title: A study of bovine salivary proteins

authors: McIntosh, James Trevor

abstract: This physiological genetics approach to the study of the bovine salivary proteins was designed to: (a) Test the hypothesis that the salivary proteins, other than mucoprotein, might be related to, or involved in determining an animal’s susceptibility to bloat. (b) Gain some knowledge on the nature of the proteins in bovine saliva. This study paralleled the breeding programme selecting animals for high (HS) and low (LS) susceptibility to bloat. Two groups of adult cows, classified as HS or LS by field gradings (Group 1 and Group 2), and three groups of HS and LS progeny (1973, 1974 and 1975 animals), were studied.
Part I of this project involved studying saliva flow, total protein, and the eleven protein bands separated by polyacrylamide gel electrophoresis (PAGE) from saliva samples (collected from the mouth of cows), in relation to bloat grade or susceptibility.
Consistent results from these studies indicated that protein bands 4 and 6 were positively related to bloat susceptibility, and saliva flow and band 7 were negatively related to susceptibility. Band 7 was also negatively related to individual cow variance and positively related to saliva flow. It was demonstrated that a cow’s saliva flow could be markedly stimulated by the oral administration of acetic acid and this stimulation also increased the proportion and concentration of band 7.
The time since a cow last had access to feed which did not induce bloat, made very little difference to the parameters measured in saliva. However, in a series of Pasture Type Trials, where saliva samples were collected from animals directly after coming off grass or ‘bloat potent’ clover pasture, the level of band 4 was increased (shown as an environmental correlation between bloat grade and band 4). Band 6 showed a positive phenotypic correlation with bloat grade and a positive environmental correlation with saliva flow, while band 7 showed a positive correlation with protein.
Changes in bloat grade and the salivary parameters with age were followed in the 1973, 1974 and 1975 animals.
Results from studies in Part Two, indicated that immunoglobulins were present in saliva and a semi-purified protein fraction, band 1-2-3 complex, when run on SDS-electrophoresis was composed of subunits corresponding in molecular weight to those of IgG and secretory-IgA. Comparisons between saliva collected from the mandibular gland, mouth and oesophagus indicated that band 1 was secreted by the mandibular gland.
Band 6 was shown to contain two proteins one of which cross-reacted with anti-albumin serum, had the same electrophoretic mobility and molecular weight as serum albumin.
Band 7, 8 and 9 were statistically related and this relationship was demonstrated chemically by incubating saliva or semi-purified proteins, band 7-8-9 complex, with neuraminidase. The molecular weight of this complex was 65 300 (2 800 SEM) and appeared to be composed of three subunits with molecular weights of 26 000, 22 500 and 17 500.
Band 7 and 8 appeared to be secreted by the mandibular glands. 
Other studies included, screening for enzymic activity, SDS-electrophoretic studies to determine the range of proteins present, screening for cationic proteins, and electrophoretic and immunoelectrophoretic comparisons of mouth saliva with secretions from the mandibular, nasal, and nasolabial glands using whole saliva antiserum. 
It was concluded that protein bands 4, 6 and 7 and saliva flow were related to bloat susceptibility and that further work investigating the biological significance of these relationships was warranted.

