Uncertainty and urban water recharge for managing groundwater availability using decision support.

Quantifying groundwater availability depends upon sound methods and the use of integrated models. To determine availability or sustainable yield, the influence of scientific uncertainty from key sources, such as anthropogenic recharge, must be considered. This study evaluates uncertainty in recharge interpretations on the modeled available water balance for an urban case in Texas, USA. Analyses are completed using the Groundwater Decision Support System, which is a research code-base for an integrated modeling. The case study develops spatially and temporally resolved recharge interpretations based on NEXRAD precipitation and detailed land use data. Results demonstrate the implications of scientific uncertainty as it influences recommendations for policy and urban water management decisions that are based on modeled outputs. Geospatial methods account for spatial and temporal components and can be replicated for other systems. These methods are also useful for resolving uncertainty in relation to the influence of urbanization on recharge through land use change.

PCR examination of bronchoalveolar lavage samples is a useful tool in pre-clinical diagnosis of ovine pulmonary adenocarcinoma (Jaagsiekte).

Ovine pulmonary adenocarcinoma (OPA) is a contagious lung tumour of sheep caused by Jaagsiekte sheep retrovirus (JSRV). The disease is a particular problem in flocks in many parts of the world. The aim of the study was to assess screening methods for individual animals as a prelude to future eradication trials. Results of histological examination were used as the standard to evaluate the relative sensitivity and specificity of an established heminested polymerase chain reaction (PCR) test for JSRV proviral DNA from blood and bronchoalveolar lavage (BAL) samples. PCR results from tissue samples are included as control data. PCR testing of blood samples was found to have an estimated sensitivity of only 10% (95% confidence interval (CI) 3-20) while the sensitivity of the PCR test on BAL samples was 89% (CI 79-96) in comparison to the results of histological examination. We conclude that PCR testing of BAL samples is an effective confirmatory test for sheep with suspected clinical OPA. It is also a useful tool for the pre-clinical identification of individual infected sheep within an infected flock and therefore may prove beneficial in future control or eradication programmes.

Eradication of ovine pulmonary adenocarcinoma by motherless rearing of lambs.

The principles of maedi-visna eradication programmes were applied to a field trial for the eradication of ovine pulmonary adenocarcinoma (OPA). In two maternal flocks the prevalence of gross and histological lesions in slaughtered animals was 18.3 per cent and 29.8 per cent, respectively. The lambing period was supervised for three consecutive years from 1999 to 2001, during which the lambs were taken away from their mothers at birth, deprived of maternal colostrum, and hand-reared away from other sheep. Over the three-year period, 322 hand-reared animals, mainly male lambs between 10 and 14 months old, were slaughtered; their lungs were examined grossly, 52.5 per cent of them were examined histologically, and 105 samples of caudal mediastinal lymph nodes were examined by PCR. No OPA tumours were detected in the slaughter specimens from the derived flock, but one lamb had histological lesions in one lung location; intrauterine transmission was ruled out in this case. No clinical OPA has subsequently been observed in the hand-reared flock. Bronchoalveolar lavage samples from the breeding stock were examined by PCR in order to rule out further subclinical cases of OPA. No Jaagsiekte retrovirus was detected in any of the 488 samples.

The molecular epidemiology of Mycobacterium bovis infections in Tanzania.

