Antimicrobial Resistance and Current Alternatives in Veterinary Practice: A Review.

Antibiotics are commonly used to treat bacterial infections. For many years, antibiotics have been used at sub-therapeutic doses to promote animal growth and misused as prophylactics and metaphylactic on farms. The widespread and improper use of antibiotics has resulted in a serious problem, defined as antibiotic resistance by the World Health Organisation, which is a major public health threat in the 21st century. Bacteria have evolved sophisticated mechanistic strategies to avoid being killed by antibiotics. These strategies can be classified as intrinsic resistance (referring to the inherent structural or functional characteristics of a bacterial species) or acquired resistance (referring to mutations in chromosomal genes or the acquisition of external genetic determinants of resistance). In farm animals, the use of antibiotics warrants serious consideration, as their residues leach into the environment through effluents and come into contact with humans through food. Several factors have contributed to the emergence of antibiotic-resistant bacteria. This review provides an update on antibiotic resistance mechanisms, while focusing on the effects of this threat on veterinary medicine, and highlighting causal factors in clinical practice. Finally, it makes an excursus on alternative therapies, such as the use of bacteriophages, bacteriocins, antimicrobial photodynamic therapy, phytochemicals, and ozone therapy, which should be used to combat antibiotic-resistant infections. Some of these therapies, such as ozone therapy, are aimed at preventing the persistence of antibiotics in animal tissues and their contact with the final consumer of food of animal origin.

In-vitro antibiotic resistance phenotypes of respiratory and enteric bacterial isolates from weaned dairy heifers in California.

Antimicrobial drug (AMD) use for bovine respiratory disease (BRD) continues to be concerning for development of antimicrobial resistance (AMR) in respiratory and enteric bacteria of cattle. This study aimed to provide data regarding AMR in respiratory isolates, and identify relationships between respiratory and enteric AMD susceptibility, in weaned dairy heifers. A cross-sectional study was performed between June of 2019 and February 2020, on 6 calf rearing facilities in California. Deep nasopharyngeal and rectal swabs were collected from 341 weaned heifers and submitted for selective bacterial culture and AMR testing. Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni were selectively isolated from respiratory samples; Escherichia coli and Enterococcus spp. were selectively isolated from rectal swabs. Minimum inhibitory concentrations (MIC) were determined for selected isolates against 19 AMD. The proportion of resistant isolates was calculated using Clinical Laboratory Standards Institute (respiratory) or USDA NARMS (enteric) breakpoints; when no applicable breakpoint was available, the distribution of MIC was described and compared. Association between AMR in a calf's respiratory isolate and a higher or lower MIC of the matched enteric isolates was determined. More than 50% of P. multocida isolates were resistant to each of 7 AMD commonly used to treat BRD (florfenicol, gamithromycin, tildipirosin, tilmicosin, danofloxacin, enrofloxacin and tetracycline). Resistance in respiratory isolates was only associated with higher matched enteric MIC for gamithromycin and tulathromycin. Multidrug resistance was reported in >70% of P. multocida and M. haemolytica isolates. Antimicrobial resistance, including multidrug resistance, in respiratory isolates appears to be widespread in weaned dairy heifers; this finding has not previously been reported and raises concern for the future efficacy of AMD used to treat respiratory diseases in weaned dairy heifers. Enteric bacterial MIC appear to have limited direct association with respiratory isolate AMR classification.

Mitigating multiple stresses in Pangasianodon hypophthalmus with a novel dietary mixture of selenium nanoparticles and Omega-3-fatty acid.

