Somatic Volume Research Articles

Plastic rearrangement of basal forebrain parvalbumin-immunoreactive neurons in the kainite model of epilepsy.

Temporal lobe epilepsy (TLE) is the most prevalent form of epilepsy, through the neuronal mechanisms of this syndrome remain elusive. In addition to the temporal lobe structures, it was found that the basal forebrain cholinergic cells are also involved in epileptogenesis. However, little is known about the involvement of the basal forebrain GABAergic neurons in epilepsy; despite this, they largely project to the temporal lobe and are crucial for the regulation of the hippocampal circuitry. In this study, we assessed epilepsy-induced changes in parvalbumin (PARV) immunoreactive neurons of the medial septum (MS) and of the magnocellular preoptic nucleus (MCPO) using the kainic acid (KA) model in rats. In addition, we estimated the respective changes in the cholinergic varicosities in the MS, where we observed a significant reduction in the PARV cell number (12849 ± 2715 vs. 9372 ± 1336, p = .029) and density (16.2 ± 2.62 vs. 10.5 ± 1.00 per .001 mm3, p =.001), and an increase in the density of cholinergic varicosities (47.9 ± 11.1 vs. 69.4 ± 17.8 per 30,000 µm2, p =.036) in KA-treated animals. In the MCPO, these animals showed a significant increase in somatic volume (827.9 ± 235.2 µm3 vs. 469.9 ± 79.6 µm3, p = .012) and total cell number (2268.6 ± 707.1 vs. 1362.4 ± 262.0, p =.028). These results show that the basal forebrain GABAergic cell populations undergo numerical and morphological changes in epileptic animals, which may contribute to an increased vulnerability of brain circuits to epilepsy and epilepsy-related functional impairments.

Effects of aging on the cholinergic innervation of the rat ventral tegmental area: A stereological study

Dopamine neurons in the ventral tegmental area (VTA) play a main role in processing both rewarding and aversive stimuli, and their response to salient stimuli is significantly shaped by afferents originating in the brainstem cholinergic nuclei. Aging is associated with a decline in dopaminergic activity and reduced response to positive reinforcement. We have used stereological techniques to examine, in adult and aged rats, the dopaminergic neurons and the cholinergic innervation of the VTA, and the cholinergic populations of the pedunculopontine tegmental (PPT) and laterodorsal tegmental (LDT) nuclei, which are the only source of cholinergic inputs to the VTA. In the VTA, there were no age-related variations in the number and size of tyrosine hydroxylase (TH)-immunoreactive neurons, but the density of cholinergic varicosities was reduced in aged rats. The total number of choline acetyltransferase (ChAT)-immunoreactive neurons in the PPT and LDT was unchanged, but their somas were hypertrophied in aged rats. Our results suggest that dysfunction of the cholinergic system might contribute for the age-associated deterioration of the brain reward system.

Chronic intermittent ethanol and lipopolysaccharide exposure differentially alter Iba1-derived microglia morphology in the prelimbic cortex and nucleus accumbens core of male Long-Evans rats.

Accumulating evidence has linked pathological changes associated with chronic alcohol exposure to neuroimmune signaling mediated by microglia. Prior characterization of the microglial structure-function relationship demonstrates that alterations in activity states occur concomitantly with reorganization of cellular architecture. Accordingly, gaining a better understanding of microglial morphological changes associated with ethanol exposure will provide valuable insight into how neuroimmune signaling may contribute to ethanol-induced reshaping of neuronal function. Here we have used Iba1-staining combined with high-resolution confocal imaging and 3D reconstruction to examine microglial structure in the prelimbic (PL) cortex and nucleus accumbens (NAc) in male Long-Evans rats. Rats were either sacrificed at peak withdrawal following 15days of exposure to chronic intermittent ethanol (CIE) or 24hr after two consecutive injections of the immune activator lipopolysaccharide (LPS), each separated by 24hr. LPS exposure resulted in dramatic structural reorganization of microglia in the PL cortex, including increased soma volume, overall cellular volume, and branching complexity. In comparison, CIE exposure was associated with a subtle increase in somatic volume and differential effects on microglia processes, which were largely absent in the NAc. These data reveal that microglial activation following a neuroimmune challenge with LPS or exposure to chronic alcohol exhibits distinct morphometric profiles and brain region-dependent specificity.

