Age-related increase in the expression of 11ß-hydroxysteroid dehydrogenase type 1 in the hippocampus of male rhesus macaques.

Introduction: The hippocampus is especially susceptible to age-associated neuronal pathologies, and there is concern that the age-associated rise in cortisol secretion from the adrenal gland may contribute to their etiology. Furthermore, because 11ß-hydroxysteroid dehydrogenase type 1 (HSD11B1) catalyzes the reduction of cortisone to the active hormone cortisol, it is plausible that an increase in the expression of this enzyme enhances the deleterious impact of cortisol in the hippocampus and contributes to the neuronal pathologies that underlie cognitive decline in the elderly. Methods: Rhesus macaques were used as a translational animal model of human aging, to examine age-related changes in gene and protein expressions of (HSD11B1/HSD11B1) in the hippocampus, a region of the brain that plays a crucial role in learning and memory. Results: Older animals showed significantly (p < 0.01) higher base-line cortisol levels in the circulation. In addition, they showed significantly (p < 0.05) higher hippocampal expression of HSD11B1 but not NR3C1 and NR3C2 (i.e., two receptor-encoding genes through which cortisol exerts its physiological actions). A similar age-related significant (p < 0.05) increase in the expression of the HSD11B1 was revealed at the protein level by western blot analysis. Discussion: The data suggest that an age-related increase in the expression of hippocampal HSD11B1 is likely to raise cortisol concentrations in this cognitive brain area, and thereby contribute to the etiology of neuropathologies that ultimately lead to neuronal loss and dementia. Targeting this enzyme pharmacologically may help to reduce the negative impact of elevated cortisol concentrations within glucocorticoid-sensitive brain areas and thereby afford neuronal protection.

Acute responses of stevia and d-tagatose intake on metabolic parameters and appetite/satiety in insulin resistance.

OBJECTIVE: To examine the effects of d-tagatose or stevia preloads on carbohydrate metabolism markers after an oral glucose load, as well as subjective and objective appetite in women with insulin resistance (IR). RESEARCH DESIGN AND METHODS: Randomized controlled crossover study. Women with IR without T2DM (n = 33; aged 23.4 ± 3.8; BMI 28.1 ± 3.4 kg × m-2) underwent three oral glucose loads (3 h each) on three different days. Ten min before oral glucose load, volunteers consumed a preload of 60 mL water (control), 60 mL water with stevia (15.3 mg), or d-tagatose (5000 mg). Serum glucose and C-peptide were evaluated at -10, 30-, 60-, 90-, 120-, and 180-min. Subjective appetite was determined with a visual analog scale. Food intake was measured at ad libitum buffet after 180 min. RESULTS: C-peptide iAUC was significantly higher for stevia (median (IQR): 1033 (711-1293) ng × min × L-1) vs. d-tagatose (794 (366-1134) ng × min × L-1; P = 0.001) or control (730 (516-1078) ng × min × L-1; P = 0.012). At 30- and 60-min serum glucose was higher for stevia vs other conditions (P < 0.01). Volunteers reported greater satiety for stevia and d-tagatose vs. control at 60 min and greater desire to eat for stevia vs. control at 120- min (all P < 0.05). Objective appetite did not vary by condition (P = 0.06). CONCLUSIONS: Our findings suggest that these NNS are not inert. Stevia intake produced an acute response on C-peptide release while increased serum glucose at earlier times. It is possible that NNS affects subjective but not objective appetite. This trial is registered at clinicaltrials.gov as NCT04327245. CLINICAL TRIAL REGISTRY: NCT04327245.

Lack of effect of short-term DHEA supplementation on the perimenopausal ovary†.