text: A study of bovine salivary proteins This physiological genetics approach to the study of the bovine salivary proteins was designed to: (a) Test the hypothesis that the salivary proteins, other than mucoprotein, might be related to, or involved in determining an animal’s susceptibility to bloat. (b) Gain some knowledge on the nature of the proteins in bovine saliva. This study paralleled the breeding programme selecting animals for high (HS) and low (LS) susceptibility to bloat. Two groups of adult cows, classified as HS or LS by field gradings (Group 1 and Group 2), and three groups of HS and LS progeny (1973, 1974 and 1975 animals), were studied.
Part I of this project involved studying saliva flow, total protein, and the eleven protein bands separated by polyacrylamide gel electrophoresis (PAGE) from saliva samples (collected from the mouth of cows), in relation to bloat grade or susceptibility.
Consistent results from these studies indicated that protein bands 4 and 6 were positively related to bloat susceptibility, and saliva flow and band 7 were negatively related to susceptibility. Band 7 was also negatively related to individual cow variance and positively related to saliva flow. It was demonstrated that a cow’s saliva flow could be markedly stimulated by the oral administration of acetic acid and this stimulation also increased the proportion and concentration of band 7.
The time since a cow last had access to feed which did not induce bloat, made very little difference to the parameters measured in saliva. However, in a series of Pasture Type Trials, where saliva samples were collected from animals directly after coming off grass or ‘bloat potent’ clover pasture, the level of band 4 was increased (shown as an environmental correlation between bloat grade and band 4). Band 6 showed a positive phenotypic correlation with bloat grade and a positive environmental correlation with saliva flow, while band 7 showed a positive correlation with protein.
Changes in bloat grade and the salivary parameters with age were followed in the 1973, 1974 and 1975 animals.
Results from studies in Part Two, indicated that immunoglobulins were present in saliva and a semi-purified protein fraction, band 1-2-3 complex, when run on SDS-electrophoresis was composed of subunits corresponding in molecular weight to those of IgG and secretory-IgA. Comparisons between saliva collected from the mandibular gland, mouth and oesophagus indicated that band 1 was secreted by the mandibular gland.
Band 6 was shown to contain two proteins one of which cross-reacted with anti-albumin serum, had the same electrophoretic mobility and molecular weight as serum albumin.
Band 7, 8 and 9 were statistically related and this relationship was demonstrated chemically by incubating saliva or semi-purified proteins, band 7-8-9 complex, with neuraminidase. The molecular weight of this complex was 65 300 (2 800 SEM) and appeared to be composed of three subunits with molecular weights of 26 000, 22 500 and 17 500.
Band 7 and 8 appeared to be secreted by the mandibular glands. 
Other studies included, screening for enzymic activity, SDS-electrophoretic studies to determine the range of proteins present, screening for cationic proteins, and electrophoretic and immunoelectrophoretic comparisons of mouth saliva with secretions from the mandibular, nasal, and nasolabial glands using whole saliva antiserum. 
It was concluded that protein bands 4, 6 and 7 and saliva flow were related to bloat susceptibility and that further work investigating the biological significance of these relationships was warranted.

year: 1978

title: Dynamics of internal nutrient loading in a eutrophic, polymictic lake (Lake Rotorua, New Zealand)

authors: Burger, David F.

