Phenotypic and genotypic characterization of antimicrobial resistance in faecal enterococci from wild boars (Sus scrofa)

The objective was to study the prevalence of antimicrobial resistance and the mechanisms implicated in faecal enterococci of wild boars in Portugal. One hundred and thirty-four enterococci (67 E. faecium, 54 E. hirae, 2 E. faecalis, 2 E. durans and 9 Enterococcus spp.) were recovered from 67 wild boars (two isolates/sample), and were further analysed. High percentages of resistance were detected for erythromycin, tetracycline, and ciprofloxacin (48.5%, 44.8%, and 17.9%, respectively), and lower values were observed for high-level-kanamycin, -streptomycin, chloramphenicol, and ampicillin resistance (9%, 6.7%, 4.5%, and 3.7%, respectively). No isolates showed vancomycin or high-level-gentamicin resistance. The erm(B), tet(M), aph(3')-IIIa, and ant(6)-I genes were demonstrated in all erythromycin-, tetracycline-, kanamycin-, and streptomycin-resistant isolates, respectively. Specific genes of Tn916/Tn1545 and Tn5397 transposons were detected in 78% and 47% of our tet(M)-positive enterococci, respectively. The tet(S) and tet(K) genes were detected in one isolate of E. faecium and E. hirae, respectively. Three E. faecium isolates showed quinupristin-dalfopristin resistance and the vat(E) gene was found in all of them showing the erm(B)-vat(E) linkage. Four E. faecium isolates showed ampicillin-resistance and all of them presented seven amino acid substitutions in PBP5 protein (461Q-->K, 470H-->Q, 485M-->A, 496N-->K, 499A-->T, 525E-->D, and 629E-->V), in relation with the reference one; a serine insertion at 466' position was found in three of the isolates. Faecal enterococci from wild boars harbour a variety of antimicrobial resistance mechanisms and could be a reservoir of antimicrobial resistance genes and resistant bacteria that could eventually be transmitted to other animals or even to humans.     

Eastern equine encephalitis in 9 South American camelids

BACKGROUND: Eastern equine encephalitis (EEE) virus is a mosquito-borne togavirus (alphavirus) that causes severe (often fatal) encephalitis in many mammalian species, but it has not been reported previously in South American camelids. Hypothesis: South American camelids can become naturally infected with EEE virus and show encephalitic signs similar to those observed in other affected species. ANIMALS: Nine cases (8 alpacas and 1 llama, aged 3.5 weeks to 12 years) were identified; 4 of 9 were 510 weeks old. All cases were from the East Coast of the United States and presented in late summer and fall. METHODS: A retrospective study was performed to include confirmed cases of EEE in camelids in North America before 2006. RESULTS: Eight of nine (89%) camelids died or were euthanized in extremis, with the mean time to death of 2 days. Clinical signs were consistent with encephalitis and included fever, lethargy, ataxia, seizures, recumbency, torticollis, opisthotonus, and vestibular signs. No consistent hematologic abnormalities were identified, and cerebrospinal fluid contained an increased protein concentration in the single camelid analyzed. No successful therapy was identified. EEE was confirmed by alphavirus detection by using immunohistochemistry (IHC) and polymerase chain reaction (PCR) in the central nervous system (CNS) and by serology. Findings included polioencephalitis with lymphocytic perivascular cuffing; neutrophil infiltration; gliosis; neuron satellitosis; necrosis; and edema, with intracytoplasmic alphavirus within neurons and glial cells. No virus was detected in extraneural tissues. CONCLUSIONS AND CLINICAL IMPORTANCE: In endemic areas, EEE should be considered a differential diagnosis for young and adult camelids with CNS disease. Brain histopathology with indirect IHC or PCR is diagnostic.     

Argyrophilic nucleolar organizing regions and Ki67 equally reflect proliferation in fine needle aspirates of normal, hyperplastic, inflamed, and neoplastic canine lymph nodes (n = 101)

BACKGROUND: The count of argyrophilic nucleolar organizing regions (AgNOR) has been considered a useful variable that reflects cellular proliferation in canine lymph nodes, but it has not been compared with other markers of proliferation. Hypothesis: Ki67 and AgNORs are equally useful as markers of tissue proliferation in fine needle aspirates of canine lymph nodes. ANIMALS: A total of 101 dogs. MATERIAL AND METHODS: Prospective, observational study of a convenience sample of dogs. Two smears were prepared for a May-Gruenwald-Giemsa stain and a Ki67/AgNOR double stain. In addition, CD3/CD79a immunostaining was performed when cytologic examination revealed a lymphoma. The dogs were grouped as normal (n = 26), reactive hyperplasia (n = 25), lymphadenitis (n = 31), and lymphoma (n = 19), based on the physical examination and the cytologic findings. The AgNOR count/cell, AgNOR area/cell and the percentage of cells staining positive for Ki67 were evaluated in 100-167 cells (median, 113 cells) by using automatic image analysis. RESULTS: Mean (SD) AgNOR counts/cell were 1.36 +/- 0.19 in normal dogs, 1.55 +/- 0.26 in lymphadenitis, 1.65 +/- 0.32 in reactive hyperplasia, and 3.67 +/- 1.08 in lymphoma. The percentage of Ki67 positive cells was 2.67 +/- 0.99% in normal lymph nodes, 5.04 +/- 3.34% in lymphadenitis, 5.36 +/- 2.14% in reactive hyperplasia, and 30.2 +/- 10.8% in lymphoma. All variables were significantly higher in dogs with lymphoma compared with the other groups (P < .0001). The sensitivity and the specificity of the AgNOR count for diagnosing lymphoma were 95 and 96% at a cutoff value of >2.04 AgNORs/cell. The cutoff value for the Ki67 positive cells was >10.40% (sensitivity, 95%; specificity, 98%). CONCLUSION AND CLINICAL IMPORTANCE: The results indicated that both AgNOR and Ki67 counts were good diagnostic tools for assessment of proliferation in aspirates of canine lymph nodes.     