Dehydroepiandrosterone (DHEA) hormonal supplementation can improve oocyte quality in women with diminished ovarian function. However, it is unclear whether DHEA supplementation can also enhance ovarian function during the perimenopause (i.e., when the number of follicles in the ovary has undergone a marked reduction). To address this question, we examined the impact of 2.5-months of daily 5-mg oral DHEA supplementation on the number of ovarian follicles and the concentration of anti-Müllerian hormone (AMH) in perimenopausal rhesus macaques. Like women, these long-lived nonhuman primates have ~ 28-day menstrual cycles and eventually undergo menopause. They also show similar age-related neuroendocrine changes, including a marked decrease in circulating concentrations of DHEA and DHEA sulfate (DHEAS). Our experimental design involved the following three groups of animals (N = 6 per group): Young adult (mean age = 11.6 years), Old control (mean age = 23.1 years), and Old DHEA-treated (mean age = 23.5 years). Histological examination of the ovaries revealed a significant age-related decrease in the mean number of primordial follicles despite DHEA supplementation. Moreover, AMH concentrations within the ovaries and circulation, assessed by Western analysis and ELISA, respectively, showed significant age-related decreases that were not attenuated by DHEA supplementation. Taken together, these results fail to show a clear effect of short-term physiological DHEA supplementation on the perimenopausal ovary. However, they do not exclude the possibility that alternative DHEA supplementation paradigms (e.g., involving an earlier start date, longer duration and using pharmacological doses) may extend reproductive potential during aging.

Effect of different housing systems (single and group penning) on the health and welfare of commercial female rabbits.

In recent decades, concern about rabbit welfare and sustainability has increased. The housing system is a very important factor for animal welfare. However, information about how different available housing types for female rabbits affect their health status is scarce, but this is an important factor for their welfare. Hence, the objective of this study was to evaluate the health status of female rabbits in five common housing systems: three different single-housing systems with distinct available surfaces and heights; a single-housing system with a platform; a collective system. Female rabbits in the collective and platform cages had greater cortisol concentrations in hair than those in the single-housing system with no platform. Haptoglobin concentrations and kit mortality rates during lactation were greater for the collective-cage female rabbits. The collective group had more culled females and more lesions than in the other groups. The main reasons for culling in all the groups were reproduction problems and presence of abscesses, and the collective group of females was the most affected. In conclusion, it appears that keeping females together in collective systems negatively affects their health status and welfare, while single-housing systems imply lower kit mortality rates during lactation and cortisol concentrations, and fewer lesions in female rabbits.

Early deviations in performance, metabolic and immunological indicators affect stayability in rabbit females.

The main purpose of this study was to find several early factors affecting stayability in rabbit females. To reach this goal, 203 females were used from their first artificial insemination to their sixth parturition. Throughout that period, 48 traits were recorded, considered to be performance, metabolic and immunological indicators. These traits were initially recorded in females' first reproductive cycle. Later, removed females due to death or culling and those that were non-removed were identified. A first analysis was used to explore whether it was possible to classify females between those reaching and those not reaching up to the mean lifespan of a rabbit female (the fifth reproductive) cycle using information from the first reproductive cycle. The analysis results showed that 97% of the non-removed females were classified correctly, whereas only 60% of the removed females were classified as animals to be removed. The reason for this difference lies in the model's characteristics, which was designed using early traits and was able to classify only the cases in which females would be removed due to performance, metabolic or immunologic imbalances in their early lives. Our results suggest that the model defines the necessary conditions, but not the sufficient ones, for females to remain alive in the herd. The aim of a second analysis was to find out the main early differences between the non-removed and removed females. The live weights records taken in the first cycle indicated that the females removed in their first cycle were lighter, while those removed in their second cycle were heavier with longer stayability (-203 and +202 g on average, respectively; P < 0.05). Non-removed females showed higher glucose and lower beta-hydroxybutyrate concentrations in the first cycle than the removed females (+4.8 and -10.7%, respectively; P < 0.05). The average lymphocytes B counts in the first cycle were 22.7% higher in the non-removed females group (P < 0.05). The females removed in the first reproductive cycle presented a higher granulocytes/lymphocytes ratio in this cycle than those that at least reached the second cycle (4.81 v. 1.66; P < 0.001). Consequently, non-removed females at sixth parturition offered adequate body development and energy levels, less immunological stress and a more mature immune function in the first reproductive cycle. The females that deviated from this pattern were at higher risk of being removed from the herd.

