Isolation and molecular characterisation of Neospora caninum in cattle in New Zealand

AIM: To isolate Neospora caninum from the brains of naturally infected cattle and use molecular techniques to characterise the isolates.

METHODS: Neospora caninum tachyzoites were isolated in Vero cell culture from the brains of a cow and two calves. The isolates were characterised using polymerase chain reaction (PCR) methods, DNA sequencing, an immunofluorescent anti-body test (IFAT), transmission electron microscopy (TEM), and immunohistochemistry (IHC). The brains of the three cattle were subjected to histopathological examination. A pathogenicity study was conducted in 120 BALB/c mice.

RESULTS: Neospora caninum tachyzoites were isolated from all three cases and first observed in vitro between 14 and 17 days post-inoculation. Parasites were sub-cultured and maintained in Vero cell culture for more than 6 months. PCR products were generated for all three isolates, using two different primers. Sequencing of the PCR products and a subsequent BLAST search identified the isolates as N. caninum. In addition, the isolates tested positive using IFAT and IHC, and ultrastructure revealed by TEM was characteristic of N. caninum. Histopathological examination revealed lesions characteristic of N. caninum in 1/3 brains. In the pathogenicity study using BALB/c mice, the mortality rate was 3–7%.

CONCLUSION: This was the first successful isolation of N. caninum in New Zealand confirmed using molecular characterisation tests.     

The effect of social isolation,gender and familiarity with the experimental procedure on tests of porcine nociceptive thresholds

ObjectiveTo investigate the effects of habituation and isolation on mechanical nociceptive thresholds in pigs at the pelvic limbs and at the tail.Study designProspective randomized multifactorial study.AnimalsThirty-two healthy castrated male (experiment 1), and 12 castrated male and 12 female (experiment 2) Danish Landrace × Yorkshire pigs, weighing 63.5 ± 0.8 kg and 55.4 ± 0.6 kg (the mean ± SD, experiment 1 and 2, respectively).MethodsMechanical nociceptive thresholds were quantified with a von Frey anesthesiometer applied to two distinct anatomical regions (tail and pelvic limbs). Pigs receiving the mechanical challenge in the pelvic limbs were tested inside a cage, whereas pigs exposed to stimuli at the tail region were tested in an open arena. For both experiments, the effect of familiarity to the procedure was evaluated by comparing thresholds of nociception in habituated versus naïve pigs. The presence of a companion animal was also evaluated in pigs receiving stimuli at the pelvic limbs.ResultsPigs tested inside the cage were affected by the habituation to the procedure as indicated by the increase in willingness and time spent by the animals in the test cage. This effect was reflected in the lower mechanical nociceptive thresholds (medians with 25–75 percentiles) recorded for familiar pigs compared with naïve animals [495 g (302–675) versus 745 g (479–1000), respectively; p = 0.026]. Mechanical nociceptive thresholds measured at the tail of the pigs in the open arena were not affected by the familiarity of the animals with the experimental procedure.Conclusions and clinical relevanceThe current results reiterate the value of habituation in research involving animal behaviour. Further characterization of the methodology is needed to allow its application in the evaluation of clinical conditions in pigs.     

Mustelidae are natural hosts of Staphylococcus delphini group A

According to the current taxonomy, the Staphylococcus intermedius group (SIG) comprises of at least three distinct species. While S. intermedius and S. pseudintermedius are associated with specific hosts (pigeons and dogs, respectively), the natural host of S. delphini remains unclear. We analysed 158 SIG isolates from less studied animal species belonging to the order Carnivora, including mink (n=118), fox (n=33), badger (n=6) and ferret (n=1). Species identification was performed by nuc PCR in combination with sodA sequence analysis and pta PCR restriction fragment length polymorphism (RFLP). The results showed a consistent association between host and bacterial species. All isolates from minks, ferret and badgers belonged to S. delphini group A, whereas all fox isolates except one were identified as S. pseudintermedius. The remaining fox isolate belonged to S. delphini group A. The results indicate that Mustelidae such as minks, ferrets and badgers are natural hosts of S. delphini group A. This is in contrast with Canidae, which are primarily colonized and infected with S. pseudintermedius. These findings suggest that coagulase-positive staphylococcal species may have evolved and diverged through host adaptation.     

Dormant Season Grazing: Effect of Supplementation Strategies on Heifer Resource Utilization and Vegetation Use