Effects of a novel dietary mixture of selenium nanoparticles (Se-NPs) and omega-3-fatty acids i.e., Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on mitigating arsenic pollution, high-temperature stress and bacterial infection were investigated in Pangasianodon hypophthalmus. To aim this, four isocaloric and iso-nitrogenous diets were prepared: control feed (no supplementation), Se-NPs at 0.2 mg kg-1 diet with EPA + DHA at 0.2, 0.4 and 0.6% as supplemented diets. Fish were reared under normal condition or concurrent exposure to arsenic (2.65 mg L-1), and temperature (34 °C) (As + T) stress for 105 days. The experiment was conducted with eight treatments in triplicates. Response to various stresses i.e., primary (cortisol), secondary (oxidative stress, immunity, and stress biomarkers) and tertiary stress response (growth performance, bioaccumulation and mortality due to bacterial infection) were determined. Supplementation of dietary Se-NPs at 0.2 mg kg-1 diet and EPA + DHA at 0.2 and 0.4% reduced the primary stress level. Exposure to arsenic and temperature (As + T) and fed with control diet and EPA + DHA at 0.6% aggravated the cortisol level. Anti-oxidative enzymes (Catalase, superoxide dismutase, glutathione peroxidase and glutathione-s-transferase) and immunity (Nitroblue tetrazolium, total protein, albumin, globulin, A:G ratio, total immunoglobulin and myeloperoxidase) of the fish were augmented by supplementation of Se-NPs and EPA + DHA at 0.2 and 0.4%. Neurotransmitter enzyme, HSP 70, Vitamin C were significantly enhanced (p < 0.01) with supplementation of Se-NPs at 0.2 mg kg-1 and EPA + DHA at 0.2 and 0.4%. Whereas total lipid, cholesterol, phospholipid, triglyceride and very low-density lipoprotein (VLDL) were reduced (p < 0.01) with the supplementation of Se-NPs at 0.2 mg kg-1 diet and EPA + DHA at 0.2 and 0.4%. Tertiary stress response viz. growth performance was also significantly enhanced with supplementation of Se-NPs at 0.2 mg kg-1 and EPA + DHA at 0.2 and 0.4% reared under As + T. Whereas arsenic bioaccumulation in fish tissues was significantly reduced with dietary supplementation of Se-NPs and EPA + DHA. Cumulative mortality and relative percentage survival were reduced with Se-NPs at 0.2 mg kg-1 and EPA + DHA at 0.2 and 0.4%. The investigation revealed that a novel combination of Se-NPs at 0.2 mg kg-1 and EPA + DHA at 0.4% followed by 0.2% has the potential to alleviate temperature stress, bacterial infection and arsenic pollution. Whereas diet containing Se-NPs at 0.2 mg kg-1 diet and EPA + DHA at 0.6% was noticeably enhanced the stress in P. hypophthalmus.

Successful treatment of recurrent subclinical mastitis in cows caused by enrofloxacin resistant bacteria by means of the sequential intramammary infusion of enrofloxacin HCl-2H2O and ceftiofur HCl: a clinical trial.

BACKGROUND: Recurrent subclinical mastitis (RScM) due to resistant bacteria has low clinical and bacteriological cure rates, often requiring the culling of cows. The sequential intra-mammary administration of enrofloxacin hydrochloride-dihydrate (enro-C) followed by ceftiofur HCl may be useful for treating these cases. OBJECTIVES: This study assessed the bacteriological and clinical cure-efficacies of the sequentially intramammary administration of enro-C, followed by ceftiofur HCl to treat RScM in Holstein/Friesian cows. METHODS: This trial was conducted in a herd with a high prevalence of RScM, and 20 Holstein/Friesian cows were included: 45% suffering subclinical mastitis and 38.9% of the mammary quarters affected. Twenty-nine bacterial isolates in vitro resistant to enro-C were obtained (coagulase-negative Staphylococcus spp, 55.2%; Staphylococcus aureus, 27.6%; Escherichia coli, 6.9%; Streptococcus uberis, 6.9%; Corynebacterium bovis, 3.4%). Polymerase chain reaction-isolated the following genes linked to enro-C resistance: chromosomal (gyrA) and plasmid (aac(6')-lb-cr). The treatments were as follows: twice-daily intramammary infusions of enro-C (300 mg/10 mL) for 5 days. Cows clinically considered treatment failures were also treated with intramammary ceftiofur (125 mg/10 mL, twice daily for 5 days. The clinical and bacteriological cure rates were carried out when completing each treatment phase and at 14 and 21 days, aided by a California mastitis test, somatic cell count, and failure to identify the initially causative bacteria. RESULTS: Enro-C achieved 65% clinical and bacteriological cure rates, and 100% cure rates were obtained after the rescue treatment with ceftiofur HCl. CONCLUSIONS: Outstanding clinical and bacteriological cure rates in cows affected by RScM were achieved with the consecutive intramammary infusions of enro-C, followed by ceftiofur HCl.

Evaluation of fully oxidised ß-carotene as a feed ingredient to reduce bacterial infection and somatic cell counts in pasture-fed cows with subclinical mastitis.

