Purification of secretory immunoglobulin A from milk of the brushtail possum (Trichosurus vulpecula)

AIM: To identify and purify secretory immunoglobulin A (sIgA), a key effecter molecule in mucosal immune responses, from milk of the brushtail possum (Trichosurus vulpecula).

METHODS: Milk samples were collected from female possums with pouch young, and clarified by centrifugation and precipitation methods. The clarified fraction was purified by gel filtration and affinity chromatography to yield sIgA. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting techniques were used to assess the purity of the final product, and to identify the heavy (H) chain, light (L) chain and secretory component (SC) of possum sIgA.

RESULTS: Immunoblotting, using antibodies raised against cloned possum sIgA SC and H-chain, and a synthetic peptide fragment of the H-chain, confirmed the identity of the purified protein. The N-terminal amino acid sequence of purified possum sIgA showed strong homology to reported sequences of H-chain variable regions of marsupial immunoglobulins.

CONCLUSIONS: Milk was shown to be a convenient source of mucosal secretion containing sIgA, and a process involving 2 precipitation and 2 chromatography steps produced purified sIgA. This IgA preparation will prove useful for the generation of sIgA-specific immunological reagents for measurement of immune responses in the development of mucosal-based vaccines for biological control of possums.     

Improving the Reproductive Efficiency by Zoo-Technical Methods at a Dairy Farm

This trial was conducted on a Hungarian dairy farm between July 2001 and December 2004. The objective of this work was to improve the reproductive efficiency with relatively high milk production. At the beginning of this trial blood and fodder samples were taken for checking the metabolic status of the animals in order to determine their health condition. The nutritive value of the daily ration for all groups met with the requirements of the Hungarian National Standard and almost all serum metabolic parameters differed between the milking and pregnant animals. Early pregnancy detection (by ultrasound) and ovulation synchronizing methods were introduced to optimize reproductive performance. The oestrus cycle was also checked by ultrasound and open cows were treated by the appropriate method in order to inseminate them as soon as possible. Efficiency of artificial insemination (AI) followed by a single prostaglandin F2alpha (PGF2alpha) and Ovsynch treatment was similar (30.8% and 29%) and less effective than AI after natural heat detection (37.1%). Provsynch (Pre-synch followed by Ovsynch) was the most effective ovulation synchronization method (conception rate=42.6%; p<0.01). Although milk production increased between 2002 and 2004 by approximately 600 kg per cow, the calving period decreased by 20 days and the number of AIs per pregnancy was also improved (0.8 AI per conception). These findings are really beneficial economically because the decrease in the calving interval returns profit for the dairy farms; one open day costs euro2.5/cow. Economical analysis showed a high profit ratio from the reproductive 'investment' on this farm. Every invested euro yielded approximately euro10.     

Radioactive gold-198 seeds for the treatment of squamous cell carcinoma in the eyelid of a cat

A 19-year-old, speyed, domestic short-hair cat was presented with an eyelid tumour of 3 months duration. Ophthalmic examination revealed a large, raised, ulcerated and bleeding mass affecting the left lower eyelid. The mass was 12 mm x 10 mm in size, extended to the medial canthus and had eroded 10 mm of eyelid margin. Mandibular lymph nodes were not palpably enlarged. A diagnosis of squamous cell carcinoma was confirmed by histologic examination. Due to involvement of more than half the lower eyelid, it was decided that treatment with radioactive gold-198 seeds was appropriate. Nine radioactive gold-198 seeds were implanted, delivering a minimum tumour dose of 65 Gy. Three weeks postoperatively the mass had resolved, with only a small scab remaining. After a further 3 weeks, there was no evidence of the eyelid tumour. Although approximately 7 mm of eyelid margin was absent, this did not cause any clinical signs other than mild conjunctivitis. There was no evidence of the tumour 10 months postoperatively, when the cat was euthanased for unrelated disease.     

Listeria monocytogenes gastroenteritis in sheep

Extract

Listeria monocytogenes is widely distributed in the environment of farm animals. It is found in soil, faeces and nasal secretions of healthy animals, water troughs and animal feeds. Under certain conditions it becomes pathogenic, causing serious disease in cattle and sheep. Most commonly it causes meningoencephalitis but, on occasion, septicaemic listeriosis results in abortion, and more recently it has been associated with gastroenteritis.     