Grazing dormant forage under low-input heifer development strategies typically exposes cattle to low-quality forage. Protein supplementation while grazing dormant range can enhance heifer growth and reproductive performance. We examined resource utilization of heifers and the effects of dormant season grazing on residual vegetation characteristics under two supplementation management strategies. Approximately 100 weaned composite heifer calves were randomly selected and placed into one of two supplementation treatments in each of 2 yr, one receiving a free access 62% crude protein self-fed concentrate and the other receiving a daily hand-fed 20% crude protein cake. Grazing occurred from December (2015 and 2016) through March (2016 and 2017). Thirty transects were randomly located within each pasture for measuring vegetation quality and structure before and after grazing. Daily space use and behavior was evaluated for 21 individuals within each treatment using global positioning system (GPS) collars and resource utilization functions. Heifers supplemented with concentrated protein spent more time grazing per day than heifers supplemented with cake (6.92 ± 0.18, 6.24 ± 0.17 h). Relative use by heifers in the cake treatment was negatively related to horizontal distance from the supplement delivery site early to midwinter ($\stackrel{?}{\overline{\beta }}$ = ? 0.41 ± 0.16, ? 0.53 ± 0.17). Both treatments selected grazing locations relative to standing biomass of perennial grasses ($\stackrel{?}{\overline{\beta }}$ = 0.0005 ± 0.00004) and crude protein ($\stackrel{?}{\overline{\beta }}$ = 0.12 ± 0.007). However, resource selection was highly variable among individuals for both supplementation treatments. We found no treatment effects on pre-post grazing differences in residual cover of litter, grass, forbs and shrubs (P > 0.24). However, the time period when grazing occurred had an effect on residual vegetation conditions (P < 0.01). Our results indicate high levels of variability in grazing site selection by heifers, suggesting future research should incorporate individual animal measurements in an attempt to account for individual animal variability.     

Golden perspective: application of laser-generated gold nanoparticle conjugates in reproductive biology

The current demand for female calves has grown rapidly and controlling the sex of offspring provides an economically flexible management for the livestock producer. The only functioning method of efficiently producing separate populations of X and Y sperm in mammals is based on relative DNA differentiation by high-speed flow cytometry. In this context, gold nanoparticles conjugated to sex chromosome-specific moieties display promising application as novel fluorophor-alternative for the high-throughput screening, since they feature no photo bleaching, high quantum yield, good biocompatibility and the possibility of non-destructive membrane penetration. Especially, gold nanoparticles fabricated by pulsed laser ablation are in the recent focus of interest, due to excellent biocompatibility, fabrication-dependent, tuneable particle size as well as surface charge and ease of (bio)-functionalization with a remarkably strong ligand binding. For the purpose of our studies functionalized gold nanoparticles may be used as novel markers for sex-sorting of mammalian sperm and, depending on the selected probe, also for the selection of sperm with heritable DNA-sequences interesting for animal breeding.     

Cross-sectional studies of ultimate pH in lambs

The ultimate pH of the longissimus muscle was measured in 1536 lambs routinely slaughtered at a meat export works in the southern part of the North Island during the 1981/82 season. The mean ultimate pH of all samples was 5.60, and 7.2 percent of the carcases had values equal to, or above, 6.00 whereas 85.3 percent of carcases had values below 5.80 which is considered to be optimal. The ultimate pH values of samples from lambs slaughtered during the summer period was significantly higher than those obtained during three other seasonal sampling periods and Perendale lambs had significantly higher ultimate pH values as compared to lambs of other breeds. There was no statistical association between distance travelled before slaughter and the ultimate pH of carcases but there was a highly significant direct correlation between holding periods of lambs in the stockyards and ultimate pH of their meat. There was also a highly significant inverse correlation between fleece weight and ultimate pH and it is suggested that both this effect and the seasonal pattern of ultimate pH values indirectly reflect the major role that nutrition may play in the development of high ultimate pH meat in lambs. It is further suggested that washing of animals prior to slaughter and the length of subsequent resting periods are important factors in relation to the development of undesirably high ultimate pH values.     

The influence of feeding crimped kernel maize silage on broiler production,nutrient digestibility and meat quality

1. Two experiments were carried out in parallel with male Ross 308 broilers over 37 d. An experiment with a total of 736 broilers was performed to study the effect of dietary inclusion of crimped kernel maize silage (CKMS) on broiler production and meat quality. Another study with 32 broilers was carried out from 21 to 25 d to investigate the inclusion of CKMS on nutrient digestibility.

2. In both trials, 4 dietary treatments were used: wheat-based feed (WBF), maize-based feed (MBF), maize-based feed supplemented with 15% CKMS (CKMS-15) and maize-based feed supplemented with 30% CKMS (CKMS-30).

3. Compared with MBF, the dry matter (DM) intakes of broilers receiving CKMS-15 and CKMS-30, respectively, were numerically 7.5 and 6.2% higher and feed conversion ratio 6 and 12% poorer (significant for 30% CKMS), although there were no significant differences in AME content between the three diets. At 37 d, the body weight of birds receiving 15% CKMS was similar to birds fed with MBF. However, the inclusion of 30% CKMS decreased broiler growth. Dietary supplementation with CKMS significantly reduced the apparent digestibility of phosphorus. The fat digestibility was significantly lower for CKMS-30 than for the other three diets.

4. Broiler mortality decreased significantly when CKMS was added to the diet.

5. The consumption of drinking water was significantly lower in all maize-based diets as compared to WBF and was lowest in broilers fed with CKMS-30.

6. An improved litter quality in terms of DM content and a lower frequency of foot pad lesions was observed with broilers supplemented with both dietary levels of CKMS.

7. The addition of CKMS to maize-based diets increased juiciness, tenderness and crumbliness of the meat.

8. In conclusion, the dietary supplementation of 15% CKMS had no negative effect on broiler growth and positively influenced bird welfare in terms of mortality and foot pad health. Therefore, the addition of 15% CKMS to maize-based diets is considered an advantageous feeding strategy in broiler production.