Leukotrienes Affect Secretory Function of Ovarian Cells In Vitro*

The aim of this study was to determine which cells are the source of production and target for leukotriene (LTs) action within the bovine ovary. Luteal (CL, days 14–16 of the oestrous cycle), steroidogenic cells (LSC) and endothelial cells (LEC) of the bovine corpus luteum (CL), and granulosa cells (GC) were isolated enzymatically, cultured in a monolayer and incubated with LTC₄, LTB₄, Azelastine (an antagonist of LTC₄) or Dapsone (an antagonist of LTB₄). Then cells were collected for determination of mRNA expression for LT receptors (LTRs) and 5‐lipoxygenase (5‐LO) by real time RT‐PCR, and media were collected for determination of prostaglandin (PG)E₂, F_2α, progesterone (P4; LSC only), endothelin‐1 (ET‐1; LEC only) and 17‐β oestradiol (E2; GC only). The greatest mRNA expression for LTR‐II and 5‐LO were found in LEC, whereas LTR‐I mRNA expression did not differ among cell types. The level of PGE₂ increased after LTs treatment in each type of ovarian cell, excluding LTC₄ treatment in LEC. The secretion of PGF_2α was also increased by LTs, but decreased after LTB₄ treatment of LSC. In GC cultures, both LTs stimulated E2 secretion; in LEC cultures, LTB₄ stimulated whereas LTC₄ inhibited P4 secretion; in LEC cultures, LTC₄ stimulated but LTB₄ inhibited ET‐1 secretion. The results show that LTs are produced locally and are involved in PGs production/secretion in all examined cells (LSC, LEC and GC) of bovine ovary. Leukotriene treatment modulate secretion of E2, by GC, P4 by LSC and ET‐1 by LEC, which indicates that LTs are involved in regulation of ovarian secretory functions.     

Candidate gene and genome-wide association studies of Mycobacterium avium subsp. paratuberculosis infection in cattle and sheep: a review

Paratuberculosis (Johne's disease), caused by Mycobacterium avium subspecies paratuberculosis, is responsible for significant economic losses in livestock industries worldwide. This organism is also of public health concern due to an unconfirmed link to Crohn's disease. Susceptibility to paratuberculosis has been suggested to have a genetic component. In livestock, a number of candidate genes have been studied, selected on their association to susceptibility in other mycobacterial diseases, their known role in disease pathogenesis or links to susceptibility of humans to Crohn's disease. These genes include solute carrier family 11 member 1 (SLC11A1, formerly NRAMP1), toll-like receptors, caspase associated recruitment domain 15 (CARD15, formerly NOD2), major histocompatibility complex (MHC) and cytokines (interleukin-10 and interferon-gamma) and their receptors. Genome wide association studies have attempted to confirm associations found and identify new genes involved in pathogenesis and susceptibility. There are a number of limitations and difficulties in these approaches, some peculiar to paratuberculosis but others generally applicable to identification of genetic associations for complex traits. The technical approaches and available information for paratuberculosis have expanded rapidly, particularly relating to sheep and cattle. Here we review the current published evidence for a genetic association with paratuberculosis susceptibility, technological advances that have progressed the field and potential avenues for future research.     

The interleukin 10 response in ovine Johne's disease

Johne's disease is an enteric mycobacterial infection of ruminants that has significant global economic impact. The classic host reaction is one of an early T-cell mediate immune response, with predominant interferon gamma (IFNγ) activity; there is subsequent lowering of this response as animals reach the terminal stages of disease. Interleukin (IL)-10, which can suppress Th1-type and enhance Th2-type cytokine production, is considered to play a role in the later stages of Johne's disease. To determine the role of IL-10 throughout the course of Johne's disease we studied groups of sheep with either no Johne's disease (n = 10), natural infection (n = 30) or experimental infection (n = 58). Disease status of the animals was comprehensively assessed by culture of Mycobacterium avium subsp. paratuberculosis (Mptb), histopathology and serology. Antigen-specific IL-10 secretion in peripheral blood of sheep exposed to Mptb was significantly higher than in control animals (P < 0.001) as early as 4 months post-inoculation, and increased progressively. In ileal and jejunal lymph node cells, IL-10 secretion was also significantly higher in animals that were exposed to Mptb compared to controls (P < 0.05). The early IL-10 response seen in peripheral blood cells may be a reflection of early responses at sites of Mptb infection. IL-10 secretion from ileal and jejunal lymph node cells was significantly higher in exposed animals with no lesions or with paucibacillary lesions when compared to animals with multibacillary lesions. These novel findings demonstrate that increased IL-10 activity commences soon after exposure to the causative mycobacterium and may play a role in determining disease outcome.     

