The effect of cultivated mixed-species green fodder on intake, milk production and milk composition of housed dairy goats.

The majority of New Zealand dairy goat farmers utilise cultivated green-fed fodder dominated by perennial ryegrass (Lolium perenne L.) and white clover (Trifolium repens L.), but evidence from other ruminant species suggests that milk production may be improved when using a more diverse array of species within the green fodder. The aim of this experiment was to determine whether feeding lactating dairy goats a mixed-species green fodder (MF, consisting of perennial ryegrass, timothy (Phleum pratense L.), prairie grass (Bromus willdenowii Kunth), white clover, red clover (Trifolium pratense L.), lucerne (Medicago sativa L.), chicory (Cichorium intybus L.) and plantain (Plantago lanceolata L.) improves dietary intake, milk yield and composition compared with a standard ryegrass and white clover green fodder (SF). Thirty-six mid-lactation goats were housed indoors in pairs and split into two groups (A and B). The trial was split into three periods - firstly a uniformity period of 6 days, in which all goats were fed a combination of both green fodder types, followed by two treatment periods (P1 and P2) of 12 days, respectively. For P1, group A was fed MF and group B was fed SF, and then the group diets were switched for P2. Goats fed MF had 13% greater dry matter intake and 7% greater milk yield than goats fed SF. In addition, the milk protein and fat concentration of goats fed MF were 4% greater than for those fed SF, whereas there was no effect on milk lactose concentration. There was no treatment effect on the levels of protein, glucose, urea or non-esterified fatty acids in the blood of the goats. An effect of green fodder type on milk fat profile was demonstrated, with proportions of pentadecylic acid (C15:0), cis-vaccenic acid (C18:1 c11), linoleic acid (C18:2 n6) and α-linolenic acid (C18:3 n3) being increased in response to MF consumption. In contrast, iso-C15 and iso-C17 proportions were lesser. In summary, this study demonstrated that goats fed MF increased green fodder intake and milk production compared with goats fed SF. The green fodder type affected the fatty acid profile of goat's milk, with MF increasing the levels of beneficial polyunsaturated omega fatty acids (linoleic and α-linolenic acids).

Cell survival signaling in the bovine mammary gland during the transition from lactation to involution.

In dairy cows, mammary gland involution, and thus a decline in milk production, occurs following peak lactation. To examine the cell signaling pathways regulating involution of the mammary gland, signal transducer and activator of transcription factors (STAT5 and 3), suppressors of cytokine signaling (SOCS1-3 and CIS), insulin-like growth factors (IGF1 and 2), and protein kinase B (Akt) were examined. Mammary involution was induced by termination of milking, and alveolar tissue was collected from 52 nonpregnant, primiparous, mid-lactation Holstein-Friesian cows killed at 0, 6, 12, 18, 24, 36, 72, and 192h postmilking. Qualitative immunohistochemistry showed that activated (phosphorylated) STAT5-P was localized in nuclei of mammary epithelial cells at the early time points, with detection levels decreasing by 24h postmilking. In contrast, STAT3-P was barely detectable at the early time points, with detection levels increasing following longer postmilking periods. This was supported by Western analysis, which showed a decline in STAT5 and STAT5-P protein levels by 24h postmilking, no change in STAT3 levels, and an increase in STAT3-P protein (barely detectable at the early time points) by 72h postmilking. Quantitative real-time reverse transcription PCR analysis showed SOCS1 and SOCS3 mRNA increased by 72h postmilking compared with 6h postmilking. The SOCS2 mRNA remained unchanged across the time series, whereas CIS decreased by 18h postmilking and remained lower compared with that at 6h postmilking until 72h postmilking. The IGF1 mRNA increased by 192h postmilking, whereas IGF2 mRNA decreased by 18h postmilking compared with 6h postmilking. The IGFBP5 mRNA and protein levels of Akt and Akt-P remained unchanged over the time series. These results show that reciprocal activation of STAT5 and STAT3 occurs at the onset of mammary gland involution in the bovine, albeit at a slower rate than in rodents. Mathematical modeling of the pathways indicated that activated STAT3 could block the STAT5 pathway by upregulating SOCS3. The regulation of IGF1-Akt signaling suggests that by 192h postmilking in dairy cows, the involution process is still in the reversible phase, with quiescent mammary epithelial cells not yet in the senescent phase.

