Pulsed-field gel electrophoresis typing of Campylobacter fetus subsp. fetus isolated from sheep abortions in New Zealand

AIMS: To genotype Campylobacter fetus subsp. fetus isolates cultured from sheep abortions submitted to diagnostic laboratories in New Zealand during the year 2000 breeding season. To compare the types found nationally with those found in the Hawke' Bay region in 1999, and strains held in the New Zealand Reference Culture Collection, Medical Section (NZRM) from a study published in 1987.

METHODS: Campylobacter fetus subsp. fetus isolates cultured by veterinary diagnostic laboratories in the year 2000 breeding season, from sheep abortions from throughout New Zealand, were typed using pulsed-field gel electrophoresis (PFGE). In addition, seven freeze-dried C. fetus subsp. fetus isolates (strain numbers 2939–2945) from the NZRM, representing restriction types a–g found amongst sheep abortion isolates in a study published in 1987, were typed using PFGE.

RESULTS: In total, 293 C. fetus subsp. fetus isolates from 200 farms were obtained from veterinary diagnostic laboratories. Twenty-two distinct PFGE profiles were identified amongst the isolates. PFGE type B1 was predominant in each region of New Zealand and was identified from 66% of farms overall. Of the C. fetus subsp. fetus restriction types a–g lodged with the NZRM, 3/7 had PFGE profiles indistinguishable from profiles found in the current study. The other four restriction types had PFGE profiles that were unique but similar to those found in the current study.

CONCLUSIONS: PFGE type B1 was predominant amongst the C. fetus subsp. fetus isolates cultured from sheep abortions in each region of New Zealand in the year 2000, as was found in Hawke' Bay in 1999. The similarity between PFGE profiles of C. fetus subsp. fetus sheep abortion isolates from 1987 and 2000, and the relative prevalence of the PFGE groups, suggests that there has been no major genotypic shift in the population of C. fetus subsp. fetus implicated in sheep abortion in New Zealand during this time.     

Identifying factors which maximise the lambing performance of hoggets: a cross sectional study

AIM: To identity perceptions of farmers with respect to advantages of hogget lambing and to identify optimal management strategies for hogget lambing used in New Zealand.

METHODS: A questionnaire was sent to 1,038 sheep farmers in New Zealand who had indicated their hoggets would be lambing in 2002. Information was gathered on the practice of hogget lambing in New Zealand, in particular on the numbers and breeds of hoggets and rams used, selection criteria used for hoggets, use of teasers (vasectomised rams), length of mating period, use of mating crayons, vaccinations, medications and mineral supplements given, level of feed offered, use of pregnancy scanning, management during pregnancy and lambing, number of lambs present at docking, and shearing policies for hoggets. Farmers were also asked to indicate the relative importance of five stated reasons for choosing to have their hoggets mated in 2002.

RESULTS: A total of 629 (60.6%) responses were incorporated in the analysis, and the average percentage of hogget lambing was 60%. Use of vaccinations, length of the mating period, number of rams used, breed of hogget, weight at mating, management during lactation, and shearing policy were associated (p<0.05) with the lambing performance of hoggets, in the multivariate analysis.

CONCLUSIONS: This study indicated that to ensure high lambing percentages (LP) in hoggets in New Zealand, sheep farmers should: vaccinate hoggets against toxoplasmosis and campylobacteriosis; utilise a 40-day mating period; use 2.6–3.5% of rams during mating; incorporate Finn or East Friesian genetics; ensure liveweights at mating are as high as possible; manage single- and multiple-bearing/-rearing hoggets separately during the lambing and lactation periods; and shear hoggets pre-mating.     