Gene expression profiling of the SCN in young and old rhesus macaques.

In mammals, the suprachiasmatic nucleus (SCN) is the location of a master circadian pacemaker. It receives photic signals from the environment via the retinal hypothalamic tract, which play a key role in synchronizing the body's endogenously generated circadian rhythms with the 24-h rhythm of the environment. Therefore, it is plausible that age-related changes within the SCN contribute to the etiology of perturbed activity-rest cycles that become prevalent in humans during aging. To test this hypothesis, we used gene arrays and quantitative RT-PCR to profile age-related gene expression changes within the SCN of male rhesus macaques - a pragmatic translational animal model of human aging, which similarly displays an age-related attenuation of daytime activity levels. As expected, the SCN showed high expression of arginine vasopressin, vasoactive intestinal polypeptide, calbindin and nuclear receptor subfamily 1, group D, member 1 (NR1D1) (also known as reverse strand of ERBA (REV-ERBα), both at the mRNA and protein level. However, no obvious difference was detected between the SCNs of young (7-12 years) and old animals (21-26 years), in terms of the expression of core clock genes or genes associated with SCN signaling and neurotransmission. These data demonstrate the resilience of the primate SCN to normal aging, at least at the transcriptional level and, at least in males, suggest that age-related disruption of activity-rest cycles in humans may instead stem from changes within other components of the circadian system, such as desynchronization of subordinate oscillators in other parts of the body.

Long-term implications of feed energy source in different genetic types of reproductive rabbit females. II. Immunologic status.

Genetic selection and nutrition management have played a central role in the development of commercial rabbitry industry over the last few decades, being able to affect productive and immunological traits of the animals. However, the implication of different energy sources in animals from diverse genetic lines achieving such evolutionary success remains still unknown. Therefore, in this work, 203 female rabbits housed and bred in the same conditions were used from their first artificial insemination until their fifth weaning. The animals belonged to three different genetic types diverging greatly on breeding goals (H line, hyper-prolific (n=66); LP line, robust (n=67) and R line, selected for growth rate (n=67), and were assigned to two experimental diets, promoting major differences in energy source (cereal starch or animal fat)). The aims of this work were to: (1) characterize and describe blood leucocyte populations of three lines of rabbit does in different physiological stages during their reproductive period: first artificial insemination, first weaning, second parturition and fifth weaning; and (2) study the possible influence of two different experimental diets on the leucocyte populations in peripheral blood. Flow cytometry analyses were performed on blood samples taken from females at each different sampling stade. Lymphocyte populations at both weanings were characterized by significantly lower counts of total, CD5+ and CD8+ lymphocytes (-19.8, -21.7 and -44.6%; P<0.05), and higher counts of monocytes and granulocytes (+49.2 and +26.2%; P<0.05) than in the other stages. Females had higher blood counts of lymphocytes B, CD8+ and CD25+ and lower counts of CD4+ at first than at fifth weaning (+55.6, +85.8, +57.5, -14.5%; P<0.05). G/L ratio was higher at both weanings (P<0.05), and CD4+/CD8+ ratio increased progressively from the 1AI to the 5 W (P<0.001). Regarding the effect of genetic type in blood leucocyte counts, LP animals presented the highest counts for total, B, CD5+ and CD8+ lymphocytes (+16.7, +31.8, +24.5 and +38.7; P<0.05), but R rabbits showed the highest counts for monocytes and granulocytes (+25.3 and +27.6; P<0.05). The type of diet given during the reproductive life did not affect the leucocyte population counts. These results indicate that there are detectable variations in the leucocyte profile depending on the reproductive stage of the animal (parturition, weaning or none of them). Moreover, foundation for reproductive longevity criteria allows animals to be more capable of adapting to the challenges of the reproductive cycle from an immunological viewpoint.

