Prevalence of and Resistance to Anti‐Microbial Drugs in Selected Microbial Species Isolated from Bulk Milk Samples

The prevalence of strains of Staphylococcus aureus, coagulase‐negative (CN) staphylococci, Listeria monocytogenes, Escherichia coli, Enterococcus faecalis, E. faecium and Bacillus cereus, was investigated in 111 bulk milk samples. Staphylococcus aureus was isolated from 38 samples, CN staphylococci from 63 samples, E. coli from 49 samples, E. faecalis or E. faecium from 107 samples, and L. monocytogenes from two samples. Bacillus cereus was not found in any of the samples and three samples were free of any of the selected species. Sensitivity to the anti‐microbial drugs amikacin, ampicillin, ampicillin + sulbactam, cephalothin (CLT), cephotaxime, clindamycin, chloramphenicol (CMP), co‐trimoxazole, erythromycin (ERY), gentamicin, neomycin, norfloxacin, oxacillin, penicillin, streptomycin (STR), tetracycline (TTC) and vancomycin was tested using the standard dilution technique. Minimum inhibitory concentration (MIC) characteristics (MIC₅₀, MIC₉₀, MIC range) were determined for each microbial species. Resistance against one or more anti‐microbial drugs was found in 93% of S. aureus, 40% of CN staphylococci, 73% of E. coli, 88% of E. faecalis, 55% of E. faecium, and one L. monocytogenes strain. Most of the strains, particularly enterococci, were resistant to STR, TTC, and ERY (MIC₅₀ 4 μg/ml). A high percentage of staphylococci were resistant to β‐lactam antibiotics. High resistance to CLT was found in 11 strains of E. coli (MIC 256 μg/ml) and strains resistant to CMP (MIC₉₀ 16 μg/ml) were detected. The highest numbers of resistance phenotypes were found in E. coli (16) and CN staphylococci (12). Eighteen identical resistance phenotypes were demonstrated in indicator bacteria (E. coli, E. faecalis, E. faecium) and pathogens (S. aureus, CN staphylococci) isolated from the same bulk milk sample. The obtained resistance data were matched against the herd owners' information on therapeutic use of the drugs. This confrontation could not explain the findings of strains resistant to ERY or CMP. Our findings are evidence of selection of resistant strains among not only pathogenic agents, but also among indicator bacteria which can become significant carriers of transmissible resistance genes.     

Participatory Policy Making by Dairy Producers to Reduce Anti‐Microbial use on Farms

Pressures for more responsible use of anti‐microbial (AM) medicines in food animals are likely to increase from policymakers and the food industry, including retailers. To address this challenge, participatory approaches to welfare interventions and disease prevention may also be necessary alongside more conventional regulatory measures. This article describes the process of enabling groups of dairy producers to use a participatory policy making approach to develop an AM stewardship policy. The policy includes measures agreed to by all producers for more responsible use of AMs, whilst maintaining or improving dairy herd health and welfare. This process provided a unique opportunity for collaboration and dialogue between producers, veterinarians, industry and researchers. Its participatory nature encouraged comprehensive learning for all involved. This integration of science with producers’ knowledge and experience led to credible and practical recommendations designed to deliver real and lasting change in AM use. The multidisciplinary nature of this research marks a significant contribution to embedding social science skills and approaches into the veterinary sphere. As an initial step in creating better understanding of how participatory approaches with farmers can be applied in a UK context and more widely, this work serves as a pilot for promoting more responsible use of veterinary medicines in other livestock species.     

The Value of the Determination of Anti‐Aspergillus IgG in the Serodiagnosis of Canine Aspergillosis: Comparison with Galactomannan Detection

Diagnosis of canine aspergillosis is difficult using currently available methods. It often passes unnoticed or is diagnosed in the later phases of the disease. We developed an ELISA technique to detect anti‐Aspergillus antibodies in canine serum using an Aspergillus antigenic mycelial extract, which could then be used for the diagnosis of canine aspergillosis. We used a cut‐off of X + 3SD obtained from 20 control sera. The test was performed on 46 dogs with lesions indicating possible aspergillosis and gave nine positive results: one systemic mycosis, two discospondylitis, one uveitis, two bronchopulmonary processes and three rhinitis. We compared this methodology with the PLATELIA technique in the follow‐up of the affected dogs, obtaining the same limitations as in the diagnosis of human aspergillosis. We consider our ELISA technique using sera samples a speedy, safe and reliable method which enables us to follow up the evolution of the disease and the efficacy of the therapy chosen. A definitive diagnosis must still take into account the results of other tests such as clinical examination, radiographic studies, endoscopy and biopsy.     