AIMS: To assess the effect of oral supplementation with fully oxidised ß-carotene (OxBC) on bacteriological cure, the incidence of clinical mastitis, and somatic cell counts (SCC) in pasture-fed cows with subclinical intramammary infection. METHODS: Cows from four dairy herds were enrolled in early lactation if they had quarter-level SCC >200,000 cells/mL and they had a recognised bacterial intramammary pathogen in one or more quarters. They were randomly assigned to be individually fed from Day 0, for a mean of 40 days, with 0.5 kg of a cereal-based supplementary feed that either contained 300 mg of OxBC (treatment; n = 129 quarters) or did not (control; n = 135 quarters). Quarter-milk samples were collected on Days 21 and 42 for microbiology and SCC assessment. Bacteriological cure was defined as having occurred when the bacteria present on Day 0 were not isolated from samples collected on Days 21 or 42. A new intramammary infection was defined as a bacterial species isolated either on Day 21 or 42 differing from that isolated on Day 0. Clinical mastitis was diagnosed and recorded by herdowners up to Day 42. RESULTS: The bacteriological cure rate was greater for quarters from cows in the treatment group (13.9 (95% CI = 4.1-23.7)%) than for cows in the control group (6.9 (95% CI = 4.8-9.1)%; p = 0.02). The percentage of quarters that developed a new intramammary infection at Day 21 or 42 was greater for cows in the treatment group (17.9 (95% CI = 6.7-29.1)%) than for cows in the control group (13.0 (95% CI = 4.3-21.8)%; p < 0.01). The prevalence of quarters that were infected on Day 42 was less in cows in the treatment group (79.9 (95% CI = 62.3-97.6)%) than the control group (88.2 (95% CI = 78.4-97.9)%; p = 0.009). The incidence of quarters diagnosed with clinical mastitis by Day 42 was lower in cows from the treatment group (1/129 (0.78 (95% CI = 0.02-4.24)%)) than in cows from the control group (6/135 (4.44 (95% CI = 1.65-9.42)%; p = 0.03)). Mean quarter-level SCC was not statistically different between treatment groups (p = 0.34). CONCLUSIONS AND CLINICAL RELEVANCE: Feeding 300 mg/cow/day of OxBC resulted in a higher bacteriological cure rate, a lower prevalence of intramammary infection, and a lower incidence of clinical mastitis compared to untreated controls. However new intramammary infections increased in treated cows, and the magnitude of the increased bacteriological cure was low, resulting in 80% of cows remaining infected at Day 42. Therefore treatment with OxBC should be considered as an adjunct to other mastitis control measures.

Calcium sulfate antibiotic-impregnated bead implantation for deep surgical site infection associated with orthopedic surgery in small animals.

OBJECTIVE: To report the outcomes and complications associated with antibiotic-impregnated calcium sulfate beads for prevention and treatment of orthopedic-related surgical site infection (SSI) in companion animals. STUDY DESIGN: Retrospective case series. ANIMALS: Client-owned cats (n = 2) and dogs (n = 14). METHODS: Medical records of 16 cases in which implantation of antibiotic-impregnated calcium sulfate beads was performed for the prevention or treatment of SSI were reviewed. Information collected included signalment, prior surgery, reason for bead placement, antibiotics used, bacterial culture results, and clinical outcomes. RESULTS: Surgical site infection resolved in six of 10 animals treated therapeutically and did not occur in six of six animals treated prophylactically. Susceptibility of the causative bacteria to the antibiotic implanted was confirmed in five of six cases with resolved SSI treated therapeutically but in only one of four cases with unresolved SSI treated therapeutically. Complications directly related to bead placement were evident in only one case in which beads extruded from external skeletal fixator pin tracts 7 days after implantation. At final follow-up, 11 of 12 animals without SSI had satisfactory limb use and no clinical, cytologic, or radiographic evidence of infection. CONCLUSION: Implantation was well tolerated. Resolution of SSI was inconsistent; however, when bacteria were susceptible to the antibiotic implanted, SSI resolved in all but one case. CLINICAL SIGNIFICANCE: Antibiotic-impregnated calcium sulfate beads could be considered for prevention or treatment of orthopedic SSI in small animals. A prospective clinical study is required to obtain additional information, including the value of preoperative bacterial culture.

Dietary Azomite, a natural trace mineral complex, improved the growth, immunity response, intestine health and resistance against bacterial infection in largemouth bass (Micropterus salmoides).

Azomite is a hydrated calcium sodium aluminosilicat rich in rare earth elements. To investigate the dietary effects of Azomite on growth, intestine microbiota and morphology, immunohematological changes and disease resistance, seven diets with Azomite supplementation of 0 (the control), 1.0, 2.0, 3.0, 4.0, 5.0 and 6.0 g/kg (A0, A1, A2, A3, A4, A5, A6), were prepared and fed to largemouth bass, Micropterus salmoides (7.96 ± 0.19) for 60 days. The results revealed that the weight gain (WG) increased first and then decreased with the increasing dietary Azomite, and the A2 group presented the highest WG and lowest feed conversion ratio among all the groups. The supplementation of 2.0 g/kg Azomite significantly increased the intestine protease activity, the crude protein of whole body and protein retention (P < 0.05), and high inclusion of Azomite (6.0 g/kg) significantly reduced the lipid retention (P < 0.05). The amounts of red blood cells in A5, A6 groups, white blood cells in A3, A5, A6 groups and lymphocyte in A2-A6 groups were all significantly higher than those in the control group (P < 0.05). In addition, serum superoxide dismutase and catalase activities in A5, A6 groups, and serum alkaline phosphatase and lysozyme activities in A2-A4 groups showed significantly higher values than the control group (P < 0.05). Intestinal microbiota analysis indicated that the Tenericutes abundance was increased, whereas Proteobacteria abundance was decreased in all Azomite supplemented groups. The villus height in A2-A4 groups, and the villus width in A2 group were significantly higher than those of the control group (P < 0.05). The cumulative mortality was reduced by the addition of 2.0-5.0 g/kg Azomite after challenging with A. hydrophila (P < 0.05). In conclusion, proper addition of Azomite in diets improved the growth, intestine morphology, immune response and disease resistance in largemouth bass, and the optimal inclusion was estimated to be 2.0-3.0 g/kg diet.

