Antimicrobial susceptibility of bacteria isolated from neonatal foal samples submitted to a New Zealand veterinary pathology laboratory (2004 to 2013)

AIMS: To describe antimicrobial susceptibility, and identify antimicrobial resistance (AMR), in bacteria isolated from New Zealand foals.

METHODS: A database search was performed of submissions to a veterinary pathology laboratory between April 2004 and December 2013 for bacterial culture of samples from foals <3 weeks of age. Culture and susceptibility results were compiled with demographic information. Susceptibility results were as defined for the Kirby-Bauer disk diffusion susceptibility test based on Clinical Laboratory Standards Institute guidelines. Multi-drug resistance (MDR) was defined as non-susceptibility to ≥3 of a panel of antimicrobials (ceftiofur, enrofloxaxin, gentamicin, penicillin, tetracycline, trimethoprim-sulfonamide); penicillin susceptibility was not included for Gram-negative isolates.

RESULTS: Submissions from 102 foals were examined, and 127 bacterial isolates were cultured from 64 (63%) foals. Of the 127 isolates, 32 (25%) were Streptococcus spp., 30 (24%) were Staphylococcus spp., 12 (10%) were Enterococcus spp. and 26 (21%) were Escherichia coli. Of 83 Gram-positive isolates, 57 (69%) were susceptible to penicillin. Over all isolates, 92/126 (73%) were susceptible to gentamicin and 117/126 (93%) to enrofloxacin; 62/82 (76%) of Gram-positive, and 22/42 (52%) of Gram-negative bacteria were susceptible to ceftiofur; 53/81 (65%) of Gram-positive, and 23/44 (52%) of Gram-negative bacteria were susceptible to tetracycline; 59/82 (72%) of Gram-positive, and 23/44 (43%) of Gram-negative bacteria were susceptible to trimethoprim-sulfonamide. Of 126 isolates, 33 (26%) had MDR; >1 isolate with MDR was cultured from 24/64 (38%) foals, and ≥2 isolates with MDR were recovered from 8/64 (13%) foals.

CONCLUSIONS: Multi-drug resistance, including resistance to commonly used antimicrobials, was found in bacterial isolates from foals in New Zealand.

CLINICAL RELEVANCE: The results of this study are of concern from a treatment perspective as they indicate a potential for antimicrobial treatment failure. For future surveillance of AMR and the creation of national guidelines, it is important to record more data on samples submitted for bacterial culture.     

Breed influences on in vitro development of abattoir-derived bovine oocytes

Background

There is a discrepancy in the reproductive performance between different cattle breeds. Using abattoir-derived ovaries and data base information we studied the effects of breed on in vitro fertilization and early embryo development.

Methods

The in vitro developmental competence of oocytes from cattle (n = 202) of Swedish Red (SR), Swedish Holstein (SH) and mixed beef breeds was compared, retrospectively tracing donors of abattoir-derived ovaries using a combination of the national animal databases and abattoir information. Age was significantly lower and carcass conformation score was higher in the beef breeds than in the dairy breeds.Cumulus oocyte complexes (n = 1351) were aspirated from abattoir-derived ovaries from animals of known breed (visual inspection confirmed through databases), age (databases), and abattoir information. Oocytes were matured, fertilized (frozen semen from two dairy bulls) and cultured according to conventional protocols. On day 8, blastocysts were graded and the number of nuclei determined.

Results

Cleavage rate was not different between the breeds but was significantly different between bulls. The percentage of blastocysts on day 8 was significantly higher when the oocyte donor’s breed was beef or SR than SH. There was no significant difference in blastocyst grades or stages between the breeds, but the number of nuclei in day 8 blastocysts was significantly lower in SH compared to the beef.

Conclusions

The use of abattoir-derived ovaries from animals whose background is traceable can be a valuable tool for research. Using this approach in the present study, oocyte donor breed was seen to affect early embryo development during in vitro embryo production, which may be a contributing factor to the declining fertility in some dairy breeds seen today.     