Phenotypic and Genotypic Anti‐Microbial Resistance Profiles of Campylobacters from Untreated Feedlot Cattle and Their Environment

Anti‐microbial resistance is an emerging public health issue. Farmed animals may act as reservoirs and potential sources of anti‐microbial resistant Campylobacters. The aim of this study was to investigate the anti‐microbial resistance profile of cattle and environmental Campylobacter isolates from normal untreated feedlot cattle, the role of the gyrA Thr‐86‐Ile mutation in ciprofloxacin‐resistant Campylobacter jejuni isolates and the involvement of the tripartite CmeABC efflux system for multi‐resistant C. jejuni isolates. The phenotypic anti‐microbial resistance testing was carried out on 500 Campylobacter isolates (445 cattle isolates and 55 environmental isolates). In general, there was a higher level of anti‐microbial resistance for the environmental isolates compared with the animal isolates, 45% of the animal isolates were resistant to one or more of the seven anti‐microbials compared with 84% of the environmental isolates. The combined cattle and environmental Campylobacters had 34 (6.8%) isolates resistant to three or more of the seven anti‐microbials tested on all isolates and 11 (2.2%) isolates were resistant to the seven anti‐microbials. There was a substantial level of ciprofloxacin‐resistant Campylobacters in both animal (8.5%) and environmental (21.8%) isolates. The gyrA Thr‐86‐Ile mutation was only present in five of 22 ciprofloxacin‐resistant C. jejuni isolates investigated. No multi‐drug‐resistant associated mutation was detected in the CmeB or the CmeR regions investigated. In conclusion, our study observed a substantial level of Campylobacter anti‐microbial resistance, highlighting the need for an active anti‐microbial surveillance program for food animals in Ireland and the importance of the chosen sampling point can have on the findings of such a program.     

Chemical Characterization of Bovine Urine with Special Reference to Oestrus

Cows' urine was analysed by gas chromatography–mass spectrometry (GC-MS). The profiles from preovulatory, ovulatory and postovulatory samples were compared to establish any qualitative and quantitative differences that might have potential value in olfactory communication. Dichloromethane was used as the solvent for extraction of the compounds. Seven different compounds were detected, of which only two were common to all the chromatograms. The chemical profile of oestrous urine was distinguished by the presence of two specific compounds, viz. di-n-propyl phthalate and 1-iodoundecane, that were not found in the other samples. As oestrous urine has been shown to elicit sexual behaviour in cattle, these two characteristic peaks may represent important chemical compounds that elicit signals that allow the bull to detect `oestrous odours'.     

Treatment of Persistent Mating‐Induced Endometritis in Mares with the Non‐Steroid Anti‐Inflammatory Drug Vedaprofen

Recently, successful treatment of mares with a history of persistent mating‐induced endometritis (PMIE) with dexamethasone has been reported. As systemic treatment of horses with glucocorticoids should be handled with caution, we tested the hypothesis that treatment with the non‐steroid anti‐inflammatory drug (NSAID) vedaprofen, an inhibitor of cyclooxygenase‐2, may have comparative, positive effects on fertility. Barren mares with a history of repeated PMIE were treated with vedaprofen (n = 8; initially 2 mg/kg bodyweight followed by 1 mg/kg orally twice daily) from 1 day before the first insemination to 1 day after ovulation or left untreated (n = 9). All mares received oxytocin (20 I.E. s.c.) thrice daily. Uterine swabs were collected for bacteriology and cytology. The day after ovulation, fluid accumulation was detected in three control mares and four treated mares (n.s.). The percentage of neutrophils in uterine cytology was significantly increased in comparison to the day before ovulation irrespective of treatment. Pregnancy was confirmed in two of nine mares in the control group and seven of eight mares in the treatment group (p < 0.05). NSAIDs may positively affect fertility in mares with a history of PMIE.     