The Effect of Parity of the Dam on Sexual Maturation,Serum Concentrations of Metabolic Hormones and the Response to Luteinizing Hormone Releasing Hormone in Bull Calves

The objectives of this study were to determine the effect of parity of the dam on age at which a scrotal circumference (SC) ≥ 28 cm was attained and the LH response to Luteinizing Hormone Releasing Hormone (LHRH) in bull calves. We also wanted to confirm, in a large group of bull calves, that the LH response to LHRH could be used to select early maturing bulls. Body weight and SC of the bull calves were measured every other week. At 15, 20 and 25 weeks of age, calves received 4.12 nm /kg body weight of LHRH ischio‐rectally and blood samples were taken every 15 min for 4 h. Calves from primiparous and multiparous dams were separated into two sub‐groups based on age at which an SC ≥ 28 cm was attained (early or late). An SC ≥ 28 cm was attained earlier in calves born to multiparous as compared with primiparous dams (p < 0.05). At 20 and 25 weeks of age, peak serum LH concentrations (LH‐peak) and area under the LH response curve (LH‐AUC) in response to LHRH were higher (p < 0.01) in calves born to multiparous as compared with primiparous dams. In calves born to multiparous dams the LH‐peak at 15 and 25 weeks of age and the LH‐AUC at 15 weeks of age were lower (p < 0.05) in calves that attained an SC ≥ 28 cm early as compared with late. The LHRH‐challenge test sensitivity and specificity ranged from 46% to 86%. We concluded that parity of the dam affected age at which SC ≥ 28 cm was attained and the LH response to LHRH in bull calves. Serum LH responses to LHRH at 15 and 25 weeks of age, in calves born to multiparous dams, show some promise for development into a test to select early maturing bull calves.     

In vitro production of porcine zygotes using intracytoplasmic injection of vitrified sperm

The objectives of this study were to evaluate if vitrified porcine spermatozoa are able to maintain their capacity to produce zygotes in vitro using intracytoplasmic sperm injection (ICSI) and to evaluate the zygote development in two in vitro atmospheric conditions: 5% CO₂ and tri‐gas. A group of porcine oocytes maturated in vitro were injected with vitrified‐warmed sperm (treatment group) and another group, with sperm diluted and conserved at 17°C (control group). To evidence parthenogenetic activation, some oocytes were submitted to a Sham test. The injected oocytes were cultured in G1 medium at 38°C, 100% humidity and 5% CO₂ or tri‐gas. No significant differences (p > .05) were observed in embryo development between the oocytes injected with vitrified‐warmed sperm (31.8%; 36/113), and those injected with semen diluted and conserved at 17°C (35.5%; 32/90), when cultured in 5% CO₂ or under tri‐gas atmosphere (42.9%; 39/91 vs. 34.2%; 26/76, respectively). No significant differences (p > .05) were observed in the percentage of pronuclei (PN) obtained between 5% CO₂ and tri‐gas, within each treatment either. Of the 52 oocytes submitted to the Sham test, only two presented a female PN (activation) indicating that the PN observed in the treatment group were a product of fertilization and not parthenogenetic activation. To conclude, porcine sperm vitrified using spheres, at a concentration of 5 × 10⁶ spermatozoa/ml in TALP medium with 1% bovine serum albumin (BSA), conserve condensed and intact chromatin capable of producing early embryo development up to the pronuclear stage.     

Closterovirus infection and mealybug exposure are necessary for the development of mealybug wilt of pineapple disease

ABSTRACT The roles of Pineapple mealybug wilt-associated viruses (PMWaVs) and mealybug (Dysmicoccus spp.) feeding in the etiology of mealybug wilt of pineapple (MWP) were evaluated. Container-grown pineapple (Ananas comosus) plants from five commercially grown Hawaiian proprietary selections and a field study utilizing a randomized complete block design were used to test four treatments for induction of MWP: PMWaV-1-free and PMWaV-1-infected plants maintained mealybug-free, and PMWaV-1-free and PMWaV-1-infected plants that received monthly applications of nonviruliferous mealybugs. A second PMWaV, PMWaV-2, was identified in some of the test plants during the course of these studies and was shown to be an integral factor in MWP etiology. Typical MWP symptoms developed only in plants infected with PMWaV-2 and exposed to mealybugs. MWP did not develop in PMWaV-1-free or PMWaV-1-infected plants that were exposed to mealybugs, or in mealy-bug-free plants infected with PMWaV-1, PMWaV-2, or both viruses. Plants from all five Hawaiian proprietary selections developed MWP when PMWaV-2 infected plants were exposed to mealybug feeding. A PMWaV-2-specific monoclonal antibody was produced that decorated the particles in immunosorbent electron microscopy and detected the virus in tissue blot immunoassays. PMWaV-2 was acquired and transmitted by pink and gray pineapple mealybugs (Dysmicoccus spp.) to pineapple plants, and these plants subsequently developed MWP symptoms while sustaining mealybug populations.     