Effects of chronic alcohol consumption, withdrawal and nerve growth factor on neuropeptide Y expression and cholinergic innervation of the rat dentate hilus

Several studies have demonstrated the vulnerability of the hippocampal formation (HF) to chronic alcohol consumption and withdrawal. Among the brain systems that appear to be particularly vulnerable to the effects of these conditions are the neuropeptide Y (NPY)-ergic and the cholinergic systems. Because these two systems seem to closely interact in the HF, we sought to study the effects of chronic alcohol consumption (6months) and subsequent withdrawal (2months) on the expression of NPY and on the cholinergic innervation of the rat dentate hilus. As such, we have estimated the areal density and the somatic volume of NPY-immunoreactive neurons, and the density of the cholinergic varicosities. In addition, because alcohol consumption and withdrawal are associated with impaired nerve growth factor (NGF) trophic support and the administration of exogenous NGF alters the effects of those conditions on various cholinergic markers, we have also estimated the same morphological parameters in withdrawn rats infused intracerebroventricularly with NGF. NPY expression increased after withdrawal and returned to control values after NGF treatment. Conversely, the somatic volume of these neurons did not differ among all groups. On other hand, the expression of vesicular acetylcholine transporter (VAChT) was reduced by 24% in ethanol-treated rats and by 46% in withdrawn rats. The administration of NGF to withdrawn rats increased the VAChT expression to values above control levels. These results show that the effects of prolonged alcohol intake and protracted withdrawal on the hilar NPY expression differ from those induced by shorter exposures to ethanol and by abrupt withdrawal. They also suggest that the normalizing effect of NGF on NPY expression might rely on the NGF-induced improvement of cholinergic neurotransmission in the dentate hilus.

Developmental changes in the morphology of mouse hypoglossal motor neurons.

Hypoglossal motor neurons (XII MNs) innervate tongue muscles important in breathing, suckling and vocalization. Morphological properties of 103 XII MNs were studied using Neurobiotin™ filling in transverse brainstem slices from C57/Bl6 mice (n = 34) from embryonic day (E) 17 to postnatal day (P) 28. XII MNs from areas thought to innervate different tongue muscles showed similar morphology in most, but not all, features. Morphological properties of XII MNs were established prior to birth, not differing between E17–18 and P0. MN somatic volume gradually increased for the first 2 weeks post-birth. The complexity of dendritic branching and dendrite length of XII MNs increased throughout development (E17–P28). MNs in the ventromedial XII motor nucleus, likely to innervate the genioglossus, frequently (42 %) had dendrites crossing to the contralateral side at all ages, but their number declined with postnatal development. Unexpectedly, putative dendritic spines were found in all XII MNs at all ages, and were primarily localized to XII MN somata and primary dendrites at E18–P4, increased in distal dendrites by P5–P8, and were later predominantly found in distal dendrites. Dye-coupling between XII MNs was common from E18 to P7, but declined strongly with maturation after P7. Axon collaterals were found in 20 % (6 of 28) of XII MNs with filled axons; collaterals terminated widely outside and, in one case, within the XII motor nucleus. These results reveal new morphological features of mouse XII MNs, and suggest that dendritic projection patterns, spine density and distribution, and dye-coupling patterns show specific developmental changes in mice.