Bacterial Causes for Mortality Syndrome in Some Marine Fish Farms with Treatment Trials.

BACKGROUND AND OBJECTIVE: Bacterial fish diseases constitute a major problem in aquaculture, it was found in the environment and under stressors cause severe economic losses to fish. This work aimed to investigate the bacterial causes and suitable treatments of mass mortality in some cultured marine fish farms in Damietta governorate. MATERIALS AND METHODS: The study was performed on 5 farms suffered from mass mortality. Total of 100 diseased fish (10 sea bass and 10 sea bream/farm) and 20 water samples were randomly collected from these farms. Bacteriological examinations were carried out followed by in vitro sensitivity tests. Treatment trial was performed using the most effective antibacterial agent on isolated bacteria. RESULTS: From fish and water samples Pseudomonas spp., Aeromonas spp. and Vibrio spp. were isolated with the rat of (16, 10%), (22, 10%) and (28, 10%) respectively. These results were confirmed biochemically. Some virulence genes of isolated bacteria were detected using PCR; meanwhile, enrofloxacin reduced significantly the mortality rates in examined farms. CONCLUSION: It could be concluded that, Pseudomonas spp., Aeromonas spp. and Vibrio spp. are the main bacterial species causing mass mortality in marine fish farms. These bacteria were highly sensitive to enrofloxacin in vitro and in vivo.

A participatory surveillance of marsh deer (Blastocerus dichotomus) morbidity and mortality in Argentina: first results.

BACKGROUND: In an era of unprecedented socio-ecological changes, managing wildlife health demands high-quality data collection and the engagement of local communities. Blastocerus dichotomus, the largest South American deer, is Vulnerable to extinction mainly due to habitat loss. Diseases have been recognised as a potential threat, and winter mortality has been historically described in marsh deer populations from Argentina. Field difficulties have, however, prevented in-depth studies of their health status. RESULTS: Between May 2014 and April 2017, we investigated marsh deer morbidity and mortality in the two largest populations in Argentina. We collected data by means of a passive surveillance system that involved a network of researchers, field partners (veterinarians, park rangers, and local community), and decision makers. We sampled marsh deer during as well as outside mortality events. A total of 44 marsh deer with different body condition scores were evaluated. We obtained haematology and biochemistry values from animals with good body condition score. Marsh deer with poor body condition had a high burden of the ticks Amblyomma triste and Rhipicephalus microplus. Vector-borne agents such as Theileria cervi, Trypanosoma theileri, Trypanosoma evansi, Ehrlichia chaffeensis, Anaplasma platys, Anaplasma odocoilei, Anaplasma marginale, and Candidatus Anaplasma boleense were also found. Haemonchus spp., Ostertagia spp., and Trichostrongylus spp. were the most frequent gastrointestinal parasites in deer with poor body condition. A Multiple Correspondence Analysis reinforced a possible association of winter period with lower body score condition, high tick loads, infection with E. chaffeensis, and presence of harmful gastrointestinal parasites. CONCLUSIONS: Our approach allowed the establishment of a participatory surveillance network of marsh deer morbidity and mortality in Argentina. We report and analyse the first data obtained opportunistically within the framework of this network, providing information on the infectious and parasitic agents in marsh deer populations. The occurrence of Fasciola hepatica and Leptospira interrogans serovar pyrogenes is reported for the first time in wild marsh deer from Argentina. Our data will be useful to improve the interpretation of future mortality events. The field implementation of a surveillance network is key to a holistic approach to wildlife diseases.

Systematic Review and Meta-Analysis of the Occurrence of ESKAPE Bacteria Group in Dogs, and the Related Zoonotic Risk in Animal-Assisted Therapy, and in Animal-Assisted Activity in the Health Context.