Dysoxylum binectariferum Hook.f (Meliaceae), a rich source of rohitukine

Rohitukine, a chromane alkaloid, is a precursor of flavopiridol, a promising anti-cancer compound. Currently in Phase III clinical trials, flavopiridol is a potent inhibitor of several cyclin-dependent kinases (CDKs). Rohitukine was first reported from Amoora rohituka (0.083% dry weight) followed by that in Dysoxylum binectariferum (0.9% dry weight), both belonging to the family Meliaceae. Here, we report incredibly high yields of rohitukine (7% dry weight) in trees of D. binectariferum from the Western Ghats, India. Crude extracts of the tree were found to be highly effective against ovarian and breast cancer lines tested.     

Agrobacterium tumefaciens-mediated genetic transformation and plant regeneration from a complex tetraploid hybrid citrus rootstock

Agrobacterium-mediated genetic transformation of a tetraploid “tetrazyg” citrus rootstock selection ‘Orange #16’ [Nova mandarin (Citrus reticulata Blanco) + Hirado Buntan pummelo (Citrus grandis L. Osbeck)] × [Cleopatra mandarin (C. reticulata Blanco) + Argentine trifoliate orange (Poncirus trifoliata (L.) Raf.)] was performed. Juvenile epicotyl segments were transformed with a construct containing a bifunctional egfp–nptII fusion gene under the control of an enhanced double CaMV 35S promoter. Our protocol resulted in a reasonable transformation efficiency of 18%. Stable integration of the transgene was confirmed by visual observation of EGFP expression, PCR and Southern blot hybridization. The purpose of this work was to investigate the amenability of novel citrus rootstock germplasm being developed for improved tree size control, soil adaptation, and disease resistance, to existing transformation technologies. Seed trees of such transgenic tetraploids also have potential as trap plants containing potent insecticidal transgenes, due to their inedible fruit and inherent crossing barriers with conventional commercial diploid scion cultivars, and could be planted around producing citrus groves.     

Detection of reserpine in horses by high-performance liquid chromatography

SUMMARY A high-performance liquid chromatography (HPLC) assay was developed for the detection of reserpine. The assay was used to monitor the plasma concentrations of the drug given intramuscularly on one or two occasions to five horses. The blood concentrations of reserpine varied quite considerably between horses given the same dose of the drug. However, on average, reserpine could be detected consistently, and quantified, for 48 h after a single dose of 2.5 mg, and for a similar period after the second of two 2.5 mg doses given 13 d apart. Because of the apparently large variability in the pharmacokinetics of reserpine in horses, exact times cannot be given beyond which the drug will no longer be detectable in the plasma. However, following two doses of 2.5 mg reserpine given 13 d apart, at least 7 d must elapse after the second dose before there is no drug detectable in the plasma of most horses.     

Cryptococcosis in seven horses

The clinical, radiographic and post-mortem findings in 6 horses with cryptococcal pneumonia and one horse with an abdominal cryptococcal granuloma are described. In pulmonary cryptococcosis, the lesions were either diffuse and multiple, with bilateral lung involvement, or localised mainly to the dorsocaudal region of one lung. The cases of diffuse multiple cryptococcosis were thought to be associated with haematogenous spread of the fungus after gastrointestinal infection and dissemination from regional lymph nodes. The localised form of the disease was thought to have been associated with inhalation of cryptococci. In all cases of pulmonary cryptococcosis, encapsulated yeast-like organisms were demonstrated in Wright's-stained sediment of tracheal washes. In the horse with the abdominal granuloma, cryptococci were present in a fine needle aspirate sample. Isolates of Cryptococcus neoformans var gattii were recovered from 2 of the 5 horses in which cultures were attempted. In addition to a history of previous illness that may have predisposed to infection, most horses in this report had been in areas in which Eucalyptus camaldulensis, or the closely related E rudis, were growing. In humans, an epidemiological relationship between E camaldulensis and infection with C neoformans var gattii has been suggested. Cases of equine cryptococcosis carry a poor prognosis and treatment was not attempted in any of these cases.