Chronic alcohol consumption leads to neurochemical changes in the nucleus accumbens that are not fully reversed by withdrawal

Neuropeptide Y (NPY)- and acetylcholine-containing interneurons of the nucleus accumbens (NAc) seem to play a major role in the rewarding effects of alcohol. This study investigated the relationship between chronic alcohol consumption and subsequent withdrawal and the expression of NPY and acetylcholine in the NAc, and the possible involvement of nerve growth factor (NGF) in mediating the effects of ethanol. Rats ingesting an aqueous ethanol solution over 6months and rats subsequently deprived from ethanol during 2months were used to estimate the total number and the somatic volume of NPY and cholinergic interneurons, and the numerical density of cholinergic varicosities in the NAc. The tissue content of choline acetyltransferase (ChAT) and catecholamines were also determined. The number of NPY interneurons increased during alcohol ingestion and returned to control values after withdrawal. Conversely, the number and the size of cholinergic interneurons, and the amount of ChAT were unchanged in ethanol-treated and withdrawn rats, but the density of cholinergic varicosities was reduced by 50% during alcohol consumption and by 64% after withdrawal. The concentrations of dopamine and norepinephrine were unchanged both during alcohol consumption and after withdrawal. The administration of NGF to withdrawn rats significantly increased the number of NPY-immunoreactive neurons, the size of cholinergic neurons and the density of cholinergic varicosities. Present data show that chronic alcohol consumption leads to long-lasting neuroadaptive changes of the cholinergic innervation of the NAc and suggest that the cholinergic system is a potential target for the development of therapeutic strategies in alcoholism and abstinence.

Nerve growth factor retrieves neuropeptide Y and cholinergic immunoreactivity in the nucleus accumbens of old rats

The nucleus accumbens (NAc) contains high levels of neuropeptide Y (NPY), which is involved in the regulation of functions and behaviors that deteriorate with aging. We sought to determine if aging alters NPY expression in this nucleus and, in the affirmative, if those changes are attributable to the cholinergic innervation of the NAc. The total number and the somatic volume of NPY- and choline acetyltransferase-immunoreactive neurons, and the density of cholinergic varicosities were estimated in the NAc of adult (6 months old) and aged (24 months old) rats. In aged rats, the number of NPY neurons was reduced by 20% and their size was unaltered. The number of cholinergic neurons and the density of the cholinergic varicosities were unchanged, but their somas were hypertrophied. Nerve growth factor administration to aged rats further increased the volume of cholinergic neurons, augmented the density of the cholinergic varicosities, and reversed the age-related decrease in the number of NPY neurons. Our data show that the age-related changes in NPY levels in the NAc cannot be solely ascribed to the cholinergic innervation of the nucleus.

Neuronal somatic volume of posteroventral medial amygdala cells from males and across the estrous cycle of female rats

The posteroventral medial amygdala (MePV) is a brain area where gonadal hormones have neurotrophic effects in rats. The aim of the present study was to estimate the MePV neuronal somatic volume from males and diestrus, proestrus and estrus female Wistar rats (n=5 in each group) in an attempt to identify a possible sexual dimorphism in this parameter. The effect of laterality was also evaluated. The brains of adult animals were sectioned (1 microm), stained with 1% toluidine blue and serial-section reconstructions of each neuronal cell body were obtained. Images from both left and right MePV were studied and the somatic volume was estimated using the Cavalieri method in combination with the point counting technique. Results were compared according to sex and phase of the estrous cycle using a two-way ANOVA for repeated measures followed by the least significance difference test. Mean neuronal somatic volume showed a statistical difference among groups and the post hoc comparisons revealed that males present higher values than females in proestrus and estrus (p<0.05). On the other hand, neither a laterality effect (p=0.6) nor an interaction between groups and laterality (p=0.4) were found. Our results indicate that cell body volume in the MePV is distinct when comparing males to females in the different phases of the estrous cycle. Through dynamic changes modulated by sex steroids, it is likely that this morphological plasticity within the MePV may be affecting the functioning of local neurons and their integrated roles in neural circuits relevant for neuroendocrine control and reproductive behaviors.