Locomotor activity and the expression of orexin A and orexin B in aged female rhesus macaques.

Reduced activity has been linked to age-associated physiological changes but the underlying root cause is unclear. The goal of the present study was to compare the orexin neuronal system of old (23-29 years) female rhesus macaques with either active or sedentary 24-hour locomotor activity patterns. Using immunohistochemistry, we counted the number of orexin A and orexin B neurons in the lateral hypothalamic area of each animal. Overall, we observed no difference in the distribution pattern or number of either orexin A or orexin B immune-positive neurons between animals in the 2 groups. Thus, reduced activity in the elderly is unlikely to stem from a loss of orexin neuronal perikarya in the lateral hypothalamic area. This, however, does not rule out the possibility that the reduced activity stems from reduced orexin neuronal projections to arousal centers of the brain, such as the locus coeruleus, or from attenuated release of orexin.

Estimation of the invasive disease potential of Streptococcus pneumoniae in children by the use of direct capsular typing in clinical specimens.

Traditionally, invasiveness indexes have been based on culture methods. We aimed to establish a new classification of the invasive disease potential of pneumococcal serotypes causing invasive pediatric disease in the era of conjugate vaccines in Catalonia, Spain, by adding capsular typing of Streptococcus pneumoniae in direct sample. Two samples of children attended at the University Hospital Sant Joan de Déu (Barcelona, Spain) between 2007 and 2011 were compared: a first sample of 358 children with invasive pneumococcal disease and a second sample of 402 pneumococcal nasopharyngeal carriers selected from 714 healthy children admitted for minor surgical procedures. The most common invasive serotypes were 1 (20.1 %, n = 72), 19A (13.9 %, n = 50), 3 (12.3 %, n = 44), and 7FA (7.5 %, n = 27), whereas the most common serotypes in carriage were 19A (8.7 %, n = 38), 10FC33C (7.8 %, n = 34), 6C (6.9 %, n = 30), and 19FBC (5.5 %, n = 24). We detected a rate of cocolonization of 26.4 % (n = 89) among the 336 samples serotyped in the carriers population. Serotypes 1, 3, and 7FA were significantly associated with high invasiveness. Serotypes 6C, 10FC33C, 23A, 35B, 19FBC, 21, 11AD, 15BC, 23B, 34, and 6A were significantly associated with low invasiveness. Our results proved that the use of molecular techniques in direct sample for both the detection and the capsular identification of Streptococcus pneumoniae is very useful to obtain a more accurate calculation of the invasiveness of the different pneumococcal serotypes.

A robust rabbit line increases leucocyte counts at weaning and reduces mortality by digestive disorder during fattening.

