Prevalence and clonal diversity of Campylobacter jejuni from dairy farms and urban sources

AIM: To investigate the role of free-living animals such as spar- rows, rodents and flies as potential reservoirs of Campylobacter spp on a dairy farm, and to assess the genetic diversity among Campylobacter isolates from the farm and an urban source.

METHODS: A total of 290 samples (bovine, passerine and ro- dent faeces, and whole flies) were collected from a large com- mercial dairy farm in the Manawatu district in New Zealand, and from faeces from urban sparrows in a nearby city. Other samples collected from the dairy farm included five from silage, two from aprons worn by workers during milking, two from workers' boots and two from water in troughs in a paddock. Isolates of thermophilic Campylobacter spp were identified mor- phologically and phenotypically and further characterised mo- lecularly using pulsed-field gel electrophoresis (PFGE) and the restriction enzyme SmaI.

RESULTS: Campylobacter jejuni was the only Campylobacter species isolated from all samples. The highest prevalence was found in faeces from dairy cows (54%), followed by faeces from sparrows from the urban area (40%) and the farm (38%), and from rodents (11%) and whole flies (9%). Other samples from the farm environment such as silage, trough water, and work- ers' aprons and boots were also positive for C. jejuni. Of the 22 restriction patterns obtained, seven were common to more than one source.

CONCLUSIONS: Cattle, sparrows, rodents and flies are po- tential reservoirs of C. jejuni on dairy farms. Identical clones of C. jejuni carried by cattle, sparrows, flies and rodents prob- ably indicate a common source of infection. The high level of asymptomatic carriage of C. jejuni by healthy dairy cows could be sufficient to maintain infections within the dairy farm sur- roundings via environmental contamination.     

The effect of the maturity and prior breeding activity of rams and body condition score of ewe hoggets on the reproductive performance of ewe hoggets

Abstract

AIM: To determine the effect of age and prior use of mature rams at a given ram-to-ewe ratio, and the effect of body condition on breeding performance and pregnancy rate of ewe hoggets.

METHODS: Ewe hoggets (n=733) aged 7–8 months were weighed and their body condition scored, then randomly assigned to one of three treatment groups (Day 0) and joined with either four two-tooth rams (20 months of age) not used previously (n=244; Two-tooth), four mixed-aged mature rams that had not been used earlier in the season (n=244; Mature-fresh), or four mixed-aged mature rams that had been used with mature ewes immediately prior to joining with hoggets (n=245; Mature-used). The breeding period was 34 days. Ewe hoggets were identified as having been marked during the first 17 days only, during both 17-day periods, during the second 17 days only, or not marked. Hoggets were re-weighed on Day 34, and pregnancy status determined using ultrasound on Day 92. The breeding soundness of the rams was assessed on Days ?34 and ?1.

RESULTS: Semen samples obtained from the rams did not differ significantly in any of the parameters measured (p>0.05). Ewe hoggets joined with Mature-fresh rams were less likely (p<0.05) to be marked in the second 17 days of breeding only than those joined with either Two-tooth or Mature-used rams. No other breeding parameters were affected by breeding group (p>0.05). Hoggets marked in the first 17 days only were heavier (p<0.05) at Day 0 than those marked in the second 17 days only or not marked. Hoggets diagnosed as twin-bearing were heavier (p<0.05) than non-pregnant or single-bearing hoggets. Those hoggets marked in the first 17 days only had a greater (p<0.05) body condition score (BCS) than those marked in the second 17 days only or not marked at all. These differences were no longer apparent after correction for liveweight (LW).Correction for LW at Day 0 or change in LW during the breeding period did not affect the results for breeding performance.

CONCLUSIONS: Under the conditions of this study, two-tooth rams and mature rams that had been used previously were just as suitable as mature rams that had not been used previously for breeding with ewe hoggets. Further studies are warranted to verify this result. The re-use of rams without reducing breeding performance would reduce breeding costs and may make breeding hoggets a more viable option for farmers. The BCS of ewe hoggets affected breeding performance, and can thus be used to identify those animals most suitable for breeding.     