Influence of sex and estrous cycle, but not laterality, on the neuronal somatic volume of the posterodorsal medial amygdala of rats

The aim of the present study was to measure the cell body volume of neurons from the posterodorsal subnucleus of the medial amygdala (MePD) of adult male ( n = 5) and diestrus, proestrus and estrus female ( n = 4–5 in each group) rats to reveal a possible sexual dimorphism, estrous cycle variations and laterality in this morphological parameter. The brains of adult Wistar rats were sectioned (1 μm), stained with 1% toluidine blue and the stereological estimation of neuronal soma volume of both sides of MePD was realized using the Cavalieri method and the technique of point counting. Data were compared by a two-way ANOVA for repeated measures and the least significance difference post hoc test. In the MePD, mean neuronal somatic volume showed a statistical difference among groups ( p = 0.005), but neither an effect of laterality ( p = 0.33) nor interactions between groups and laterality ( p = 0.78) were found. Post hoc test showed that males (mean ± S.E.M., 2075.67 ± 135.79 μm 3) have larger mean neuronal somatic volume compared to females in proestrus (1503.30 ± 44.46 μm 3) and in estrus (1616.69 ± 71.49 μm 3, p < 0.05 in both cases), but not in diestrus (1940.78 ± 129.68 μm 3, p > 0.05). Moreover, diestrus females displayed larger mean neuronal somatic volume than proestrus female rats ( p < 0.05). It is suggested that neuronal somatic volume is another sexually dimorphic finding in the MePD, for which it is relevant to set apart the different phases of the estrous cycle to reveal the presence of gonadal hormones effects in the rat MePD neurons.

Nerve growth factor restores the expression of vasopressin and vasoactive intestinal polypeptide in the suprachiasmatic nucleus of aged rats

Aging leads to a decrease in the number of neurons expressing vasopressin (VP) and vasoactive intestinal polypeptide (VIP) in the suprachiasmatic nucleus (SCN) of the rat. Similar results were observed following prolonged alcohol consumption and withdrawal. In the latter circumstances, the administration of nerve growth factor (NGF) restored the synthesis and expression of those neuropeptides despite the absence of TrkA receptors in SCN neurons. Thus, we decided to test whether the administration of NGF would improve the expression of neuropeptides in the SCN of aged rats. For this purpose, NGF was delivered intraventricularly to aged rats over a period of 14 days. The somatic volume and the total number of VP- and VIP-immunostained SCN neurons were estimated by applying stereological methods. No age-related variations were found regarding the volume of the neuronal cell bodies. Yet, a striking reduction in the number of VP- and VIP-immunoreactive neurons was detected in aged animals and found to be completely retrieved by NGF. This finding shows that exogenous NGF administered to aged rats restores the neurochemical phenotype of the SCN. This might occur either through direct signaling of SCN neurons via p75 NTR or through enhancement of the cholinergic input to the SCN.

The effects of nerve growth factor upon the neuropeptide content of the suprachiasmatic nucleus of rats withdrawn from ethanol are mediated by the nucleus basalis magnocellularis

It has been previously shown that withdrawal from alcohol decreases the synthesis and expression of vasopressin (VP) and vasoactive intestinal polypeptide (VIP) in the suprachiasmatic nucleus (SCN), and that the infusion of NGF over 1 month completely restores these changes. Because SCN neurons do not express TrkA, NGF might have exerted its effects either through direct signalling of the neurons via p75NTR or by enhancing the activity of the cholinergic afferents to the SCN, which arise from the nucleus basalis magnocellularis (NBM). The observation that the infusion of NT-3 to withdrawn rats does not elicit any change in neuropeptide expression in the SCN suggests that ACh might be implicated in this process, a hypothesis that we have attempted to clarify in this study. For this purpose we destroyed, with quinolinic acid, the NBM of rats withdrawn from ethanol and later infused them with NGF over a period of 13 days. The total number and the somatic volume of SCN neurons immunoreactive for VP and VIP were stereologically estimated. No differences were found in the total number of neurons between quinolinic-injected NGF-treated withdrawn animals and intact withdrawn rats. However, the somatic volume of SCN neurons from quinolinic-injected animals was significantly reduced relative to control and withdrawn rats. The present results unequivocally demonstrate that the trophic effects exerted by NGF upon SCN neurons do not depend on direct neuronal signalling. Instead, they are indirect and, according to our results, NBM neurons, whose axons give rise to a cholinergic projection to the SCN, seem to be essential for eliciting those effects.