The present work evaluates how a rabbit line selected for robustness and two other lines selected for productive criteria, could have affected the physiological maturity and blood leukocytes counts of young rabbits at weaning, as well as their possible effect on the subsequent performance and health status during the growing period. The study was conducted on a total of 2904 young rabbits weaned at 30 days, belonging to three different genetic types (line H, founded for litter size at birth and selected for litter size at weaning during 17 generations; line LP, characterised by robustness founded for reproductive longevity criteria and selected for litter size at weaning for 7 generations; and line R, founded and selected during 25 generations for average daily gain from the 4th to the 9th week of life). Two different diets were used during lactation. The two diets were both isoenergetic and isoprotein but their main energy source differed, being either animal fat (AF) or cereal starch (CS). Leucocyte subsets were characterised at weaning, and growing performance was studied until 58 days of age (feed intake, live weight, mortality by digestive disorders and morbidity) for both medicated and non-medicated dietary versions. At weaning, young rabbits fed an AF lactating diet evidenced greater B lymphocyte count (on av. +8.6 ± 3.5 × 10(6)/L; P < 0.05) than those fed a CS diet. With respect to H and R rabbits, blood from LP ones had higher counts for total (on av. 591 ± 167 × 10(6)/L; P < 0.05), B (on av. +11.05 ± 4.3 × 10(6)/L; P < 0.05), T CD5(+) (on av. +266 ± 83 × 10(6)/L; P < 0.05) and CD8(+) lymphocytes (on av. +72.5 ± 28 × 10(6)/L; P < 0.05), and with respect to R, higher counts of CD4(+) (on av. +121 ± 47 × 10(6)/L; P < 0.05) lymphocytes (on av. +12.3 ± 4.1 × 10(6)/L; P < 0.05), monocytes (on av. +66 ± 32 × 10(6)/L; P < 0.05) and granulocytes (on av. +567 ± 182 × 10(6)/L; P<0.05) at weaning. LP line rabbits also showed lower mortality by digestive disorders (on av. -8 points of percentage) and morbidity (on av. -4 points) than those from H and R lines during the growing period (P < 0.05). R animals presented higher feed intake and daily weight gain, and a lower feed conversion ratio than H and LP animals (on av. +16.7 ± 2.7 g dry matter/day, +10.3 ± 0.4 g/day and -0.22 ± 0.04 g dry matter/g, respectively). In conclusion, the foundation of a line for reproductive longevity, which has been previously reported to give greater robustness (low environmental sensitivity) to their reproductive stock, could have conferred higher leukocytes counts at weaning to their offspring, as well as a better ability to confront digestive disorders as compared to other lines founded or selected exclusively for productive criteria.

High prevalence of genetically-determined mannose binding lectin deficiency in young children with invasive pneumococcal disease.

Susceptibility to invasive pneumococcal disease (IPD) correlates with age, younger children being the group with the highest burden of disease. The relevance of the innate immune response and particularly the role of mannose-binding lectin (MBL) in combating IPD is not well known. This is a 2-year prospective study (February 2011 to March 2013) including patients with IPD who attended two hospitals from Catalonia, Spain. Variables including attack rate of pneumococcal serotype (high or low invasive potential serotypes) and genotypes associated with low serum MBL levels were recorded. One hundred and forty-seven patients were included in the study. One hundred and two (69.4%) patients were children or adolescents <18 years and 45 (30.6%) were adults. Overall, low-MBL genotypes (O/O; XA/O) were detected in 23 (15.6%) patients. Children <2 years showed a higher frequency of low-MBL genotypes compared with other patients (31.0% vs. 11.9%; p = 0.031). Further sub-analysis revealed a higher proportion of low-MBL genotypes in children <2 years with IPD caused by opportunistic or low-attack-rate serotypes when compared with older patients (46.2% vs. 13.2%; p = 0.02). However, no statistically significant differences between the two groups were observed when including patients infected with invasive or high-attack-rate serotypes (18.8% vs. 10.0%; p = 0.59). Our data suggest that young children with a genetically determined low-MBL production are at a higher risk of developing IPD, particularly that caused by opportunistic or low-attack-rate pneumococcal serotypes.

High invasiveness of pneumococcal serotypes included in the new generation of conjugate vaccines.

The implementation of the seven-valent pneumococcal conjugate vaccine, PCV7, has resulted in significant changes in the pneumococcal population being carried and causing disease. We aimed to determine the invasive disease potential of serotypes causing invasive paediatric disease in the era of conjugate vaccines in Catalonia, Spain, and their potential coverage by the 13-valent pneumococcal conjugate vaccine, PCV13. As a secondary objective, we evaluated whether implementation of PCV7 had resulted in significant changes in the invasive disease potential of the most frequent serotypes circulating in the area. Two pneumococcal collections obtained from children admitted to the University Hospital Sant Joan de Déu (Barcelona, Spain) between 2007 and 2011 were compared: a first set of 159 invasive disease isolates, and a second set of 209 nasopharyngeal isolates recovered from healthy children admitted for minor surgery. The most common invasive serotypes were 1 (24.5%, n = 39), 19A (21.2%, n = 34), 5 (8.8%, n = 14), 7F (8.8%, n = 14) and 3 (5%, n = 8). The most common serotypes in carriage were 19A (10%, n = 21), 6C (9%, n = 19), 23B (8.1%, n = 17), 6A (7.6%, n = 16) and 19F (6.2%, n = 13). A significantly higher propensity to cause invasive disease was observed for serotypes 1, 3, 5, 7F and 19A, all of which are included in PCV13. After false-discovery-rate correction, the results were robust for serotypes 1, 5, 7F and 19A. Non-PCV13 serotypes had a low invasive disease potential. Our data reinforce the need for continuous surveillance and should encourage efforts to introduce universal vaccination with PCV13 in children in our region.