A molecular epidemiological study to determine the zoonotic importance of bovine tuberculosis was carried out in Tanzania. Specimens from human cases of tuberculosis as well as from slaughtered cattle were collected from regions with a high proportion of extrapulmonary tuberculosis. In order to determine the similarity of strains from the two sources, molecular typing techniques, namely RFLP and spoligotyping, were used to determine the genetic profile of the strains involved. The results of pTBN12 typing of M. bovis from cattle and man has shown a rather heterogeneous population of this species spread all over Tanzania, assuming that the present sample is representative. There were 13 different pTBN12 RFLP types encountered. The genetic relatedness between the pTBN12 RFLP patterns indicated a high degree of relatedness (86%) between the dominant pTBN12 genotypes existing in Tanzania. There were 13 different spoligotypes found in this study, whose genetic relatedness was also high (79%). DNA profiles were also confirmed by IS986 RFLP, which revealed that strains have 1-13 copies of IS986. Geographically, there was overlap between pTBN12 RFLP and spoligotypes amongst strains isolated from various parts of Tanzania. The diversity of the RFLP and spoligotype patterns observed in Tanzania probably reflects the extensive internal movements of cattle belonging to pastoralists. The evidence of overlap between DNA fingerprints of M. bovis from cattle and man has once more highlighted a need for synergy of veterinary and medical policies in the control of tuberculosis in Tanzania and probably in other developing countries.

In-situ demonstration of mitogen-activated protein kinase Erk 1/2 signalling pathway in contagious respiratory tumours of sheep and goats.

Ovine pulmonary adenocarcinoma (OPA) and enzootic nasal adenocarcinoma (ENA) are two contagious neoplastic diseases of secretory epithelial cells in the respiratory system of sheep and goats. Jaagsiekte sheep retrovirus (JSRV) is the aetiological agent of OPA, and enzootic nasal tumour virus (ENTV) is associated with ENA. The genomes of these retroviruses do not contain known oncogenes but products of the env gene are important in the generation of transforming stimuli. However, the cell signalling pathways activated in vivo are not completely understood. This study was based on the use of activation stage antibodies specifically detecting proteins of the extracellular signal regulated kinase Erk 1/2 cell signalling pathway and transcription factors. Tissue sections were collected from four natural cases of OPA, four experimentally induced OPA tumours, four ENA tumours in sheep, four ENA tumours in goats, two normal sheep lungs and two lungs with chronic inflammation. Routine immunohistochemical procedures with phosphorylation stage-specific antibodies were carried out. Representative proteins of the Erk1/2 pathway (Raf-1, Mek1/2 and p44/42MAPK) were activated in natural cases of OPA and ENA in sheep and goats and also in experimentally induced OPA. Transcription factors 90Rsk and Elk-1 were activated in OPA and ENA tumours. However, c-Myc was activated only in OPA tumours. In contagious respiratory neoplasms of sheep and goats the Erk1/2 pathway appears to be important for the in-vivo generation of the transforming stimuli.

Experimental coinduction of type D retrovirus-associated pulmonary carcinoma and lentivirus-associated lymphoid interstitial pneumonia in lambs.

Two retrovirus-associated pulmonary diseases of sheep [ovine pulmonary carcinoma (OPC); sheep pulmonary adenomatosis], a bronchoalveolar carcinoma, and lymphoid interstitial pneumonia (LIP) were induced simultaneously in 9 of 9 neonatal lambs. The lambs were killed 8-28 weeks after intratracheal injection of lung tumor homogenate or lung fluid derived from sheep with naturally occurring OPC and ovine lentivirus (OvLV) infection. The inoculated lambs developed multifocal neoplasms of alveolar type II cells or nonciliated bronchiolar epithelial cells, LIP, and pulmonary lymph node hyperplasia, and all produced antibody to OvLV. OvLV was isolated from 6 to 7 lambs tested, and infectious center assay of pulmonary lavage cells from 3 lambs revealed that approximately 1 in 1,000 pulmonary lavage cells contained infectious lentivirus. Neither contact control lambs nor control lambs that received ultrafiltered lung fluid developed evidence of either disease or of OvLV infection. Lung fluid or tumor tissue of lambs with OPC contained a 26,000-dalton protein that cross-reacted with antiserum to p27 to Mason-Pfizer monkey virus, a type D retrovirus. The fact that no antigenic cross-reaction between OvLV and type D retroviruses has been demonstrated supports the presence of two retroviruses in sheep with OPC. Although the contributions of each agent to oncogenesis in this model are difficult to evaluate, the rapid development of two retrovirus-induced pulmonary diseases in experimentally inoculated lambs suggests an etiologic or pathogenetic synergism between these two members of the family Retroviridae.