Comparison of concentrations of copper in plasma and serum from farmed red deer (Cervus elaphus)

AIM: To determine the relationship between the concentrations of Cu in plasma and serum in red deer, and to compare this relationship with those previously reported in cattle and sheep.

METHODS: Paired serum and heparinised plasma samples from 114 red deer from 10 herds (n=6-20 per herd) were analysed for concentrations of Cu. Samples were collected either at slaughter (n=84; eight herds) or by jugular venepuncture (n=30; two herds). Thirty-nine of the samples taken at slaughter were from adult hinds from four herds, while other samples were taken from 10–14-month-old males, except for one herd (10 samples) where an equal number of 8–9-month-old males and females were sampled. The effect of age, gender and herd on the relationship between concentrations of Cu in plasma and serum was assessed using univariate ANOVA. The individual results for concentrations of Cu in serum were compared with those in plasma, using limits-of-agreement plotting.

RESULTS: The mean concentration of Cu in plasma was not significantly different from that of serum (0.048; 95% CI=-0.14 to 0.24 µmol/L). There was no effect of age, sex or herd on this relationship.

CONCLUSIONS: In deer, there was no significant difference between concentrations of Cu in plasma and serum regardless of age, sex or herd of origin.

CLINICAL RELEVANCE: In contrast to the situation in cattle and sheep, the concentration of Cu in serum can be used interchangeably with that in plasma for the estimation of concentration of Cu in blood of red deer.     

Effects of supplementation of medium with different antioxidants during in vitro maturation of bovine oocytes on subsequent embryo production

The aim of this study was to assess the effects of different antioxidants on the levels of reactive oxygen species (ROS) and glutathione (GSH) in oocytes during in vitro maturation (IVM), as well as on the production of embryos. Oocyte of slaughterhouse‐derived cattle ovaries were placed in IVM with different antioxidants: quercetin (2 μM), cysteamine (100 μM), carnitine (0.5 mg/ml), vitamin C (50 μg/ml) or resveratrol (2 μM). Oocytes matured without any antioxidant supplementation were used as control. The oocytes were assessed for maturation rates and for ROS and GSH levels by fluorescence staining in 2′,7′‐dichlorodihydrofluorescein diacetate and Cell Tracker Blue, respectively. Embryo production was assessed in terms of cleavage, blastocysts and hatching rates and embryo cell numbers. The results expressed in arbitrary fluorescence units showed ROS reduction (p < .05) in the groups with quercetin (27.5 ± 3.4), vitamin C (27.1 ± 3.0) or resveratrol (28.1 ± 4.7), in comparison with those with cysteamine (34.9 ± 4.5), carnitine (34.6 ± 3.8) or to the control group (36.5 ± 5.2). GSH levels increased (p < .05) in cysteamine (63.5 ± 5.5) or carnitine (60.8 ± 4.4) groups in comparison with quercetin (52.7 ± 5.1), vitamin C (53.0 ± 3.8), resveratrol (53.1 ± 4.4) or to the control (49.6 ± 4.5). Nuclear maturation cleavage and hatched blastocysts rates did not differ (p > .05) between groups. However, blastocyst rates after in vitro fertilization in quercetin (53.5 ± 3.9%), vitamin C (52.1 ± 3.1%) resveratrol (54.2 ± 4.0%), cysteamine (52.4 ± 2.7%) or carnitine (54.2 ± 3.1%) groups were higher (p < .05) than in the control (47.2 ± 2.7%). Total cell numbers in embryos from the vitamin C, resveratrol, cysteamine or carnitine groups were higher than in quercetin and control groups, which were similar to each other. The results suggest that using antioxidants during IVM may reduce oxidative stress either by decreasing ROS levels directly or by increasing GSH levels in oocytes, depending on the type of antioxidant used. Overall, oxidative stress control during IVM with the antioxidants examined here improved blastocyst development with similar efficacy.     

