP from admission for CSAD removal.

Results

In dogs with SP, admission abdominal fluid NT‐proCNP concentrations were lower than the concurrent serum concentrations (P = 0.031), and lower than control canine abdominal fluid concentrations (P = 0.015). Postoperatively, abdominal fluid NT‐proCNP concentrations remained lower than serum concentrations (P < 0.050), except on day 4.

Conclusions and Clinical Importance

The ELISA kit was able to measure NT‐proCNP in canine abdominal fluid. In dogs with SP, low serum NT‐proCNP concentrations cannot be explained by abdominal compartmentalization.