Sheep pulmonary adenomatosis: a unique model of retrovirus-associated lung cancer.

Sheep pulmonary adenomatosis (SPA) is a contagious bronchiolo-alveolar carcinoma of sheep associated with an exogenous type D/B retrovirus known as jaagsiekte sheep retrovirus (JSRV). SPA represents a unique model for lung cancer, and studies on its aetiopathogenesis can provide further insight into the mechanisms of epithelial neoplasms.

Natural history of JSRV in sheep.

Ovine pulmonary adenocarcinoma (OPA) is a contagious lung tumour of sheep and, rarely, goats that arises from two types of secretory epithelial cell that retain their luxury function of surfactant synthesis and secretion. It is classified as a low-grade adenocarcinoma and is viewed as a good model for epithelial neoplasia because of its morphological resemblance to the human lung tumour, bronchioloalveolar adenocarcinoma. OPA is present in most of the sheep rearing areas of the globe and, in affected flocks, tumours are present in a high proportion of sheep. OPA is associated with the ovine retrovirus, jaagsiekte sheep retrovirus (JSRV), and is transmissible only with inocula that contain JSRV. All sheep contain JSRV-related endogenous viruses, but JSRV is an exogenous virus that is associated exclusively with OPA. JSRV is detected consistently in the lung fluid, tumour and lymphoid tissues of sheep affected by both natural and experimental OPA or unaffected in-contact flockmates and never in sheep from unaffected flocks with no history of the tumour. JSRV replicates principally in the epithelial tumour cells, but also establishes a disseminated infection of several lymphoid cell types, including peripheral blood leukocytes (PBLs). Longitudinal studies in flocks with endemic OPA have revealed JSRV in PBLs before the onset of clinical OPA and even in the absence of discernible lung tumour. The prevalence of JSRV infection is 40%-80%, although only 30% of sheep appear to develop OPA lesions. A unique feature of OPA is the absence of a specific humoral immune response to JSRV, despite the highly productive infection in the lungs and the disseminated lymphoid infection. This feature is associated with reduced responsiveness to some mitogens, although the phenotypic profile of the peripheral blood remains unaltered. The reduced response is an early and sustained event during infection and may indicate that the failure of infected sheep to produce specific antibodies to JSRV is a direct consequence of infection.

Pathology of ovine pulmonary adenocarcinoma.

Clinical, gross pathology, histopathology and electron microscopy of the ovine pulmonary adenocarcinoma (OPA, jaagsiekte) either natural or experimentally induced in sheep, goat and moufflon are described. OPA is caused by an oncogenic betaretrovirus,jaagsiekte sheep retrovirus (JSRV). Most natural cases of OPA appear in animals 1-4 years old. There is no evidence of sex or breed susceptibility. Sheep affected by OPA show an afebrile respiratory illness associated with loss of weight. A very characteristic clinical sign is moist rales caused by the accumulation of fluid in the respiratory airways which is discharged from the nostrils when the head is lowered. Gross lesions are confined to the lungs but occasionally thoracic or extrathoracic structures are also affected. Two pathologic forms of OPA are currently recognized, classical and atypical. In classical forms the neoplastic lesions occurs particularly in the cranioventral parts of all lung lobes. They are diffuse or nodular, light grey or light purple in colour. On the cut surface the tumour is moist, and frothy fluid may pour from the airways on slight pressure. Atypical forms tend to be more nodular in both early and advanced tumours. They are pearly white in colour, very hard in consistency, very well demarcated from the surrounding parenchyma and their surface is dry. Histology of the lung sections reveals the presence of several foci of epithelial cell neoplastic proliferation in both alveolar or bronchiolar regions. The tumours, derived from type II pneumocytes and Clara cells, proliferate into mostly papillary but also acinar or occasionally solid growths. The tumour generally shows a benign histological pattern but intra- and extrathoracic metastases have been detected in some cases. Several considerations suggest that the tumour should be classified as an adenocarcinoma of the lung. The histology of atypical OPA is similar to that of the classical disease, with an increase in the stromal reaction accompanying the epithelial proliferations. Pathological features of OPA induced experimentally in sheep, or of OPA in goats and moufflon are similar to those described in sheep. Detailed electron microscopy of tumour material confirms that type II pneumocytes and Clara bronchiolar epithelial cells are the origin of the neoplasia. Also included in this chapter is a description of the morphology of the viral particles associated with OPA.