The effects of milking frequency on insulin-like growth factor I signaling within the mammary gland of dairy cows.

In dairy cows, short-term changes in milking frequency (MF) in early lactation have been shown to produce both an immediate and a long-term effect on milk yield. The effect of MF on milk yield is controlled locally within mammary glands and could be a function of changes in either number or activity of secretory mammary epithelial cells (MEC). Insulin-like growth factor I (IGF-I) signaling is one candidate factor that could mediate these effects, as it can be controlled locally within mammary glands. Both MEC number and activity can be affected by IGF-I signaling by activating the phosphoinositide 3-kinase (PI3K)/Akt and extracellular-signal-regulated kinase (ERK)1/2 pathways. To investigate the relationship between MF and IGF-I signaling, udder halves of 17 dairy cows were milked either 4 times a day (4×) or once a day (1×) for 14 d in early lactation. On d 14, between 3 and 5 h following milking, mammary biopsies were obtained from 10 cows from both udder halves, and changes in the expression of genes associated with IGF-I signaling and the activation of the PI3K/Akt and ERK1/2 pathways were measured. The mRNA abundance of IGF type I receptor, IGF binding protein (IGFBP)-3, and IGFBP-5 were lower following 4× milking relative to 1× milking. However, the mRNA abundance of IGF-I was not affected by MF. Both IGFBP3 and IGFBP5 are thought to inhibit IGF-I; therefore, decreases in their mRNA abundance may serve to stimulate the IGF-I signal in the 4×-milked mammary gland. The activation of PI3K/Akt pathway was lower in response to 4× milking relative to 1×, and the activation of the ERK1/2 was unaffected by MF, suggesting that they do not mediate the effects of MF.

Activation of signal transducer and activator of transcription 5 (STAT5) is linked to ß1-integrin protein abundance in unilaterally milked bovine mammary glands.

Prolactin (PRL) is important in the regulation of milk synthesis in mammary epithelial cells (MEC). In cattle, circulating levels of PRL are not limiting, suggesting the possible involvement of other factors that may control the response to PRL at the cellular level. The effects of milking frequency (MF) on milk synthesis are controlled locally within mammary glands and involve PRL signaling. To further investigate this relationship between MF and PRL signaling, udder halves of 17 dairy cows were milked either 4 times a day (4×) or once a day (1×) for 14 d in early lactation. Mammary biopsies were obtained 3 to 5h following milking from both udder halves of 10 cows, and changes in PRL and associated pathways were measured. The abundance of STAT5A mRNA was higher after 4× milking, whereas that of the PRL receptor (PRLR) and STAT3 were lower relative to that after 1× milking. In 4× mammary tissues, the protein levels of STAT5, activated STAT5, and ß1-integrin were higher, whereas the those of the long isoform of PRL receptor and activated STAT3 were lower than 1× tissues. The activation of STAT5 correlated strongly with major milk protein mRNA abundance (r=0.86 to 0.94) and ß1-integrin protein levels (r=0.91). These results confirm that major milk protein gene expression is associated with STAT5 activation and suggests that the STAT5 and ß1-integrin signaling pathways are linked. Modulation of ß1-integrin abundance in response to changes in MF may be a mechanism that controls the MEC ability to respond to PRL and therefore its secretory activity.

The effects of milking frequency in early lactation on milk yield, mammary cell turnover, and secretory activity in grazing dairy cows.