Consequences of Nitric Oxide Synthase Inhibition During Bovine Oocyte Maturation on Meiosis and Embryo Development

The importance of nitric oxide synthase (NOS) in bovine oocyte maturation was investigated. Oocytes were in vitro matured with the NOS inhibitor N^w- l -nitro-arginine methyl-ester (10⁻⁷, 10⁻⁵ and 10⁻³  m l -NAME) and metaphase II (MII) rates and embryo development and quality were assessed. The effect of l -NAME (10⁻⁷  m ) during pre-maturation and/or maturation on embryo development and quality was also assessed. l -NAME decreased MII rates (78–82%, p < 0.05) when compared with controls without l -NAME (96%). Cleavage (77–88%, p > 0.05), Day 7 blastocyst rates (34–42%, p > 0.05) and total cell numbers in blastocysts were similar for all groups (146–171 cells, p > 0.05). Day 8 blastocyst TUNEL positive cells (3–4 cells) increased with l -NAME treatment (p < 0.05). For oocytes cultured with l -NAME during pre-maturation and/or maturation, Day 8 blastocyst development (26–34%) and Day 9 hatching rates (15–22%) were similar (p > 0.05) to controls pre-matured and matured without NOS inhibition (33 and 18%, respectively), while total cell numbers (Day 9 hatched blastocysts) increased (264–324 cells, p < 0.05) when compared with the controls (191 cells). TUNEL positive cells increased when NOS was inhibited only during the maturation period (8 cells, p < 0.05) when compared with the other groups (3–4 cells). NO may be involved in meiosis progression to MII and its deficiency during maturation increases apoptosis in embryos produced in vitro . Nitric oxide synthase inhibition during pre-maturation and/or maturation affects embryo quality.     

Cloning and sequencing of the ovine gamma-interferon gene

Cytokines are major modulators of the immune system of all animals. The cloning and expression of recombinant cytokine genes have permitted the analysis of their immune function and role in the control of the immune response to disease and vaccination. While human, murine, and bovine genes have been cloned and sequenced, the cloning of ovine cytokine genes has not yet been reported. As sheep are of dominant economic importance to the Australian farming industry, it is of significance to clone and express these genes to facilitate the development of new and better vaccines and pharmaceuticals. We have initially selected ovine gamma-interferon (gamma-IFN) as a target cytokine gene. By the use of the polymerase chain reaction (PCR), using primers based on the bovine gamma-interferon sequence, we have amplified the ovine gamma-interferon gene from crude messenger RNA extracted from lymphocytes. After cloning and DNA sequencing the gene, we found that ovine gamma-IFN is 93% identical to bovine gamma-IFN in amino acid sequence. This result indicates that the PCR method will be a rapid and efficient means for cloning other ovine cytokine genes.     

Sperm Morphology Assessment in Captive Neotropical Primates

The main objective of this study was to evaluate sperm morphology in four neotropical primate species to compare the sperm morphological traits and the sperm morphometric parameters as a basis for establishing normative sperm standards for each species. Data from 80 ejaculates collected from four primate species, Callithrix jacchus, Callimico goeldii, Alouatta caraya and Ateles geoffroyi, were analysed for detection of sperm morphological alterations using subjective World Health Organization (WHO‐2010) standards and Sperm Deformity Index (SDI) criteria, objective computer‐assisted sperm morphometry analysis (CASMA) and subpopulation sperm determination (SSD) methods. There were multiple differences (p < 0.01) observed among primate species in values obtained from WHO‐2010, SDI, CASMA and SSD sperm analysis methods. In addition, multiple significant positive and negative correlations were observed between the sperm morphological traits (SDI, Sperm Deformity Index Head Defects, Sperm Deformity Index Midpiece Defects, Sperm Deformity Index Tail Defects, Normal Sperm, Head Defects, Midpiece Defects and Tail Defects) and the sperm morphometric parameters (SSD, Area (A), Perimeter (P), Length (L), Width (W), Ellipticity, Elongation and Rugosity) (p ≤ 0.046). In conclusion, our findings using different evaluation methods indicate that pronounced sperm morphological variation exists among these four neotropical primate species. Because of the strong relationship observed among morphological and morphometric parameters, these results suggest that application of objective analysis methods could substantially improve the reliability of comparative studies and help to establish valid normative sperm values for neotropical primates.