subjects: Nutrient pollution of water New Zealand Rotorua, Lake

abstract: ﻿Lake Rotorua is a large (80 krn2), polymictic, eutrophic, shalJow (mean depth 10.8 m) lake in central North Island, New Zealand. Blooms of cyanobacteria and occasional anoxia of bottom waters are characteristic of the water quality in the lake during summer months (Dec-Mar.). This study examines the dynamics of internal nutrient recycling processes in Lake Rotorua, including sediment nutrient release, sedimentation and resuspension, phytoplankton nutrient limitation and the relative importance of internal and external nutrient loads on influencing cyanobacteria biomass and summer bloom formation. 
Sediment nutrient release rates of phosphorus (P) in Lake Rotorua were estimated from changes in bottom water P concentration during a 19-day summer stratification event in February 2003. Changes in mass of hypolimnion P due to mixing events, together with settling, inflows, diffusion and regeneration, were factored into a simple model which was used to elucidate and quantify the complex nature of phosphorus fluxes in polymictic systems. Sediment soluble reactive phosphorus (SRP) release rates reached values of up to 26.3 mg m-2 d- 1 and increased during stratification, coinciding with a decline in bottom water dissolved oxygen (DO) concentrations. 
Phytoplankton nutrient limitation was examined in the lake using three in situ incubation experiments of 4 to 6 day duration in summer 2004. Two of the incubations were conducted during stratification, and one immediately after breakdown of stratification. Samples were enriched with either ammonium ( 1 mg NI-Lt L-1), phosphate (0.1 mg P04 L-1), or with both nutrients. A control with no nutrient addition was used for comparison. Phytoplankton responses to nutrient additions were determined at a species level from cell counts and at a community level from changes in chlorophyll a (chi-a) concentration. A simple phytoplankton growth model was applied to consider the interacting effects of P, nitrogen (N) and light limitation. Phytoplankton biomass generally responded to N plus P additions to a greater extent than with single nutrient additions alone, however, results were often not significant. Increase in community biomass was greater for P than N, and nutrient demand decreased after breakdown of stratification. Individual species responded differently to N and P additions, suggesting co-limitation, and that management of water quality in Lake Rotorua should restrict inputs of both N and P. Model results indicate that light also plays a major role in limiting phytoplankton biomass. 
Seasonal variations in sedimentation rates of total particulate matter (TPM), total phosphorus (TP), total nitrogen (TN) and chi-a were measured at three sites in the lake using cylindrical sediment traps. Deployment of traps at different depths at each site showed an increase in sedimentation rates of particulate inorganic material with trap depth, indicating that sediment resuspension is an important process in this lake. Resuspension was estimated to contribute up to 71 % of TPM sedimentation at the shallowest site. Net sedimentation rates across all sites, excluding resuspension fluxes, were 4.5 g m-2 d- 1 for TPM, and 19.8, 103.7 and 42.1 mg m-2 d-1 for TP, TN and chi-a, respectively. Sedimentation rates of all variables were highest in summer and at the deepest sampling site. Mean net sedimentation rates of N and P were between four and nine times greater, respectively, than estimates of net retention based on a nutrient mass balance, demonstrating that internal nutrient recycling is an important process in Lake Rotorua. 
Sediment release rates of SRP and Nlli were further determined seasonally at three sites (water depth 7, 14 and 20 m) in Lake Rotorua using in situ benthic chamber incubations. Rates of release of SRP ranged from 2.2 to 85.6 mg P m·2 d"1 and were largely independent of DO concentration. Two phases of Nlli release were observed in the chamber incubations; high initial rates of up to 2200 mg N m-2 d-1 in the first 12 h of deployment followed by lower rates of up to 270 mg N m-2 d-1 in the remaining 36 h of deployment. Releases of SRP and Nai were highest in summer and at the deepest of the three sites. High organic matter supply rates to the sediments may be important for sustaining high rates of sediment nutrient release. A nutrient budget of Lake Rotorua indicates that internal nutrient sources derived from benthic fluxes are more important than external nutrient sources to the lake. 
In the final part of this thesis, a vertically resolved water quality model, DYRESMCAEDYM, was used to assist with quantifying the relative contributions of internal and external nutrient inputs to the lake and their relative importance to cyanobacteria bloom formation. External nutrient loads were derived for 26 tributaries as well as for iii groundwater and stormwater flows. The total external load is 534 t yr-1 for TN and 34 t yr-1 for TP. Other forcing inputs to the model included meteorological data collected at a station beside the lake and discharge from the only outflow, Ohau Channel. Measured rates of sediment nutrient release obtained from benthic chamber measurements, profiles of water column nutrient concentrations, surface chl-a concentration and temperature and dissolved oxygen loggers were used to validate output from the DYRESM-CAEDYM model. Simulations of water column temperatures and SRP and NI-Li concentrations in Lake Rotorua showed a close representation of field measurements, and captured the timing and duration of stratification events and subsequent changes in bottom water nutrient concentrations. Model simulations of different nutrient loading scenarios indicate that reductions in sediment nutrient fluxes would be more effective in reducing cyanobacterial biomass than similar reductions in catchment fluxes, due to the incidence of large sediment nutrient release events in association with summer blooms. This finding indicates that only a significant and prolonged reduction in external loads, that in tum would reduce internal loads, will ultimately decrease cyanobacteria biomass in Lake Rotorua. 
In this study, field work and modelling were used to demonstrate the importance of internal nutrient loads in sustaining the current trophic status of Lake Rotorua. High external loading rates, coupled with high rates of organic matter sedimentation and sediment nutrient release rates, suggest that high phytoplankton biomass will remain a feature of this lake unless a significant and prolonged reduction in both N and P external loads is undertaken. The results of this study also emphasised the importance of conducting experimental measures of sedimentation and sediment nutrient release rates within eutrophic lakes, particularly where internal recycling processes may represent the dominant sources of nutrients to the lake.