Presence of Leptin and Its Receptor in the Hypothalamus,Uterus and Ovaries of Swine Females Culled with Distinct Ovarian Statuses and Parities

Leptin acts on energy metabolism, affecting reproductive functions through activation of its receptors in the brain and in reproductive organs. This study compared the presence of leptin and its receptor (ObR‐b) in hypothalamus neurons, endometrial glands and oocytes of culled swine females across ovarian statuses and parities. Immunohistochemistry was done in samples of uterus, ovaries and hypothalamus from 28 culled females, using polyclonal antibodies antileptin and ObR‐b. Immunolabelling was compared for sows categorized by parity at culling (0, 1, 2–4 and <4) and ovarian status (luteal and follicular phases of the oestrous cycle and with cysts). Immunolabelling for leptin and ObR‐b in neurons and oocytes was weaker in females with cysts (p < 0.05) than in those at the follicular phase. In endometrial glands, leptin immunolabelling was less intense in females with cysts (p < 0.05), but immunolabelling for ObR‐b was similar across ovarian statuses (p > 0.05). In sows culled with 2–4 parities, leptin immunolabelling in neurons and endometrial glands was more intense than in nulliparous females (p < 0.05). In comparison with sows culled at greater parities, ObR‐b immunolabelling for nulliparous females was less intense in endometrial glands and in oocytes (p < 0.05), but more intense in neurons (p < 0.05). Thus, in swine, the presence of leptin and ObR‐b varies across parities and is more intense in the uterus, ovaries and hypothalamus of females that were cycling before culling than in those having cystic ovaries.     

Intra-uterine transmission of Mycobacterium avium subsp paratuberculosis in subclinically affected red deer (Cervus elaphus)

AIM: To determine the rate of transmission of Mycobacterium avium subsp paratuberculosis (M. ptb) from hind to fetus in utero, and the risk of transmission from dam to fawn via infected colostrum and milk in subclinically affected red deer hinds.

METHODS: Hinds were sourced from farms in Otago or Southland and selected for the study if they were positive to the immunoglobulin G1 (IgG1) modified enzyme-linked immunosorbent assay (ELISA) (Paralisa) and exhibited no clinical signs of Johne's disease. The hinds (n=35) were sent to a deer slaughter premises (DSP; n=31) or were killed on-farm (n=4). All post-mortem samples were collected from the fetus first and then from the dam, taking care to avoid cross contamination between samples. Fresh samples (n=185) were collected for culture, and tissue samples (n=72) were collected from 24 hinds and their fetuses for histopathological examination.

RESULTS: A total of 24/35 hinds selected were suitable for inclusion in the study. Eighteen of these pregnant hinds were culture-positive for M. ptb, and 14 of these had culture-positive fetuses, representing a transmission rate of 78% (95% confidence interval (CI) =0.58–0.98) from dam to fetus. Of the 16 mammary glands sampled, 11 (69%) were culture-positive for M. ptb while 12/15 (80%) mammary lymph nodes sampled were also culture-positive.

CONCLUSIONS: This study demonstrated a high rate of transmission of M. ptb from dam to fetus in red deer, and a potential risk of transmission to fawns suckling from mothers that are subclinically affected with Johne's disease.     

Field evaluation of a mixture of albendazole sulphoxide and levamisole against Ostertagia and Trichostrongylus spp in sheep

The efficacy of a mixture of albendazole sulphoxide and levamisole, 3.6 and 8.25 mg/kg respectively, at single and double dose rates, was compared with the recommended dose rate of each anthelmintic alone. The comparison was conducted on groups of 6 to 14-week-old lambs on 22 farms, 16 of which had evidence of multiple resistance to benzimidazole and levamisole. A single dose of the mixture reduced mean egg counts by 95% on half the farms with multiple resistance and on all the remaining farms. Consequently, the mixture should be included in an assessment of effective anthelmintics on farms to determine its value for nematode control. A double dose rate of mixture was effective on all but 4 farms. Reductions caused by the mixture were due to the additive actions of the drugs on 18 of 22 farms; synergistic action was noted on only 3 farms. It was concluded that the mixture of albendazole sulphoxide and levamisole offered many farmers an effective anthelmintic for use in preventive control programs. Recommendations for such programs include annual rotation of effective anthelmintics as a means of delaying selection for drug resistance.