Animal-assisted interventions are widely implemented in different contexts worldwide. Particularly, animal-assisted therapies and animal-assisted activities are often implemented in hospitals, rehabilitation centers, and other health facilities. These interventions bring several benefits to patients but can also expose them to the risk of infection with potentially zoonotic agents. The dog is the main animal species involved used in these interventions. Therefore, we aimed at collecting data regarding the occurrence of the pathogens ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter spp.) in dogs, in order to draft guidelines concerning the possible monitoring of dogs involved in animal-assisted therapies and animal-assisted activities in healthcare facilities. We performed a literature search using the PRISMA guidelines to examine three databases: PubMed, Web of Science, and Scopus. Out of 2604 records found, 52 papers were identified as eligible for inclusion in the review/meta-analysis. Sixteen papers reported data on E. faecium; 16 on S. aureus; nine on K. pneumoniae; four on A. baumannii; eight on P. aeruginosa; and six on Enterobacter spp. This work will contribute to increased awareness to the potential zoonotic risks posed by the involvement of dogs in animal-assisted therapies, and animal-assisted activities in healthcare facilities.

Reasons for antimicrobial treatment failures and predictive value of in-vitro susceptibility testing in veterinary practice: An overview.

The choice of the most suitable antimicrobial agent for the treatment of an animal suffering from a bacterial infection is a complex issue. The results of bacteriological diagnostics and the in-vitro antimicrobial susceptibility testing (AST) provide guidance of potentially suitable antimicrobials. However, harmonized AST methods, veterinary-specific interpretive criteria and quality control ranges, which are essential to conduct AST in-vitro and to evaluate the corresponding results lege artis, are not available for all antimicrobial compounds, bacterial pathogens, animal species and sites of infection of veterinary relevance. Moreover, the clinical benefit of an antimicrobial agent (defined as its in vivo efficacy) is not exclusively dependent on the in-vitro susceptibility of the target pathogen. Apart from the right choice of an antibacterial drug with suitable pharmacokinetic properties and an appropriate pharmaceutical formulation, the success of treatment depends substantially on its adequate use. Even if this is ensured and in-vitro susceptibility confirmed, an insufficient improvement of clinical signs might be caused by biofilm-forming bacteria, persisters, or specific physicochemical conditions at the site of infection, such as pH value, oxygen partial pressure and perfusion rate. This review summarizes relevant aspects that have an impact on the predictive value of in-vitro AST and points out factors, potentially leading to an ineffective outcome of antibacterial treatment in veterinary practice. Knowing the reasons of inadequate beneficial effects can help to understand possible discrepancies between in-vitro susceptibility and in vivo efficacy and aid in undertaking strategies for an avoidance of treatment failures.

L-lactate in cerebrospinal fluid can be used as a biomarker of encephalitis in cattle.

Cerebrospinal fluid (CSF) changes are significant for antemortem diagnoses of some neurological diseases. The aim of this study was to evaluate if the concentration of L-lactate in CSF could be used to differentiate healthy from encephalitic cattle. Cerebrospinal fluid samples from healthy cattle (n = 10) and from those naturally affected by rabies (n = 15), bovine herpesvirus type 5 meningoencephalitis (n = 16), histophilosis (n = 6), or bacterial encephalitis (n = 4), including 1 case of listeriosis, were collected and analyzed. Physical, biochemical (i.e., protein and glucose), and cellular analyses were performed in fresh samples. L-lactate, electrolytes (sodium, potassium, and chloride), calcium, and magnesium concentrations were measured in CSF samples that were kept frozen. L-lactate concentrations were also measured in plasma. Analysis of variance was used for comparison between groups and receiver operating characteristic analysis was performed considering L-lactate in CSF of healthy versus encephalitic cattle. The CSF L-lactate concentration was significantly higher in cattle with bacterial encephalitis than in healthy cattle; however, it did not differ between viral and bacterial encephalitis. The calcium concentrations were lower in cattle with encephalitis. L-lactate concentration in CSF > 3.6 mmol/L can be accepted as a cut-off value to indicate encephalitis. Thus, L-lactate in CSF is important for the diagnosis of encephalitis in cattle. Despite the small number of cases of bacterial encephalitis, it is suggested that L-lactate was not important for the differentiation between viral and bacterial encephalitis. Additional studies with a greater number of observations are necessary to clarify this, specifically in cases of listeriosis.