text: Dynamics of internal nutrient loading in a eutrophic, polymictic lake (Lake Rotorua, New Zealand) ﻿Lake Rotorua is a large (80 krn2), polymictic, eutrophic, shalJow (mean depth 10.8 m) lake in central North Island, New Zealand. Blooms of cyanobacteria and occasional anoxia of bottom waters are characteristic of the water quality in the lake during summer months (Dec-Mar.). This study examines the dynamics of internal nutrient recycling processes in Lake Rotorua, including sediment nutrient release, sedimentation and resuspension, phytoplankton nutrient limitation and the relative importance of internal and external nutrient loads on influencing cyanobacteria biomass and summer bloom formation. 
Sediment nutrient release rates of phosphorus (P) in Lake Rotorua were estimated from changes in bottom water P concentration during a 19-day summer stratification event in February 2003. Changes in mass of hypolimnion P due to mixing events, together with settling, inflows, diffusion and regeneration, were factored into a simple model which was used to elucidate and quantify the complex nature of phosphorus fluxes in polymictic systems. Sediment soluble reactive phosphorus (SRP) release rates reached values of up to 26.3 mg m-2 d- 1 and increased during stratification, coinciding with a decline in bottom water dissolved oxygen (DO) concentrations. 
Phytoplankton nutrient limitation was examined in the lake using three in situ incubation experiments of 4 to 6 day duration in summer 2004. Two of the incubations were conducted during stratification, and one immediately after breakdown of stratification. Samples were enriched with either ammonium ( 1 mg NI-Lt L-1), phosphate (0.1 mg P04 L-1), or with both nutrients. A control with no nutrient addition was used for comparison. Phytoplankton responses to nutrient additions were determined at a species level from cell counts and at a community level from changes in chlorophyll a (chi-a) concentration. A simple phytoplankton growth model was applied to consider the interacting effects of P, nitrogen (N) and light limitation. Phytoplankton biomass generally responded to N plus P additions to a greater extent than with single nutrient additions alone, however, results were often not significant. Increase in community biomass was greater for P than N, and nutrient demand decreased after breakdown of stratification. Individual species responded differently to N and P additions, suggesting co-limitation, and that management of water quality in Lake Rotorua should restrict inputs of both N and P. Model results indicate that light also plays a major role in limiting phytoplankton biomass. 
Seasonal variations in sedimentation rates of total particulate matter (TPM), total phosphorus (TP), total nitrogen (TN) and chi-a were measured at three sites in the lake using cylindrical sediment traps. Deployment of traps at different depths at each site showed an increase in sedimentation rates of particulate inorganic material with trap depth, indicating that sediment resuspension is an important process in this lake. Resuspension was estimated to contribute up to 71 % of TPM sedimentation at the shallowest site. Net sedimentation rates across all sites, excluding resuspension fluxes, were 4.5 g m-2 d- 1 for TPM, and 19.8, 103.7 and 42.1 mg m-2 d-1 for TP, TN and chi-a, respectively. Sedimentation rates of all variables were highest in summer and at the deepest sampling site. Mean net sedimentation rates of N and P were between four and nine times greater, respectively, than estimates of net retention based on a nutrient mass balance, demonstrating that internal nutrient recycling is an important process in Lake Rotorua. 
Sediment release rates of SRP and Nlli were further determined seasonally at three sites (water depth 7, 14 and 20 m) in Lake Rotorua using in situ benthic chamber incubations. Rates of release of SRP ranged from 2.2 to 85.6 mg P m·2 d"1 and were largely independent of DO concentration. Two phases of Nlli release were observed in the chamber incubations; high initial rates of up to 2200 mg N m-2 d-1 in the first 12 h of deployment followed by lower rates of up to 270 mg N m-2 d-1 in the remaining 36 h of deployment. Releases of SRP and Nai were highest in summer and at the deepest of the three sites. High organic matter supply rates to the sediments may be important for sustaining high rates of sediment nutrient release. A nutrient budget of Lake Rotorua indicates that internal nutrient sources derived from benthic fluxes are more important than external nutrient sources to the lake. 
In the final part of this thesis, a vertically resolved water quality model, DYRESMCAEDYM, was used to assist with quantifying the relative contributions of internal and external nutrient inputs to the lake and their relative importance to cyanobacteria bloom formation. External nutrient loads were derived for 26 tributaries as well as for iii groundwater and stormwater flows. The total external load is 534 t yr-1 for TN and 34 t yr-1 for TP. Other forcing inputs to the model included meteorological data collected at a station beside the lake and discharge from the only outflow, Ohau Channel. Measured rates of sediment nutrient release obtained from benthic chamber measurements, profiles of water column nutrient concentrations, surface chl-a concentration and temperature and dissolved oxygen loggers were used to validate output from the DYRESM-CAEDYM model. Simulations of water column temperatures and SRP and NI-Li concentrations in Lake Rotorua showed a close representation of field measurements, and captured the timing and duration of stratification events and subsequent changes in bottom water nutrient concentrations. Model simulations of different nutrient loading scenarios indicate that reductions in sediment nutrient fluxes would be more effective in reducing cyanobacterial biomass than similar reductions in catchment fluxes, due to the incidence of large sediment nutrient release events in association with summer blooms. This finding indicates that only a significant and prolonged reduction in external loads, that in tum would reduce internal loads, will ultimately decrease cyanobacteria biomass in Lake Rotorua. 
In this study, field work and modelling were used to demonstrate the importance of internal nutrient loads in sustaining the current trophic status of Lake Rotorua. High external loading rates, coupled with high rates of organic matter sedimentation and sediment nutrient release rates, suggest that high phytoplankton biomass will remain a feature of this lake unless a significant and prolonged reduction in both N and P external loads is undertaken. The results of this study also emphasised the importance of conducting experimental measures of sedimentation and sediment nutrient release rates within eutrophic lakes, particularly where internal recycling processes may represent the dominant sources of nutrients to the lake.