Serotype 3 is a common serotype causing invasive pneumococcal disease in children less than 5 years old, as identified by real-time PCR.

Serotype 3 is one of the most often detected pneumococcal serotypes in adults and it is associated with serious disease. In contrast, the isolation of serotype 3 by bacterial culture is unusual in children with invasive pneumococcal disease (IPD). The purpose of this study was to learn the serotype distribution of IPD, including culture-negative episodes, by using molecular methods in normal sterile samples. We studied all children<5 years of age with IPD admitted to two paediatric hospitals in Catalonia, Spain, from 2007 to 2009. A sequential real-time polymerase chain reaction (PCR) approach was added to routine methods for the detection and serotyping of pneumococcal infection. Among 257 episodes (219 pneumonia, 27 meningitis, six bacteraemia and five others), 33.5% were identified by culture and the rest, 66.5%, were detected exclusively by real-time PCR. The most common serotypes detected by culture were serotypes 1 (26.7%) and 19A (25.6%), and by real-time PCR, serotypes 1 (19.8%) and 3 (18.1%). Theoretical coverage rates by the PCV7, PCV10 and PCV13 vaccines were 10.5, 52.3 and 87.2%, respectively, for those episodes identified by culture, compared to 5.3, 31.6 and 60.2% for those identified only by real-time PCR. Multiplex real-time PCR has been shown to be useful for surveillance studies of IPD. Serotype 3 is underdiagnosed by culture and is important in paediatric IPD.

Low prevalence of invasive bacterial infection in febrile infants under 3 months of age with enterovirus infection.

Infants under 3 months of age with fever without source (FWS) generally undergo a full, invasive septic evaluation to exclude invasive bacterial infection (IBI). Enterovirus (EV) infections are mostly banal and self-limiting and show a high prevalence rate at this age. We aimed to investigate the prevalence of IBI in EV-infected and uninfected infants under 3 months of age with FWS. This was a prospective observational cohort study of infants aged <90 days who were admitted because of FWS. As per protocol, blood and urine analysis and culture were obtained in all cases, and RNA EV from blood and/or cerebrospinal fluid samples was determined by real-time PCR. Three hundred and eighty-one previously healthy infants with FWS were included. EV infection was diagnosed in 64 children (16.8%; 95% confidence interval, 13.2-20.9%) and showed an uneventful evolution in all cases. Laboratory markers of infection were consistently lower in EV-infected patients; only one case of IBI (1.6%) was observed in an EV-infected patient as compared with 25.2% in EV-negative infants (p <0.001). Intravenous antibiotic use and length of stay were no different in EV-infected and uninfected patients. In our study, febrile infants (<90 days) diagnosed with EV infection showed a low risk of IBI when compared with uninfected patients. The systematic investigation of EV infection in young infants with FWS may allow a more conservative approach to the management of these patients. Further studies on this diagnostic approach are needed.

Comparison of an in-house real-time RT-PCR assay with a commercial assay for detection of enterovirus RNA in clinical samples.