NGF and NT-3 exert differential effects on the expression of neuropeptides in the suprachiasmatic nucleus of rats withdrawn from ethanol treatment

Some neurotrophins have the capability of enhancing neuropeptide expression in several regions of the brain. It was also recently shown that NGF, infused over 1 month, offsets the decreased synthesis and expression of vasopressin (VP) and vasoactive intestinal polypeptide (VIP) in the suprachiasmatic nucleus (SCN) of rats submitted to chronic ethanol treatment and withdrawal. In the present study we examined the effectiveness of neutrotrophin-3 (NT-3) in promoting such effects, given that SCN neurons express both the high and the low affinity receptors for this neurotrophin. NT-3 was intraventricularly infused during 10 days to rats withdrawn from prolonged ethanol treatment. The total number, and the mean somatic volume, of VP- and VIP-immunoreactive neurons was compared with the estimates obtained from control rats and withdrawn rats treated with either NGF or cerebrospinal fluid during the same period. The infusion of cerebrospinal fluid and of NT-3 did not prevent the reduction in the number of peptide-producing neurons induced by withdrawal from ethanol treatment. Conversely, NGF infusion increased their number to control levels and led to neuronal hypertrophy. Our results show that, unlike NGF, NT-3 does not display the capacity of enhancing neuropeptide expression in the SCN. Because SCN neurons express the low affinity p75 NTR, which is equally activated by both neurotrophins, our results additionally indicate that the effects of NGF upon SCN neurons are not receptor-mediated. Taken together, our data suggest that indirect mechanisms, rather than direct neutrophin signaling, are likely to mediate the trophic effects exerted by NGF upon SCN neurons.

Stereological evaluation and Golgi study of the sexual dimorphisms in the volume, cell numbers, and cell size in the medial preoptic nucleus of the rat.

The medial preoptic nucleus (MPN) and the sexually dimorphic nucleus of the preoptic area (SDN-POA) stand out as prominent sexually dimorphic cell groups of the rat brain. However, quantitative data on sex-related differences in these nuclei in the adult rat are confined to their volume. We have used stereological methods and Golgi-impregnated material to examine whether, in young adult rats, the sexual dimorphism in the volume of the MPN, including its divisions, and of the SDN-POA, reflect similar differences in the number and size of their neurons. We found that the total number of neurons in all MPN divisions is higher and the mean somatic volume larger in males than in females. In addition, the total dendritic length of MPN neurons is greater, but the dendritic spine density is smaller, in males than in females. Likewise, in the SDN-POA the total number and size of its neurons is greater in males than in females. The sex differences in all quantitative parameters evaluated accounted for the larger volume of the MPN and SDN-POA in males relative to females. In addition, the MPN neuropil also displays sex-related differences in its volume, and these differences closely match those detected for the volume of each MPN division. It deserves to be emphasised that the numerical density of neurons was the only parameter found to be significantly higher in females than in males in all MPN divisions and in the SDN-POA. Our results show that the MPN and the SDN-POA display sex differences in the volume, total number of neurons, and size of neuronal cell bodies and dendritic trees. Furthermore, they also indicate that the neuropil is critical for the establishment of sexual dimorphism in the size of the MPN.

Age-related changes in the volume of somata and organelles of cerebellar granule cells

Because cerebellar granule cells are fixed post-mitotic cells, it is expected that they undergo age-related changes like other neurons. To examine this possibility, a stereological study on granule cells of rat neocerebellar cortex was performed for an age spectrum of 2 to 24 months using eight different age groups. The nucleator method, together with point and intersection counting, was used to obtain primary data; arithmetical calculations determined the secondary data. In the soma, the absolute surface area did not change significantly; the volume did, however, exhibit a significant negative linear trend with age. Excluding dense bodies, the absolute volumes of the cytoplasmic components did not vary significantly. The absolute volume of dense bodies displayed a significant positive linear trend with age. Significant positive correlations were detected between the somatic volume and the absolute volume of either mitochondria or ground substance. It was concluded that granule cells showed a fair degree of morphological stability through 18 months. However, the observed changes warn that accompanying physiological alterations may occur, with putative effects on motor coordination.