year: 2006

title: Ngā Pakanga mō Wāhi Tapu: Battles over sacred places

authors: Joseph, Robert

abstract: Wāhi tapu are the sacred places of Māori in Aotearoa New Zealand. All cultures have sacred sites such as cemeteries, significant battlefields, and places of spiritual significance. Standout examples internationally include the ‘Weeping Wall’ in the Holy Land, Stonehenge in Britain, Maraeātea in the Pacific, and Intihuatana at Macchu Picchu in Peru. But unlike these sites, wāhi tapu are rarely a visible structure but rather a site or area within the landscape with values so significant that restrictions are warranted. In law, wāhi tapu are defined as a ‘place of special significance according to tikanga Māori’
 and ‘a place sacred to Māori in the traditional, spiritual, religious, ritual, or mythological sense’.
As the aphorism above emphasises, Māori historically were willing to die for land (and for women of course!), and wāhi tapu are the most sacred of places. Many battles have been fought over actual or proposed transgressions of their sacredness. In earlier years, blood was shed and men did die; today battles tend to occur in the Environment Court, but are just as passionate. Recent decades have seen conflicts where wāhi tapu were threatened by new highways, railroads, airports, prisons, residential and commercial developments, wind farms, power stations and waste dumps.
In this chapter, I discuss a number of the battles that have been fought over the development or preservation of wāhi tapu from first European contact up until the present time. I highlight the growing legal recognition of tikanga Māori in relation to these sacred landscapes, and conclude with a proposed pragmatic solution for all New Zealanders when negotiating developments over wāhi tapu. Having a
background in law and tribal affiliations, I discuss these tensions from both legal and kaupapa Māori perspectives.

text: Ngā Pakanga mō Wāhi Tapu: Battles over sacred places Wāhi tapu are the sacred places of Māori in Aotearoa New Zealand. All cultures have sacred sites such as cemeteries, significant battlefields, and places of spiritual significance. Standout examples internationally include the ‘Weeping Wall’ in the Holy Land, Stonehenge in Britain, Maraeātea in the Pacific, and Intihuatana at Macchu Picchu in Peru. But unlike these sites, wāhi tapu are rarely a visible structure but rather a site or area within the landscape with values so significant that restrictions are warranted. In law, wāhi tapu are defined as a ‘place of special significance according to tikanga Māori’
 and ‘a place sacred to Māori in the traditional, spiritual, religious, ritual, or mythological sense’.
As the aphorism above emphasises, Māori historically were willing to die for land (and for women of course!), and wāhi tapu are the most sacred of places. Many battles have been fought over actual or proposed transgressions of their sacredness. In earlier years, blood was shed and men did die; today battles tend to occur in the Environment Court, but are just as passionate. Recent decades have seen conflicts where wāhi tapu were threatened by new highways, railroads, airports, prisons, residential and commercial developments, wind farms, power stations and waste dumps.
In this chapter, I discuss a number of the battles that have been fought over the development or preservation of wāhi tapu from first European contact up until the present time. I highlight the growing legal recognition of tikanga Māori in relation to these sacred landscapes, and conclude with a proposed pragmatic solution for all New Zealanders when negotiating developments over wāhi tapu. Having a
background in law and tribal affiliations, I discuss these tensions from both legal and kaupapa Māori perspectives.

year: 2011