Association Between ETFA Genotype and Activity of Superoxide Dismutase,Catalase and Glutathione Peroxidase in Cryopreserved Sperm of Holstein–Friesian Bulls

The aim of this study was to determine whether C/T missense mutation within the ETFA gene is associated with sperm antioxidant enzymatic activity. One hundred and twenty Holstein–Friesian bulls were genotyped by the PCR‐RFLP technique (MwoI). Commercial straws of frozen‐thawed semen were used to evaluate the activity of three antioxidant enzymes: superoxide dismutase, catalase and glutathione peroxidase. Among all bulls investigated, genotype CT was the most frequent (44.2%), in comparison with CC (42.5%) and TT (13.3%). Significant differences in glutathione peroxidase activity were observed between homozygous individuals (CC vs TT) with heterozygous CT having intermediate values. Dismutase activity was significantly associated with ETFA genotype, although only bulls with the CT genotype were significantly different from bulls carrying the CC genotype. The activity of catalase showed a similar trend (but was not statistically significant). In conclusion, we found that bulls with the ETFA TT genotype produce sperm with the highest glutathione peroxidase activity and can therefore be more efficiently protected from reactive oxygen. The mechanism of this interaction needs to be elucidated in future research.     

Unusual ovarian activity in a mare preceding the development of an ovarian granulosa cell tumour

An 8-year-old mare, with a foal at foot, was inseminated on foal heat with frozen semen, with the resultant pregnancy lost between days 34 and 41. The right ovary developed a large anovulatory follicle that was non-responsive to multiple doses of ovulating agents. The follicle eventually appeared to luteinise, although plasma progesterone concentrations did not reflect this. Another follicle developed, responded to GnRH and resulted in a pregnancy from frozen semen that went to term with a healthy foal. When the mare was examined after foaling, the structure on the right ovary appeared to be a granulosa cell tumour; the left ovary was smaller than normal and non-functional. Surgical removal of the right ovary before increasing photoperiod resulted in a return to function of the left ovary and a pregnancy to frozen semen on the second cycle following removal. Figures showing concentrations of inhibin, progesterone, androstenedione, oestradiol and testosterone are presented for this entire period. Unusual ovarian activity in the mare might be a prelude to the development of a granulosa cell tumour.     

An evaluation of Irish cattle herds with inconclusive serological evidence of bovine brucellosis

Since 1998, there has been a steady decline in herd restrictions and de-populations in Ireland due to bovine brucellosis. There is concern that the interpretation of laboratory results may become increasingly problematic, as brucellosis prevalence falls in Ireland. Therefore, the purpose of the current study was to evaluate the infection status of Irish herds and animals with inconclusive serological evidence of bovine brucellosis. During 12 months from September 1, 2004, laboratory and observational epidemiological data were collected from all Irish herds where animal testing identified at least one animal with a complement fixation test (CFT) reading greater than zero and/or a positive result to the indirect enzyme-linked immunosorbent assay (iELISA). Due to the observational nature of the study, we have robust estimates of the relative, but not the absolute, performance of the CFT, iELISA and brucellin skin test (BST). Herds were divided into three categories (Group A, B or C) on the basis of test results at initial assessment. A total of 639 herds were enrolled into the study, and observed for at least two years following enrolment. A rising CFT titre, with a CFT reading of 111 International CFT Units (IU) or greater at the subsequent blood test, was generally associated with herds where other evidence of infection was also available. Knowledge of the CFT reading at the initial and a subsequent blood test proved useful in distinguishing false-positive and true-positive brucellosis results. There was poor correlation between the CFT and iELISA results, and between the CFT and BST results. As a result of this study, national policy has been modified to include re-sampling of all animals with CFT readings of 20 IU or greater. This project has also led to a reduction in the number of herds restricted, as well as restriction duration. It has also contributed to a reduction in the number of herds listed for contiguous tests, and therefore the potential for contiguity testing of false positive results.