Impact of seed blend and structured maize refuge on Helicoverpa zea (Lepidoptera: Noctuidae) potential phenological resistance development parameters in pupae and adults

BACKGROUND

Helicoverpa zea, an economic pest in the south-eastern United States, has evolved practical resistance to Bacillus thuringiensis (Bt) Cry toxins in maize and cotton. Insect resistance management (IRM) programs have historically required planting of structured non-Bt maize, but because of its low adoption, the use of seed blends has been considered. To generate knowledge on target pest biology and ecology to help improve IRM strategies, nine field trials were conducted in 2019 and 2020 in Florida, Georgia, North Carolina, and South Carolina to evaluate the impact of Bt (Cry1Ab + Cry1F or Cry1Ab + Cry1F + Vip3A) and non-Bt maize plants in blended and structured refuge treatments on H. zea pupal survival, weight, soil pupation depth, adult flight parameters, and adult time to eclosion.

RESULTS

From a very large sample size and geography, we found a significant difference in pupal mortality and weight among treatments in seed blends with Vip3A, implying that cross-pollination occurred between Bt and non-Bt maize ears. There was no treatment effect for pupation depth, adult flight distance, and eclosion time.

CONCLUSION

Results of this study demonstrate the potential impact of different refuge strategies on phenological development and survival of an important pest species of regulatory concern. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Effects of epidural administration of dexmedetomidine on the minimum alveolar concentration of isoflurane in dogs

Objective-To evaluate the effects of epidural administration of 3 doses of dexmedetomidine on isoflurane minimum alveolar concentration (MAC) and characterize changes in bispectral index (BIS) induced by nociceptive stimulation used for MAC determination in dogs. Animals-6 adult dogs. Procedures-Isoflurane-anesthetized dogs received physiologic saline (0.9% NaCl) solution (control treatment) or dexmedetomidine (1.5 [DEX1.5], 3.0 [DEX3], or 6.0 [DEX6] mug/kg) epidurally in a crossover study. Isoflurane MAC (determined by use of electrical nociceptive stimulation of the hind limb) was targeted to be accomplished at 2 and 4.5 hours. Changes in BIS attributable to nociceptive stimulation and cardiopulmonary data were recorded at each MAC determination. Results-With the control treatment, mean +/- SD MAC values did not change over time (1.57 +/- 0.23% and 1.55 +/- 0.25% at 2 and 4.5 hours, respectively). Compared with the control treatment, MAC was significantly lower at 2 hours (13% reduction) but not at 4.5 hours (7% reduction) in DEX1.5-treated dogs and significantly lower at 2 hours (29% reduction) and 4.5 hours (13% reduction) in DEX3-treated dogs. The DEX6 treatment yielded the greatest MAC reduction (31% and 22% at 2 and 4.5 hours, respectively). During all treatments, noxious stimulation increased BIS; but changes in BIS were correlated with increases in electromyographic activity. Conclusions and Clinical Relevance-In dogs, epidural administration of dexmedetomidine resulted in dose-dependent decreases in isoflurane MAC and that effect decreased over time. Changes in BIS during MAC determinations may not represent increased awareness because of the possible interference of electromyographic activity.     

Clinical and pharmacokinetic evaluation of S-ketamine for intravenous general anaesthesia in horses undergoing field castration

Background

Intravenous anaesthetic drugs are the primary means for producing general anaesthesia in equine practice. The ideal drug for intravenous anaesthesia has high reliability and pharmacokinetic properties indicating short elimination and lack of accumulation when administered for prolonged periods. Induction of general anaesthesia with racemic ketamine preceded by profound sedation has already an established place in the equine field anaesthesia. Due to potential advantages over racemic ketamine, S-ketamine has been employed in horses to induce general anaesthesia, but its optimal dose remains under investigation. The objective of this study was to evaluate whether 2.5 mg/kg S-ketamine could be used as a single intravenous bolus to provide short-term surgical anaesthesia in colts undergoing surgical castration, and to report its pharmacokinetic profile.