Ultrastructure of Rabbit Embryos Exposed to Hyperthermia and Anti‐Hsp 70

The aim of the study was to determine the effect of short‐term hyperthermia and Hsp70 blockage on ultrastructural changes in cell organelles and nucleoli of rabbit preimplantation embryos. The embryos were cultured either at 37.5°C (control, C) or 41.5°C (hyperthermia, HT) during 6 h. The antibody against Hsp70 was added into the culture medium (4 μg/ml) of morula stage embryos from C and HT groups. After termination of the culture, the embryos were processed for transmission electron microscopy. The embryos exposed to hyperthermia showed increased volume of lipid droplets, considerable occurrence of cellular debris in the perivitelline space and slight changes in the occurrence of microvilli on the surface of trophoblastic cells. In the embryos exposed to anti‐Hsp 70 at 37.5°C, there were considerable changes in mitochondria morphology, decreased volume of dense bodies in the cytoplasm and considerable changes in the occurrence of microvilli on the surface of trophoblastic cells. In the group of embryos exposed simultaneously to hyperthermia and anti‐Hsp 70, mitochondria were also expanded and swollen; the volume of flocculent vesicles and lipid droplets was increased and the volume of dense bodies in the cytoplasm was diminished. General organization of the cytoplasm in groups with anti‐Hsp70 was characterized by cell organelle segregation. Averaged size of the nucleolar area was significantly increased in the embryos exposed to hyperthermia, whereas in the group exposed to the anti‐Hsp70 without hyperthermia it was significantly diminished. Hyperthermia also caused disintegration of compact status of the nucleoli. In presence of anti‐Hsp 70, the structural changes, described within the nucleoli during hyperthermia, were not observed. In conclusion, these results document ultrastructural changes in cell organelles of rabbit preimplantation embryo caused by hyperthermia, and also changes in the nucleolar structures, at which presence of Hsp‐70 inhibit these changes.     

Systematic Review of Nonsteroidal Anti‐Inflammatory Drug‐Induced Adverse Effects in Dogs

The aim of this systematic review was to identify, assess, and critically evaluate the quality of evidence of nonsteroidal anti‐inflammatory drug (NSAID)‐induced adverse effects in dogs. Original prospective studies published in peer‐reviewed journals in English (1990–2012) that reported data on the safety of NSAIDs administration in dogs were searched. For each study, design type (I, II, III, or IV) and assessment of quality (+, Ø, ?) were rated. For each drug, quantity and consistency rating (***, **, *) and strength of evidence (high, moderate, low, or extremely low) were identified and evaluated. The strength of evidence was defined in terms of how applicable and relevant the conclusions were to the target population. Sixty‐four studies met the inclusion criteria. Thirty‐five (55%) research studies and 29 (45%) clinical trials were identified. A high strength of evidence existed for carprofen, firocoxib, and meloxicam; moderate for deracoxib, ketoprofen, and robenacoxib; and low for etodolac. Quality and consistency rating were as follows: carprofen (***/***), deracoxib (**/***), etodolac (*/unable to rate), firocoxib (***/**), ketoprofen (**/***), meloxicam (***/***), and robenacoxib (**/**), respectively. Adverse effects were detected in 35 studies (55%) and commonly included vomiting, diarrhea, and anorexia. Three studies (5%) reported a power analysis related to adverse effects of ≥80%. In randomized, placebo‐controlled, blinded studies (n = 25, 39%), the incidence of adverse effects was not statistically different between treated and control dogs. Finally, most studies were not appropriately designed to determine the safety of NSAIDs, and involved a healthy nongeriatric population of research dogs.     

Anti‐Müllerian Hormone in the Domestic Dog during the Anestrus to Oestrous Transition

The reproductive cycle of the domestic dog features a long period of relative ovarian inactivity or anestrus. The mechanism of anestrous termination/oestrous resumption is not yet fully understood, which presents a challenge to the development of oestrous induction protocols. In this study, we assess the possibility that anti‐Müllerian hormone (AMH) might play a role in this transition by characterizing its patterns of expression in the circulation during the transition from anestrus to oestrous and in all stages of ovarian follicular growth. Serum samples from five beagles (2.0–4.5 years) were collected three times per week at least 30 days prior to the onset of oestrous and assessed for AMH concentrations. Serum AMH concentration increased significantly during the transition from anestrus to proestrus and then declined back to the anestrous baseline beginning on day ?4 before the luteinizing hormone surge, which was determined by changes in serum progesterone concentrations. Cortical sections of ovaries from females undergoing routine ovariohysterectomy (aged 8 months–5 years, n = 4) were evaluated for AMH by immunohistochemistry. Pre‐antral and small antral follicles were most strongly immunoreactive for AMH. These data suggest that the increase in the number of antral follicles is associated with the rise in serum AMH as the anestrous period comes to an end. The rise in AMH might be useful in predicting the onset of oestrus and therefore assist with the optimization of oestrous induction protocols and possibly other assisted reproductive technologies.     