Enzootic nasal adenocarcinoma of sheep and goats.

Enzootic nasal adenocarcinoma is a contagious tumour of the mucosal nasal glands affecting young adult sheep or goats. The disease occurs naturally in all continents except Australia and New Zealand. Clinical signs include continuous nasal discharge, respiratory distress, exophthalmos and skull deformations. The tumour is classified histologically as a low-grade adenocarcinoma. Nasal glands of both respiratory and olfactory muosal glands seem to be the origin of the neoplasia. It has been experimentally transmitted in sheep and goats using either tumour extracts or concentrated nasal fluids. Two distinct retroviruses are implicated in the aetiology of the neoplasia one in sheep (ONAV) and one in goats (CNAV). We suggest that jaagsiekte sheep retrovirus (JSRV), ONAV, CNAV, and their endogenous counterparts represent a unique family of retroviruses. The similarities between these viruses suggests that any control strategies, including vaccination, may be appropriate to both diseases. The differences, however, represent a unique resource for delineating the function of individual regions of the virus. It is intriguing that whilst ONAV and CNAV appear to be as different to each other as they are to JSRV, that they have very similar disease pathologies, distinct from that of OPA. Additionally, all three exogenous viruses manage to avoid instigating any apparent immune response. Whether this is indeed a result of tolerance induced by the endogenous counterparts or whether the viruses themselves have unique immunosuppressive properties will be an important finding.

Genetic diversity of Mycobacterium avium subspecies paratuberculosis isolates from goats detected by pulsed-field gel electrophoresis.

Paratuberculosis in goats occurs worldwide causing considerable economic losses mainly due to reduced milk production. Nowadays, there is still relatively little knowledge about the epidemiology of this disease in goats, and only a few epidemiological studies have been carried out in goats naturally infected with Mycobacterium avium subspecies paratuberculosis (M. a. paratuberculosis). The objective of this study was to characterize forty four clinical caprine isolates of M. a. paratuberculosis by different molecular techniques (pulsed-field gel electrophoresis [PFGE], restriction fragment length polymorphism analysis coupled with hybridization to IS900, and IS1311 polymerase chain reaction-restriction enzyme analysis) to determine the most useful technique for molecular typing of caprine isolates, as well as to disclose the genetic variation amongst caprine isolates and the relationship with strains isolated from other animal species. PFGE was found to be the most discriminative technique identifying a total of 13 'multiplex' PFGE profiles, ten of which were novel profiles found only in caprine isolates to date. All isolates were genotyped as Type II strains, except two isolates that resembled the intermediate group referred as Type III.

An influx of macrophages is the predominant local immune response in ovine pulmonary adenocarcinoma.