Les modifications du liquide céphalorachidien (LCR) sont importantes pour le diagnostic antemortem de certaines maladies neurologiques. Le but de cette étude était d'évaluer si la concentration de L-lactate dans le LCR pouvait être utilisée pour différencier les bovins en bonne santé des bovins encéphalitiques. Des échantillons de LCR provenant de bovins en bonne santé (n = 10) et de sujets infectés naturellement par la rage (n = 15), de méningoencéphalite à BoHV-5 (n = 16), l'histophilose (n = 6), ou d'encéphalite bactérienne (n = 4), notamment un cas de listériose ont été collectés et analysés. Des analyses physiques, biochimiques (protéines et glucose), et cellulaires ont été effectuées dans des échantillons frais. Les concentrations de L-lactate, d'électrolytes (Na+, K+, et Cl−), de calcium (Ca), et de magnésium ont été mesurées dans des échantillons de LCR maintenus congelés. Les concentrations de L-lactate ont également été mesurées dans le plasma. Une analyse de variance a été utilisée pour la comparaison entre les groupes et une analyse ROC (Receiver Operating Characteristic) a été réalisée en considérant le L-lactate dans le LCR de bovins en bonne santé par rapport à des bovins encéphalitiques. La concentration de L-lactate dans le LCR était significativement plus élevée chez les bovins présentant une encéphalite bactérienne que chez les bovins en bonne santé. Cependant, elle ne différait pas entre les bovins présentant une encéphalite virale et bactérienne. Les concentrations de Ca étaient plus faibles chez les bovins atteints d'encéphalite. Une concentration de L-lactate dans le LCR > 3,6 mmol/L peut être acceptée comme valeur seuil indiquant une encéphalite. Ainsi, le L-lactate dans le LCR est important pour le diagnostic de l'encéphalite chez les bovins. Malgré le petit nombre de cas d'encéphalite bactérienne inclus, il a été suggéré que la concentration de L-lactate dans le LCR dans la présente étude n'était pas une méthode de diagnostic important dans la différenciation entre l'encéphalite virale et bactérienne chez les bovins. Des études supplémentaires comportant un plus grand nombre d'observations sont nécessaires pour clarifier cet aspect, en particulier dans les cas de listériose.(Traduit par les auteurs).

Chemical composition and antibacterial activity of essential oils against pathogens often related to cattle endometritis.

INTRODUCTION: Endometritis is a condition marked by inflammation of the endometrium that affects dairy cows from 21 days after parturition, causing damage to herd fertility and economic losses on farms. The use of active compounds obtained from plant sources has gained importance as disease treatment agents in farm animals due to the high resistance rates currently observed against traditional antibiotics commonly used. The study was carried out to examine the chemical composition and to investigate the antibacterial activity of rosemary, cinnamon, cloves, eucalyptus, lemon, oregano and thyme essential oils against the reference strain of Escherichia coli (ATCC 25922), Fusobacterium necrophorum (ATCC 25286), Trueperella pyogenes (ATCC 19411) and Staphylococcus aureus (ATCC 29213), considered as typical bacteria causing endometritis. METHODOLOGY: The chemical composition of the seven essential oils were analyzed by GC-MS and their antibacterial activity was evaluated by the disc diffusion method. RESULTS: Thirty-six components were identified in total using GC-MS analyzes. The main compounds were cinnamaldehyde (86.5% for cinnamon essential oil), eugenol (85.7% for clove essential oil), 1,8-cineol (80% for eucalyptus and 47.8% rosemary essential oils), limonene (65.5% for lemon essential oil), carvacrol (72.1% for oregano essential oil) and thymol (48.8% for thyme essential oil). The disc diffusion assay revealed that cinnamon, clove, oregano, and thyme essential oils showed the best results compared to the other three essential oils, showing the largest zone of inhibition against all bacteria evaluated. CONCLUSIONS: These findings indicated that essential oils are a potential agent to be used as an alternative for bovine endometritis treatment.

Combination of herbal components (curcumin, carvacrol, thymol, cinnamaldehyde) in broiler chicken feed: Impacts on response parameters, performance, fatty acid profiles, meat quality and control of coccidia and bacteria.

The objective of this study was to determine whether curcumin and a commercial microencapsulated phytogenic supplement containing thymol, cinnamaldehyde and carvacrol in broiler chicken feed would improve health and meat quality (fatty acid profile), as well as to determine the coccidiostatic and bactericidal potential of the additives. The broiler chickens were divided into five groups: NC - negative control feed; PC - positive control; CU - with 50 mg/kg of curcumin, PHY - 100 mg/kg phytogenic; and PHY + CU, a combination of both additives at 50 mg/kg (curcumin) and 100 mg/kg (phytogenic). We observed significantly higher levels of total proteins associated with increased circulating globulins, as well as lower levels of uric acid, cholesterol and triglycerides in the PHY + CU group than in the NC. There were significantly fewer oocysts in birds supplemented with additives in the NC group on day 21; on day 35, the NC, PHY and PHY + CU groups had significantly lower counts than the PC and CU groups; however, at 44 days, the lowest counts were in PC group. The bacterial counts were significantly lower on day 21 in all groups that received additives than those of the control group; however, at 44 days, the bacterial and Escherichia coli counts in these groups were significantly higher than those of the control. Curcumin with or without phytogenic agent improved meat quality, with increased antioxidant levels and reduction of lipid peroxidation. There were significantly lower total saturated fatty acid levels and significantly greater monounsaturated/polyunsaturated fatty acid levels in broilers that consumed additives individually and in combination. The combination of additives significantly increased the crypt/villus ratio, a marker of improved intestinal health and performance. Additives potentiated their individual effects, suggesting they can replace conventional growth promoters without compromising health, intestinal mucosa or meat quality.