Serological characterisation of Haemophilus parasuis isolates from Australian pigs

A total of 31 isolates of Haemophilus parasuis obtained from Australian pigs were serotyped by the Kielstein-Rapp-Gabrielson scheme. The isolates were assigned to serovar 1 (1 isolate), serovar 2 (1 isolate), serovar 4 (4 isolates), serovar 5 (7 isolates), serovar 9 (2 isolates), serovar 10/7 (4 isolates), serovar 12 (1 isolate) and serovar 13 (6 isolates). The remaining 5 isolates could not be assigned to a serovar. Two different serovars (5 and 13) were detected in one herd. The only 2 isolates obtained from clinically normal pigs (from the same herd) were serovar 9. The common serovars were isolated from pigs with pneumonia as well as from pigs with conditions of the Glässer's disease type. The serological heterogeneity amongst Australian isolates of H parasuis has important implications for the use of vaccines to control Glässer's disease.     

A serological survey to determine the prevalence of infection with Treponema hyodysenteriae in Western Australia

A serological survey to detect antibody titres against Treponema hyodysenteriae was conducted on pigs from 106 herds in Western Australia. Titres indicating a positive result in the tests were determined by examining 400 sera from 4 herds known to be free of swine dysentery, and sera from immunised or experimentally infected pigs. Samples of serum from 40 bacon-weight pigs from each of the 106 herds were then collected at 2 abattoirs. Each serum was tested in enzyme-linked immunosorbent assays (ELISA) against the lipopolysaccharide of T hyodysenteriae of serogroups A, B and E, respectively. To assist in evaluating the test, 19 herds were resampled and retested, and faecal samples from 17 herds were cultured for T hyodysenteriae. Thirty-five of the 106 herds (33%) had serological evidence of infection when only one batch of sera from each herd was tested. The ELISA to detect T hyodysenteriae infection in herds using 40 sera was estimated as having a sensitivity of 77.3% and a specificity of 81.8% based on the owners' opinion of their herds disease status. Prevalence of infection within herds ranged from 2.5% to 47.5%, with a mean of 18%.     

Prevalence and spatial distribution of cattle herds infected with Theileria orientalis in New Zealand between 2012 and 2013

AIM: To describe the prevalence and spatial distribution of cattle herds infected with Ikeda and non-Ikeda types of Theileria orientalis in New Zealand between November 2012 and June 2013.

METHODS: Pooled serum samples collected historically between November 2012 and June 2013 were obtained from cattle herds throughout New Zealand. Each pooled sample consisted of approximately 20 individual cattle samples from that herd, and was provided with details of the spatial location of the herd (n=722). DNA from all samples was tested using two quantitative PCR assays for the detection of T. orientalis (all types) and the Ikeda type. The proportion of herds that were positive for T. orientalis and Ikeda type, or that were positive for T. orientalis but negative for Ikeda type (non-Ikeda positive) was determined for different regions of New Zealand.

RESULTS: The highest prevalence of herds infected with Ikeda type was detected in the Northland (33/35; 94%) and Auckland and the Waikato (63/191; 33%) regions. Only 2/204 (1%) herds were positive for the Ikeda type in the South Island. A high percentage of herds that were positive for non-Ikeda types was detected in the Gisborne and Hawkes Bay (23 (95%CI=13–37)%), Auckland and Waikato (22 (95%CI=16–29)%) and Bay of Plenty (24 (95%CI=10–44)%) regions.

CONCLUSIONS AND CLINICAL RELEVANCE: The high prevalence of Ikeda type detected in cattle herds in the Northland, Auckland and Waikato regions represents a risk to naive cattle being introduced into these regions. There is also the potential for resident cattle herds in the Gisborne and Hawkes Bay, Auckland, Waikato and Bay of Plenty regions to experience increased infection with the Ikeda type.

The overall impact experienced by regions will depend on other factors such as the number of herds present and the predominant type of farming, as well as the interplay between tick ecology, cattle immunity and movement patterns of cattle.