Infection with a retrovirus, Jaagsiekte sheep retrovirus (JSRV), causes ovine pulmonary adenocarcinoma (OPA). The excess production of surfactant proteins by alveolar tumour cells results in increased production of pulmonary fluid, which is characteristically expelled through the nostrils of affected sheep. The immune response to JSRV and the tumour is poorly understood: no JSRV-specific circulating antibodies or T cells have been detected to date. The aim of the present study was to obtain phenotypic evidence for a local immune response in OPA lungs. Specific-pathogen free lambs were infected intratracheally with JSRV. When clinical signs of OPA were apparent, the lungs were removed at necropsy and immunohistochemistry (IHC) was performed on lung sections using a panel of mouse anti-sheep mAbs. No influx of dendritic cells, B cells, CD4, CD8 or gammadelta T cells was seen in the neoplastic nodules or in their periphery. MHC Class II-positive cells were found intratumourally, peritumourally and in the surrounding alveolar lumina. In the tumours, many of these cells were shown to be fibroblasts and the remainder were likely to be mature macrophages. In the alveolar lumen, the MHC Class II-positive cells were CD14-positive and expressed high levels of IFN-gamma. They appeared to be immature monocytes or macrophages which then differentiated to become CD14-negative as they reached the periphery of the tumours. A high level of MHC Class I expression was detected on a range of cells in the OPA lungs but the tumour nodules themselves contained no MHC Class I-positive cells. On the basis of these findings, it is proposed that the lack of an effective immune response in OPA could result from a mechanism of peripheral tolerance in which the activity of the invading macrophages is suppressed by the local environment, possibly as a consequence of the inhibitory properties of the surfactant proteins.

Complete sequence of the gene encoding the bacterioferritin subunit of Mycobacterium avium subspecies silvaticum.

A gene encoding the bacterioferritin subunit (Bfr) of Mycobacterium avium (Ma) subspecies silvaticum has been cloned, sequenced and expressed. The 477-bp open reading frame codes for 159 amino acids, which were shown to share up to 92% identity with the Bfr of five bacterial genera. The recombinant Bfr exhibits serological cross-reactivity with Ma paratuberculosis antigen D, a protein of approx. 20 kDa in cell lysates of Ma paratuberculosis and Ma silvaticum and a protein of 20-22 kDa in sonicates of M. leprae.

An accessory open reading frame (orf-x) of jaagsiekte sheep retrovirus is conserved between different virus isolates.

Jaagsiekte sheep retrovirus (JSRV) is the etiological agent of a contagious lung tumour of sheep known as sheep pulmonary adenomatosis (syn: ovine pulmonary carcinoma, jaagsiekte). JSRV exhibits a simple genetic organization, characteristic of the type D and type B retroviruses, with the canonical retroviral sequences gag, pro, pol and env encoding the structural proteins of the virion. An additional open reading frame (orf-x), of approximately 500 bp overlapping pol, is present in the only two complete sequences of JSRV published to date. Since very little information is available on the biology of JSRV it is important to establish if orf-x is conserved between different virus isolates. In this study we analysed the orf-x region of JSRV isolates collected from the United Kingdom, Italy, Spain and South Africa. In addition we also analysed the presence of orf-x in JSRV-related endogenous sequences (enJSRVs) present in the sheep genome. Orf-x was highly conserved in all the exogenous isolates (n=10) and in most of the endogenous sequences (n=8). Thus orf-x may be an accessory gene of JSRV and haves a biological function which might be advantageous to JSRV. Phenetic analysis conducted on the complete orf-x nucleotide sequences seems to highlight the presence of three distinct groups statistically well supported by bootstrapping: i) exogenous JSRV sequence from the UK; ii) exogenous JSRV sequences from Southern Europe and iii) the exogenous South African strain plus all the endogenous sequences analyzed and collected from Australia, Italy, UK and South Africa.

NPY Y1 receptors exert opposite effects on corticotropin releasing factor and noradrenaline overflow from the rat hypothalamus in vitro.