Results

After premedication with romifidine and L-methadone, the combination of S-ketamine and diazepam allowed reaching surgical anaesthesia in the 28 colts. Induction of anaesthesia as well as recovery were good to excellent in the majority (n = 22 and 24, respectively) of the colts. Seven horses required additional administration of S-ketamine to prolong the duration of surgical anaesthesia. Redosing did not compromise recovery quality. Plasma concentration of S-ketamine decreased rapidly after administration, following a two-compartmental model, leading to the hypothesis of a consistent unchanged elimination of the parent compound into the urine beside its conversion to S-norketamine. The observed plasma concentrations of S-ketamine at the time of first movement were various and did not support the definition of a clear cut-off value to predict the termination of the drug effect.

Conclusions

The administration of 2.5 mg/kg IV S-ketamine after adequate premedication provided good quality of induction and recovery and a duration of action similar to what has been reported for racemic ketamine at the dose of 2.2 mg/kg. Until further investigations will be provided, close monitoring to adapt drug delivery is mandatory, particularly once the first 10 minutes after injection are elapsed. Taking into account rapid elimination of S-ketamine, significant inter-individual variability and rapid loss of effect over a narrow range of concentrations a sudden return of consciousness has to be foreseen.     

Results of an online questionnaire to survey calf management practices on dairy cattle breeding farms in Austria and to estimate differences in disease incidences depending on farm structure and management practices

Background

Calf disease may result in great economic losses. To implement prevention strategies it is important to gain information on management and to point out risk factors. The objective of this internet based survey was to describe calf management practices on registered dairy breeding farms in Austria and to estimate differences in calf disease incidences depending on farm structure and management practices.

Results

A total of 1287 questionnaires were finally analysed (response rate 12.2 %). Herd characteristics and regional distribution of farms indicated that this survey gives a good overview on calf management practices on registered dairy farms in Austria. The median number of cows per farm was 20 (interquartile range 13–30). Significant differences regarding farm characteristics and calf management between small and large farms (≤20 vs >20 cows) were present. Only 2.8 % of farmers tested first colostrum quality by use of a hydrometer. Storing frozen colostrum was more prevalent on large farms (80.8 vs 64.2 %). On 85.1 % of the farms, whole milk, including waste milk, was fed to the calves. Milk replacer and waste milk were more often used on large farms. In accordance with similar studies from other countries, calf diarrhoea was indicated as the most prevalent disease. Multivariable logistic regression analysis revealed that herd size was associated with calf diarrhoea and calf respiratory tract disease, with higher risk of disease on large farms. Furthermore, feeding waste milk to the calves was associated with increasing calf diarrhoea incidence on farm. In the final model with calf respiratory tract disease as outcome, respondents from organic farms reported less often a respiratory tract disease incidence of over 10 % compared with conventional farms [odds ratio (OR) 0.40, 95 % confidence interval (CI) 0.21–0.75] and farmers that housed calves individually or in groups after birth significantly reported more often to have an incidence of respiratory tract disease >10 % compared with farms where all calves were housed individually (OR 2.28, 95 % CI 1.16–4.48).

Conclusion

The results obtained in this study provide an overview on calf management on dairy breeding farms in Austria and may help to further point out areas to be improved on farm.

Electronic supplementary material

The online version of this article (doi:10.1186/s13028-015-0134-y) contains supplementary material, which is available to authorized users.     

trans-Resveratrol, quercetin, (+)-catechin, and (-)-epicatechin content in south Italian monovarietal wines: relationship with maceration time and marc pressing during winemaking

The concentrations of trans-resveratrol, (+)-catechin, (-)-epicatechin, and quercetin were evaluated by means of high-performance liquid chromatography-diode array detection in red wines obtained from Aglianico, Piedirosso, and Nerello Mascalese grapes. The trans-resveratrol and epicatechin concentrations did not differ significantly between experimental wines. The concentration of quercetin in Nerello Mascalese wines was more than twice that observed in Aglianico and Piedirosso wines. Nerello Mascalese wines also significantly differed from other wines in the (+)-catechin content, which was significantly higher than those found in the other two wines. During maceration, the maximum extraction of trans-resveratrol was reached after 12 days for Aglianico and Piedirosso, after which a decline was observed. On the contrary, in the case of Nerello Mascalese, the concentration of trans-resveratrol increased steadily throughout the whole maceration process. After 2 days of maceration, the maximum concentration of quercetin was observed in Aglianico must, whereas the maximum quercetin extraction was reached after 12 days for Piedirosso and 17 days for Nerello Mascalese. The maximum levels of (+)-catechin and (-)-epicatechin were generally observed after 12 days of maceration for all wines, although a decline of (-)-epicatechin occurred after maximum extraction in Aglianico and Piedirosso wines. Following marc pressing, a significant increase in the concentration of trans-resveratrol for Aglianico, (+)-catechin and (-)-epicatechin for Piedirosso, and (-)-epicatechin for Nerello Mascalese was observed.