In dairy cows, short-term changes of milking frequency in early lactation have been shown to produce an immediate and a long-term effect on milk yield in stall-fed cows. The effect is controlled locally within mammary glands and could be a function of either secretory mammary epithelial cell number or activity. To resolve this and determine its applicability in other feed management systems, a unilateral milking frequency experiment was conducted with udder halves of 17 multiparous, pasture-fed dairy cows milked either 4 times (4×) or once a day (1×) for 14d from 5±2d in milk. Mean half-udder milk yield during the treatment period was higher from the 4× compared with 1× udder halves and continued to be higher until 200d in milk once returned to twice a day milking. Mammary biopsies were obtained on d 14 of treatment from both udder halves of 10 cows. Proliferation of mammary cells was higher in 4× udder halves compared with 1×, whereas no difference in apoptosis levels was detected. Abundance of αS1-casein, ß-casein, α-lactalbumin, and ß-lactoglobulin mRNA was higher in tissue samples from 4× udder halves compared with 1×, whereas lactoferrin mRNA abundance was lower in 4× udder halves. In summary, change in milking frequency during early lactation affects proliferation of mammary cells as well as expression of the major milk protein genes, which both contribute to the observed changes in milk yield during and after unilateral milking frequency treatment.

Newly attenuated Mycobacterium bovis mutants as vaccines against bovine tuberculosis, particularly for possums.

Bovine tuberculosis costs New Zealand more than $80 million per year, mostly because extensive areas of the country are occupied by brushtail possums infected with Mycobacterium bovis. AgResearch has a major programme to produce new live tuberculosis vaccines that can be delivered to possums. Primary work involved development of molecular biological methods to enable genetic manipulation of M. bovis, including the production of random and specific mutants. Many avirulent mutants of M. bovis have been produced and their vaccine efficacy has been compared to BCG in guinea pigs. Selected mutants that perform at least as well as BCG are retested in guinea pigs using an extended vaccination protocol in which animals are pre-sensitized to environmental mycobacteria to mimic natural exposure. Ten candidate vaccines that have induced good protection in guinea pigs have been subsequently tested as vaccines in possums. While the protective efficacy of an M. bovis mutant inoculated into guinea pigs reliably indicated that some protection would be induced in possums, the most protective mutant in guinea pigs was different from that in possums. This illustrates the importance of testing in the target species as part of new vaccine development. An important outcome of this work was the identification of an operon in M. bovis whose inactivation produced an avirulent M. bovis vaccine candidate that was better than BCG in protecting possums from experimental tuberculosis. Allelic exchange methods are now being used to produce vaccine strains with multiple specific mutations to improve safety and immunological characteristics.

Virus-like particles in the ovaries of Microctonus aethiopoides Loan (Hymenoptera: Braconidae): comparison of biotypes from Morocco and Europe.

Virus-like particles (MaVLP) have been discovered in the ovarial epithelial cells of the solitary, koinobiont, endoparasitoid, Microctonus aethiopoides Loan (Hymenoptera: Braconidae) introduced to New Zealand originally from Morocco to control the lucerne pest Sitona discoideus Gyllenhal (Coleoptera: Curculionidae). MaVLP have been found in all females examined. It has been suggested, although not demonstrated, that like many other such VLP found in parasitoids, MaVLP might play a role in host immunosuppression. Since another biotype of M. aethiopoides from Ireland has been proposed for introduction to control the white clover pest, Sitona lepidus Gyllenhal, in New Zealand, it was considered that females from this biotype warranted transmission electron microscope examination for VLP. No VLP were observed in ovarian tissues of specimens collected from three different locations in Ireland. Similarly, none were found in M. aethiopoides sourced from France, Wales, and Norway. These observations are discussed in relation to quarantine host specificity tests with the Irish biotype, which found that the host range of the Irish biotype is likely to be less extensive than that of the Moroccan biotype already in New Zealand.

High-amplitude peristaltic contractions in a patient with esophageal intramural pseudodiverticulosis.

In their initial radiographic description of esophageal intramural pseudodiverticulosis (EIP) in 1960, Mendl and coworkers suggested that elevated esophageal intraluminal pressure might be etiologically important. Chronic inflammation and moniliasis have also been implicated. A patient is reported with EIP and a primary esophageal motility disorder characterized by esophageal contractions of increased amplitude and duration. Features confusing the interpretation of esophageal motor abnormalities in previously reported cases, such as diabetes mellitus, fungal esophagitis, and stricture formation, were not present in this case.