Neuropeptide Y (NPY), corticotropin releasing factor (CRF) and noradrenaline play important roles in the regulation of a number of endocrine and autonomic functions. NPY is co-localised with noradrenaline in the central nervous system and has been observed to modulate noradrenaline release. Recent morphological and physiological studies also support co-modulatory interactions between NPY and CRF. Earlier in vivo studies in our laboratory showed a potentiation of K(+)-stimulated noradrenaline release following NPY administration, possibly due to an NPY Y1 receptor mechanism. In this study, in vitro superfusion techniques were established to simultaneously monitor the release of endogenous noradrenaline and CRF from the hypothalamus of adult rats and to examine the direct neuromodulatory action of NPY on the overflow of CRF and noradrenaline. Administration of 0.10 microM NPY significantly increased CRF overflow to 395% basal levels and reduced hypothalamic noradrenaline overflow to 61% of basal levels. These effects were blocked by prior administration of the NPY Y1 receptor antagonist GR231118. Thus, this study suggests that NPY, working through a Y1 receptor, has dual and opposing effects on CRF and noradrenaline overflow in vitro.

Analysis of multidrug-resistant Mycobacterium bovis from three clinical samples from Scotland.

OBJECTIVE: To determine whether multidrug-resistant (MDR) strains of Mycobacterium bovis isolated from patients in Scotland were genotypically related. DESIGN: Genotypes of MDR strains were determined using three molecular fingerprinting techniques: pulsed-field gel electrophoresis (PFGE), spoligotyping and restriction fragment length polymorphism (RFLP). PFGE profiles were also obtained for all medical and veterinary isolates occurring in Scotland in 1997-1998. RESULTS: MDR strains showed individual Dra I PFGE profiles. Case III/98 had a profile represented in both veterinary and medical populations, Case I/94 had a profile observed in medical but not veterinary isolates, and Case II/98 had a profile unique to this study. Afl II PFGE discriminated the resistant strains. Spoligotyping grouped Cases I/94 and II/98 (ST-134). Case III/98 had a spoligotype ST-140, which is commonly observed in veterinary isolates. Similarly, DRr-RFLP analysis grouped cases I/94 and II/98, whereas Case III/98 had a common veterinary profile. DRX(PGRS) RFLP gave three unique profiles. CONCLUSION: Three resistant strains were discriminated by PFGE and DRX(PGRS) RFLP, indicating that the three strains are not related in an epidemiologically relevant time scale. However, Cases I/94 and II/98 were more closely linked by spoligotyping and DRr-RFLP data. PFGE and DRr-RFLP linked Case III/98 profiles to the most common veterinary isolate.

Tribal differences in perception of tuberculosis: a possible role in tuberculosis control in Arusha, Tanzania.

SETTING: Arusha, Tanzania. OBJECTIVE: To determine tribal differences in knowledge and practices that might influence tuberculosis control. METHOD: Twenty-seven villages were selected randomly out of 242 villages in four districts. In each village, a general and a livestock keeping group were selected at random. The households were home-visited and 426 family members were interviewed. RESULTS: On average, 40% of respondents practised habits that might expose them to both bovine and human tuberculosis. The Barabaig tribe had a significantly higher number of respondents (50%, chi2(2) = 5.1, P = 0.024) who did not boil milk. Eating uncooked meat or meat products was practised by 17.9% of all respondents. The habit was practised more by Iraqw (21.1%, chi2(2) = 6.9, P = 0.008) and Barabaig (31.6%, chi2(2) = 5.6, P = 0.016) than other tribes. About 75% of the respondents had a poor knowledge of tuberculosis. CONCLUSION: All tribes had habits and beliefs that might expose them to both bovine and human tuberculosis. The Iraqw and Barabaig tribes practised such habits more than other tribes. Knowledge of tuberculosis was limited in all tribes.

Isolation of Mycobacterium bovis from human cases of cervical adenitis in Tanzania: a cause for concern?