Allocation of resources in an European cyprinid during maturation period as measured by nucleic acids

Changes in the nucleic acid levels of different tissues during a regular gonadal maturation were used to investigate the sex-related size differences in Iberian barbel, Barbus sclateri. Gonadal DNA concentration was associated with gonadal development in both sexes. There was a decline in DNA when ovaries were ripe. In contrast, the maximum size of the testes was related with highest gonadal DNA concentrations. Gonadal growth was not related with liver RNA : DNA ratios of male, while it was with female ratios. White muscle DNA concentrations indicate that somatic cell volume decreased during gonadal growth in both sexes. This suggests the translocation of several elements toward the gonad. Moreover, during gonadal maturation period adult barbel showed no change in their somatic growth. However, males apparently were in poorer metabolic condition (lower muscle RNA : DNA ratio) than females. Females may have allocated additional energy to self-maintenance rather than to breeding effort and it might influenced the cumulative annual growth. One might reasonably assume that there is a trade-off between investment in current versus future reproductive success in female. This result was not seen in males.

QUANTITATIVE ALTERATIONS AFTER LONG-TERM ALCOHOL ADMINISTRATION IN THE DORSAL LATERAL GENICULATE NUCLEUS (dLGN IN THE RABBIT (&lt;italic&gt;ORYCTOLAGUS CUNICULUS&lt;/italic&gt;)

The effects of chronic excessive alcohol ingestion on the central nervous system were studied in neurons of the dorsal lateral geniculate nucleus (dLGN) in the rabbit (Oryctolagus cuniculus). In this nucleus, neuron density and other morphometric parameters such as the somatic volume of the cells have been investigated both under normal conditions and under alcohol intoxication. Special attention was given to the possibility that positive somatic heteropycnosis may be a sign of imminent cell death. In addition, the percentage volumes occupied by normal neuronal soma and by affected cells were determined. Continuous alcohol treatment for 6 months reduced the size of certain types of neurons, the alteration being specially intense in areas with an abundance of larger neurons. In these areas more intense signs of somatic heteropycnosis also appeared. We discuss the relationship between the condition of the inhibitory neurons (GABAergic interneurons), the distribution of these in the lower zone of the nucleus, and their greater resistance to the influence of alcohol. This type of stereological analysis is intended to provide a better interpretation of the different degrees of the effects of alcohol, and to give more detailed information about changes at cellular level, both for comparative purposes with other situations and to shed light on the alterations caused by alcoholism.

Distribution of neurons and glia in the visual cortex (area 17) of the adult albino rat: A quantitative description

The neuronal and glial cell composition of the rat visual cortex (area 17) has been determined quantitatively using stereological techniques. The volume numerical densities (number of cells per mm 3 of cortex) of neurons and of the principal glial cell types (astroglia, oligodendroglia, and microglia) were calculated from tangential semithin resin sections spaced at regular intervals 50 μm apart throughout the entire depth of the visual cortex. From measurements of cortical and laminar thickness the separate volume numerical densities of neurons and glial cells were derived for each lamina in the cortex. In addition, the absolute numbers of cells in each lamina under 1 mm 2 of cortical surface were calculated. The mean cortical volume numerical density of neurons was60,020 ± 3840/mm 3 (X ¯± SEM; n = 8), and49,040 ± 2610/mm 3 for the combined glial cell types. Astroglia, oligodendroglia, and microglia were present in a ratio of 6:3:1 respectively. It was determined from neuronal and glial somatic volume estimates that the somata of these cells occupied approximately 13.5% of unit cortical volume, with 81.3% of the unit volume being occupied by cortical neuropil. Using previously published reports that described the laminar composition of neurons in terms of the relative proportions of pyramidal and non-pyramidal cells, the laminar volume numerical densities for these neuronal categories have been derived. In addition, it has been estimated that under 1 mm 2 of cortical surface there are 79,500 pyramidal and 7790 non-pyramidal neurons distributed throughout layers 1–6 of the rat visual cortex.