Antibiotic resistance in porcine pathogenic bacteria and relation to antibiotic usage.

BACKGROUND: Optimal treatment and prudent use of antimicrobials for pigs is imperative to secure animal health and prevent development of critical resistance. An important step in this one-health context is to monitor resistance patterns of important animal pathogens. The aim of this study was to investigate the antimicrobial resistance patterns of five major pathogens in Danish pigs during a period from 2004 to 2017 and elucidate any developments or associations between resistance and usage of antibiotics. RESULTS: The minimum inhibitory concentration (MIC) for Escherichia coli, Actinobacillus pleuropneumoniae, Streptococcus suis, Bordetella bronchiseptica, and Staphylococcus hyicus was determined to representatives of antibiotic classes relevant for treatment or surveillance. Escherichia coli isolates were mostly sensitive to fluoroquinolones and colistin, whereas high levels of resistance were observed to ampicillin, spectinomycin, streptomycin, sulfonamides and tetracycline. While resistance levels to most compounds remained relatively stable during the period, resistance to florfenicol increased from 2.1% in 2004 to 18.1% in 2017, likely in response to a concurrent increase in usage. A temporal association between resistance and usage was also observed for neomycin. E. coli serovars O138 and O149 were generally more resistant than O139. For A. pleuropneumoniae, the resistance pattern was homogenous and predictable throughout the study period, displaying high MIC values only to erythromycin whereas almost all isolates were susceptible to all other compounds. Most S. suis isolates were sensitive to penicillin whereas high resistance levels to erythromycin and tetracycline were recorded, and resistance to erythromycin and trimethoprim increasing over time. For S. hyicus, sensitivity to the majority of the antimicrobials tested was observed. However, penicillin resistance was recorded in 69.4-88.9% of the isolates. All B. bronchiseptica isolates were resistant to ampicillin, whereas all but two isolates were sensitive to florfenicol. The data obtained have served as background for a recent formulation of evidence-based treatment guidelines for pigs. CONCLUSIONS: Antibiotic resistance varied for some pathogens over time and in response to usage. Resistance to critically important compounds was low. The results emphasize the need for continuous surveillance of resistance patterns also in pig pathogenic bacteria.

Grape Pomace as a Promising Antimicrobial Alternative in Feed: A Critical Review.

Antimicrobial resistance is among the most urgent global challenges facing sustainable animal production systems. The use of antibiotics as growth promoters and for infectious disease prevention in intensive animal-farming practices has translated into the selection and spread of antimicrobial resistance genes in an unprecedented fashion. Several multi-resistant bacterial strains have been isolated from food-producing animals, thus constituting an alarming food-safety issue. Many industrial byproducts with potential antimicrobial properties are currently being investigated to identify empirical and affordable solutions/alternatives that can potentially be used in feed for animals. Grape pomace is among such byproducts that gained the attention as a result of its low cost, abundance, and, most importantly, its bioactive and antibacterial properties. This review discusses the recently reported studies with regard to exploring the use of grape pomace (and its extracts) in animal production to control pathogens, along with the promotion of beneficial bacterial species in the gut to ultimately alleviate antibacterial resistance. The review further summarizes realistic expectations connected with grape pomace usage and lists the still-to-be-addressed concerns about its application in animal agriculture.

Usage patterns of carbapenem antimicrobials in dogs and cats at a veterinary tertiary care hospital.

BACKGROUND: Carbapenems are a class of antimicrobials reserved for resistant infections or systemically ill people, yet the extent and context in which they are prescribed in the small animals is understudied. HYPOTHESIS/OBJECTIVE: To describe cases in dogs and cats treated with carbapenems to establish baseline data regarding the types of infections, outcomes, and resistance profiles of target infections. We hypothesize that prescribing practices for carbapenems at a veterinary tertiary care hospital would not comply with the recommended use guidelines in human medicine. METHODS: Retrospective study of veterinary medical records from all dogs and cats prescribed carbapenems between May 1, 2016, and April 30, 2017. RESULTS: A total of 81 infections (71 in dogs and 10 in cats) representing 68 animals (58 dogs and 10 cats) involving carbapenem use were identified. Cultures were performed in 65/81 (80%) infections, and antimicrobial use was de-escalated or discontinued in 10/81 (12%) infections. The average duration of treatment was 27.5 days and ranged from 1 to 196 days. Resistance to more than 3 antimicrobial classes was present in 57/115 (50%) isolates. Resistance to carbapenems was found in 2/64 (3%) of the bacterial isolates with reported carbapenem susceptibility. CONCLUSIONS AND CLINICAL IMPORTANCE: The majority of carbapenem use at a veterinary tertiary care hospital was prescribed in conjunction with culture and sensitivity determination, with de-escalation performed in a minority of cases, and treatment durations longer than typically recommended in human medicine.