SETTING: Pastoralist communities in the Northern and Southern zones of Tanzania. DESIGN: Observational study. OBJECTIVES: To determine the involvement of Mycobacterium bovis in tuberculosis cases presenting at tuberculosis (TB) clinics in rural areas in these zones. METHODS: A total of 149 tuberculosis cases identified on the bases of clinical manifestation were sampled. Appropriate specimens were cultured on two Löweinstein Jensen slants with respectively glycerol and pyruvate added. Forty-one isolates were cultured and subjected to biochemical typing. RESULTS: Overall, 31 (70.5%) of the mycobacterial isolates recovered from all forms of tuberculosis were identified as M. tuberculosis, seven (16.0%) were identified as M. bovis, and six (13.6%) were other mycobacterial species. There was a significantly higher isolation rate (P < 0.05) of M. bovis among strains recovered from extra-pulmonary (26.8%) than pulmonary tuberculosis samples (4.3%). CONCLUSION: Based on these findings, it is imperative that M. bovis be considered as a pathogen of concern to people living in rural areas of Tanzania. Further work is required to establish a zoonotic link between cattle and the people in these communities who rear them.

The role of livestock keeping in tuberculosis trends in Arusha, Tanzania.

SETTING: Arusha, Tanzania. OBJECTIVE: To assess risk factors that might influence TB control in the general population and in livestock-keepers. METHODS: Of 242 villages in four districts, 27 were selected randomly. In each village, a general and a livestock-keeping group were selected at random. The households were home-visited and 426 family members were interviewed. RESULTS: On average, three-quarters of households practised at least one risk activity for transmission of zoonotic tuberculosis, and respondents had poor knowledge about tuberculosis. In the livestock-keeping group, the risks of having a tuberculosis patient in the family were determined by poor ventilation (OR 2.6, 95% CI 1.1-6.5), confining livestock indoors with people (OR 2.3, 95%CI 1.1-5.0) and multiple determinants including poor ventilation (OR 13.5, 95% CI 2.5-71.7). Risk activities and the risks of having a tuberculosis patient in a family were significantly higher in the livestock-keeping group. CONCLUSIONS: The respondents had limited knowledge about tuberculosis, and the households had practices that posed potential risks for both human and bovine tuberculosis infection. Poor ventilation and confining livestock indoors were associated with tuberculosis spread in the households. These risks were observed more in the livestock-keeping group than in the general population group.

In vitro studies of endogenous noradrenaline and NPY overflow from the rat hypothalamus during maturation and ageing.

Marked changes in brain monoamine content and NPY content occur during maturation and ageing. Earlier in vivo studies in our laboratory have reported blunted K+ stimulated noradrenaline release and reduced NPY overflow in aged animals using microdialysis and push pull techniques. In this study, in vitro superfusion techniques were established to measure endogenous noradrenaline, NPY, DOPAC and 5-HIAA overflow from the hypothalamus of 1, 5 and 16 month old Sprague-Dawley rats. A period of high K+ (56 mM) stimulation was carried out to elicit maximal release. Basal noradrenaline overflow was similar in all age groups of rats and during K+-induced depolarisation similar 3-4 fold increases were observed. On the other hand, basal and K+ stimulated NPY overflow were significantly greater in the adult rats compared to 1 month and 16 month old rats. Despite differences in absolute NPY overflow, the relative increase over resting was not significantly different across age groups. The molar quantities of hypothalamic NPY overflow at rest and under K+ stimulated conditions were three orders of magnitude lower than noradrenaline. Results of these studies suggest that both NPY and noradrenaline can be released from a similar hypothalamic pool. Basal and K+-evoked DOPAC and 5-HIAA overflow were similar between the 3 age groups. Thus the overflow of hypothalamic noradrenaline, DOPAC and 5-HIAA under in vitro conditions was not altered from 1 to 16 months. In contrast, 5 month old rats had significantly higher NPY overflow than the other age groups (P < 0.05), consistent with a reported decline in NPY content with advanced age. Hypothalamic noradrenaline overflow was not affected by ageing, suggesting that a selective loss of NPY in the arcuo-PVN projection, or other projections to the hypothalamus with ageing may contribute to the reduction in NPY overflow in aged rats.