An HRP study of the cat's spinal respiratory motoneurones during postnatal development

The localization and morphology of spinal respiratory motoneurones (phrenic and intercostal) were studied in the cat by retrograde labelling using Horseradish Peroxidase (HRP), at different stages of postnatal development. At birth, the distribution of the phrenic and intercostal motoneurones in the cervical and thoracic ventral horn, respectively, is similar to that observed in adult animals. At birth, the phrenic and intercartilaginous motoneurone somata have respectively 60% and 40% of their adult volume, appearing much more developed than the motoneurones involved in the motor control of limbs. During postnatal development, the phrenic and intercartilaginous motoneurones undergo a characteristic sagittal elongation without evident modification along their transverse axes. From birth, the ratio of the somatic volume to that of its corresponding motor column markedly decreases inside of the phrenic column compared to the data obtained in the limb's muscle motor columns by other authors. Similar determinations in intercostal motor columns give intermediate values between those obtained from the phrenic column and from the motor system. These results indicate that the motoneurones innervating the respiratory muscles have some specific features of development.

Relationship of somatic cell count and cell volume analysis of goat's milk to intramammary infection with coagulase-negative staphylococci.

The prevalence of intramammary infection in 4 commercial goat herds was studied in conjunction with electronic somatic cell count and volume analysis determined using a Coutler Counter and volume analyser. Neither streptococci nor mycoplasma were isolated from any half and the prevalence of intramammary infection with Staphylococcus aureus ranged from 0 to 3% between herds. For coagulase-negative staphylococci the range from infected halves was 36-71%. There was no significant difference between the mean total microscopic somatic cell count for halves infected with coagulase-negative staphylococci and those free from infection. A similar trend was observed for electronic somatic cell counts although the mean electronic cell count was greater than the mean total microscopic count on the 2 occasions that they were compared. The correlation coefficients between the 2 cell counting methods were 0.86 and 0.94. Between herds there were significant differences in mean electronic somatic cell count, with herd means ranging from 438 x 103 to 1684 x 103 cells/ml. In 2 of the 4 herds studied, milk samples from halves infected with coagulase-negative staphylococci had a significantly higher prevalence of cell volume distributions with a modal cell volume between 65 mu 3 and 100 mu 3. This was attributed to a higher proportion of polymorphonuclear neutrophils. Use of electronic somatic cell count and cell volume analysis were considered of little value in predicting infection caused by coagulase-negative staphylococci as there was a high proportion of false negative and false positive predictions.

Analysis of somatic cell volume distribution as an aid to the diagnosis mastitis.

Electronic somatic cell counts (ESCC) and relative cell volume distributions (RCVD) in milk samples were obtained simultaneously on a Coulter Counter Model TA and their potential assessed, either singly or in combination, for identifying mastitic quarters and cows in 2 herds. Using log (ESCC) alone the probability P m of misclassifying quarters infected with major pathogens was 6.7% for herd 1 and 18.1% for herd 2. The inclusion of RCVD variables with log (ESCC) did not substantially improve the chance of detecting quarters infected with major pathogens. However, the RCVD variables did improve the chance of identifying quarters infected by major and minor pathogens compared to the use of log (ESCC) alone, the P m figures being reduced by about one third. When testing composite quarter samples, where milk from infected quarters was diluted with that from uninfected quarters, the inclusion of RCVD variables decreased the P m compared to the use of log (ESCC) alone. In hered 1 the improvement was from 26.1 to 3.0%, while in herd 2 the effect was less marked, the P m being reduced from 28.6 to 21.6%. However, as the RCVD assessment is provided simultaneously with ESCC, the additional information is obtained at minimal cost.