β-Lactoglobulin Phenotypes in Finnish Ayrshire and Friesian Cattle with Special Reference to Mastitis Indicators

An unselected material consisting of composite milk samples from 1029 Ayrshire and 113 Friesian cows were analysed for the β-lactoglobulin types. The frequencies of β-Lg types were AA 8.3 %, AB 45.5% and BB 46.3 % for Ayrshire cows and 22.1 %, 45.1 % and 32.8 % for Friesian cows, respectively. The relationship between β-Lg type with milk BSA, somatic cell count, protein percentage, fat percentage and milk production were analysed. AA cows were significantly higher in daily milk yield but lower in protein percentage and fat percentage than BB cows; AB types were intermediate. The annual production was highest in AB-animals. There was a tendency for AA cows to have high somatic cell counts but low milk BSA concentrations.β-lactoglobulins; milk BSA; somatic cell count; bovine mastitis; bovine milk; milk proteins.     

Meta‐Analyses of Factors Associated with Leptospirosis in Domestic Dogs

Factors related with leptospirosis in domestic dogs have been reported worldwide. The aims of this study were to identify factors associated with this disease described in different observational studies and to combine the coinciding factors in at least four studies using meta‐analyses, to obtain a pooled odds ratio (OR) as measure of infection risk. A literature search was performed in electronic databases, electronic databases of specific journals and search engines to find studies published in English, Spanish and Portuguese available from January 1960 to January 2015. Two hundred and eighteen factors were identified in 31 publications including cross‐sectional and case–control studies. Finally, independent meta‐analyses were performed with six different variables, which included between 4 and 8 articles. The pooled OR indicated that the variables ‘mixed‐breed dogs’, ‘flooding occurrence in the habitat of the dog’ and ‘working dogs’ were risk factors for leptospirosis, while ‘being a dog less than 1 year old’ was a protective factor; however, all these associations were not statistically significant. Otherwise, the variables ‘male dog’ and ‘urban dog’ were statistically significant risk factors for infection. This study highlights the need for more formal studies on the epidemiology of canine leptospirosis. Nevertheless, the study revealed that some risk factors for infection coincided in different observational studies. These factors could be considered to raise suspicion about the disease, especially when there is a history of exposure to the bacteria.     

Anti‐Muellerian hormone concentration in bitches with histopathologically diagnosed ovarian tumours and cysts

Increased concentrations of Anti‐Muellerian hormone (AMH) can indicate a granulosa cell tumour as shown in women, mares and cows. To investigate AMH to differentiate canine granulosa cell tumour from other ovarian pathologies, we evaluated the ovaries of 63 bitches. Blood serum samples were collected before surgery for AMH analysis. Ovaries were submitted for histopathological examination. Fourteen bitches showed normal ovaries. These bitches had AMH values between 0.12 and 0.99 ng/ml. In 20 bitches ovarian cysts i.e., follicular cysts (n = 8), corpora lutea cysts (n = 7), subsurface cysts (n = 5) were diagnosed. These dogs had AMH values of 0.11–2.09 ng/ml. Bitches with small luteinized follicular cysts had slightly higher AMH values than those without ovarian alteration. In 29 cases ovarian neoplasms i.e., granulosa cell tumour (n = 9), epithelial tumours (n = 16), dysgerminomas (n = 3) and one sarcoma were identified. Anti‐Muellerian hormone values of bitches with an ovarian neoplasm except granulosa cell tumour ranged from 0.18 to 1.18 ng/ml. The AMH values of bitches with granulosa cell tumour ranged from 1.12 to ≤23 ng/ml and were significantly higher (p < .05) than in all of the other bitches. The cut‐off of 0.99 ng/ml gave a sensitivity of 100% and a specificity of 94.44% to diagnose granulosa cell tumour. In conclusion, markedly elevated AMH concentrations in bitches are indicative for a granulosa cell tumour. However, negative testing does not rule out the existence of small one. Differentiation of GCT from luteinized follicular cysts may especially be difficult.