Questions associated with the development of novel drugs intended for the treatment of bacterial infections in veterinary species.

The emergence of multi-drug resistant bacteria has limited therapeutic options for the treatment of bacterial diseases in both human and veterinary medicine. This has resulted in an urgent need for novel agents to treat infectious diseases. Veterinary medicine is further constrained by the need to ensure that our emerging therapeutics have minimal or no impact on resistance in human pathogens. Thus, there has recently been increased attention given to the development of alternative treatments for infectious disease in animals. The domain of alternative therapies, which includes antimicrobial peptides, bacteriophages, probiotics, and immunomodulators, provides a means to directly inhibit the ability of a pathogen to damage the host while optimally, not imposing a selective pressure favouring antibiotic resistance. However, it is recognized that bacterial pathogens have the capability of expressing a variety of virulence factors, necessitating a clear understanding of the specific target for that therapeutic intervention. This manuscript explores the various virulence mechanisms, the potential utility of developing novel anti-virulence agents for counteracting the expression of diseases associated with veterinary species, and some of the unique regulatory hurdles to be addressed within the framework of a new animal drug application. We conclude with the public health concerns to be considered as these agents are integrated into the veterinary therapeutic arsenal. Our hope is that this manuscript will provide a platform to stimulate discussions on the critical questions that need to be addressed.

Antibacterial activity against porcine respiratory bacterial pathogens and in vitro biocompatibility of essential oils.

Bacterial respiratory infections affecting pigs such as pneumonia, pleuropneumonia, and pleurisy, are a major health concern in the swine industry and are associated with important economic losses. This study aimed to investigate the antibacterial activities of essential oils against major swine respiratory pathogens with a view to developing a potential alternative to antibiotics. Their synergistic interactions with the bacteriocin nisin was also examined. Lastly, we assessed the in vitro biocompatibility of the most efficient essential oils using a pig tracheal epithelial cell line. Of the nine essential oils tested, those from cinnamon, thyme, and winter savory were the most active against Streptococcus suis, Actinobacillus pleuropneumoniae, Actinobacillus suis, Bordetella bronchiseptica, Haemophilus parasuis, and Pasteurella multocida, with minimum inhibitory concentrations and minimum bactericidal concentrations ranging from 0.01 to 0.156% (v/v). The main component found in cinnamon, thyme, and winter savory oils were cinnamaldehyde, thymol, and carvacrol, respectively. Treating pre-formed S. suis and A. pleuropneumoniae biofilms with thyme or winter savory oils significantly decreased biofilm viability. We also observed a synergistic growth inhibition of S. suis with mixtures of nisin and essential oils from thyme and winter savory. Concentrations of nisin and cinnamon, thyme and winter savory essential oils that were effective against bacterial pathogens had no effect on the viability of pig tracheal epithelial cells. The present study brought evidence that essential oils are potential antimicrobial agents against bacteria associated with porcine respiratory infections.

Mechanism of Brassica oleracea performance in bovine infectious mastitis by bioinformatic analysis.

Bovine mastitis affects dairy cattle worldwide and, despite the existing therapeutic measures, is not totally under control, leading to the need to develop alternative strategies. Brassica oleracea is a phytochemical commonly used in the control and prevention of human and animal diseases. The use of this plant in the treatment of infectious bovine mastitis has been little referenced in the literature and its molecular mechanism of action in this disease has not been clarified yet. This study aimed to reveal, through bioinformatic analysis, the molecular mechanism of action of Brassica oleracea in bovine mastitis. We investigated genes expressed in the signaling pathways of bovine mastitis and Brassica oleracea performance and elaborated the Venn diagram. A gene network was developed using the STRING 10 database. Leader genes were identified by calculating the weighted number of links (WNL). The NetworkAnalyzer plugin for Cytoscape software was used to characterize network topology. For the visualization of highly interconnected regions in the network, the MCODE was used. The BINGO and GFD-Net plugins were used to perform the ontological analysis. The TP53 and MTOR leader genes were identified in the sub-networks of the bovine mastitis signaling pathway and Brassica oleracea performance, respectively. Topological analysis confirmed the leader condition of the genes. Although the overlap of genes in the Venn diagram was not observed, the leader genes were found to be interconnected (confidence = 0.9). In the network that interconnected the leader genes two molecular complexes were detected and the ontological analysis revealed biological processes, cellular components and important molecular functions. It was concluded that Brassica oleracea may be a promising candidate to be included in a mammalian herbal cocktail against infectious bovine mastitis by interfering in the mechanisms of action of genes such as MTOR and TP53.