Molecular detection of enterovirus (EV) RNA based on PCR methods is a quicker and more sensitive approach than culture methods. At present, different PCR-based methods for EV RNA detection are available, but comparisons of results obtained according to the different approaches are limited. We evaluated an in-house real-time RT-PCR assay with a commercialized TaqMan real-time RT-PCR kit for detection of EV. Consecutive clinical specimens from paediatric patients less than 6 years old with clinical suspicion of EV infection were analyzed between July and November 2010. After RNA extraction, samples were amplified both by the real-time RT-PCR commercial assay and the in-house assay. A total of 19 of 132 patients (14.4%) involving 20 samples (14 plasma samples and 6 CSF) were positive in at least one of the two assays. The sensitivity of the in-house assay when the MutaPLATE® assay was used as a reference was 90% (IC 95%; 74.35-100) and the specificity was 100% (IC 95%; 99.63-100). Cts results of two methods were statistically correlated (r = 0.774; P = 0.01). In conclusion, these two real-time RT-PCR assays are rapid and easy methods for detection of EV.

Streptococcus pneumoniae serotype 1 causing invasive disease among children in Barcelona over a 20-year period (1989-2008).

Fifty-six isolates of serotype 1 were identified during a 20-year prospective study (1989-2008), including all children with culture-proven invasive pneumococcal disease (IPD) admitted to a children's hospital in Barcelona. Forty-eight of them (85.7%) were in children aged >2 years. Complicated pneumonia (n = 28) and non-complicated pneumonia (n = 20) were the main clinical presentations. The frequency of serotype 1 IPD increased from 1999-2003 to 2004-2008: 1.2 to 4.4 episodes/100 000 children (p <0.001). The ST306 clone were identified in 70.4% of isolates. As IPD caused by serotype 1 is mainly detected in older children, a vaccination programme for children >2 years should be considered.

Strains of Staphylococcus aureus and pathology associated with chronic suppurative mastitis in rabbits.

Staphylococcal mastitis is one of the main reasons for culling adult does from commercial rabbitries. The aim of this study was describe the spectrum of gross and microscopic lesions in 178 cases of chronic staphylococcal mastitis in adult does and to determine whether there is a correlation between Staphylococcus aureus genotypes and pathology. On the basis of histopathology, chronic mastitis was differentiated into abscesses (66.3%), suppurative mastitis with a lobular pattern (7.9%), cellulitis (19.6%) and mixed lesions (6.2%). Pathological presentations were not related to S. aureus genotype.

Predictive factors for the detection of CIN II-III in the follow-up of women with CIN I.

PURPOSE OF INVESTIGATION: To determine which factors may increase the risk that women diagnosed with CIN I may later develop CIN II-III. METHODS: A prospective study of 174 women with a grade 1 intraepithelial lesion (CIN I) confirmed by biopsy, with a follow-up time of at least one year. The following factors were studied: age, HPV infection, HPV infection by a high-risk genotype, the HPV genotypes involved, coinfection by several HPV genotypes and duration of follow-up. These factors were correlated with later detection of CIN II-III by biopsy during follow-up. Statistical analysis was performed using SPSS. RESULTS: CIN II-III was detected at the follow-up in 24 of 174 women included in the study (13.7%), in four cases by colposcopically directed biopsy and in 20 by LLETZ. Correlation of the factors studied with the incidence of CIN II-III in this group showed that the only statistically significant factors were overall HPV infection and HPV infection by genotypes 31 and 70 (Chi-square and Fisher's test, p < 0.05, respectively), while the duration of follow-up came close to statistical significance (Student's test, p = 0.052). CONCLUSION: HPV infection and duration of follow-up are predictive factors for the detection of CIN II-III in follow-up care for women with CIN I.

Staphylococcal infections in rabbit does on two industrial farms.

The main reasons for culling adult rabbit does on two Spanish rabbit farms were investigated for a year. The most important conditions were mastitis (33.3 per cent), followed by subcutaneous abscesses (9.9 per cent) and pyometra (8.7 per cent). Staphylococcus aureus infections were the most severe problem, the organism being isolated from 69.2 per cent of infected animals. Pasteurella species were more prevalent in cases of pyometra and pneumonia. Two strains of S aureus were identified by using polymorphism of the coagulase gene as the criterion. One of these strains was responsible for the majority of the staphylococcal infections and was isolated from several pathological processes.