Fluoroquinolone and beta-lactam antimicrobials induce different transcriptome profiles in Salmonella enterica persister cells.

Here, we investigate the transcriptome profiles of two S. Enteritidis and one S. Schwarzengrund isolates that present different persister levels when exposed to ciprofloxacin or ceftazidime. It was possible to note a distinct transcript profile among isolates, time of exposure, and treatment. We could not find a commonly expressed transcript profile that plays a role in persister formation after S. enterica exposure to beta-lactam or fluoroquinolone, as only three DEGs presented the same behavior under the conditions and isolates tested. It appears that the formation of persisters in S. enterica after exposure to ciprofloxacin is linked to the overexpression of genes involved in the SOS response (recA), cell division inhibitor (sulA), iron-sulfur metabolism (hscA and iscS), and type I TA system (tisB). On the other hand, most genes differentially expressed in S. enterica after exposure to ceftazidime appeared to be downregulated and were part of the flagellar assembly apparatus, citrate cycle (TCA cycle), glycolysis/gluconeogenesis, carbon metabolism, bacterial secretion system, quorum sensing, pyruvate metabolism pathway, and biosynthesis of secondary metabolites. The different transcriptome profiles found in S. enterica persisters induced by ciprofloxacin and ceftazidime suggest that these cells modulate their response differently according to each stress.

Effects of natural aging and gender on pro-inflammatory markers.

The term inflammaging is now widely used to designate the inflammatory process of natural aging. During this process, cytokine balance is altered, presumably due to the loss of homeostasis, thus contributing to a greater predisposition to disease and exacerbation of chronic diseases. The aim of the study was to analyze the relationship between pro-inflammatory markers and age in the natural aging process of healthy individuals. One hundred and ten subjects were divided into 5 groups according to age (22 subjects/group). Interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α) were quantified using the ELISA method. High-sensitivity C-reactive protein (hsCRP) was analyzed by turbidimetry according to laboratory procedures. The main findings of this study were: a positive correlation between hsCRP and IL-6 as a function of age (110 subjects); women showed stronger correlations; the 51-60 age group had the highest values for hsCRP and IL-6; women presented higher values for hsCRP in the 51-60 age group and higher values for IL-6 in the 61-70 age group; and men showed higher values in the 51-60 age group for hsCRP and IL-6. In conclusion, the natural aging process increased IL-6 and hsCRP levels, which is consistent with the inflammaging theory; however, women presented stronger correlations compared to men (IL-6 and hsCRP) and the 51-60 age range seems to be a key point for these increases. These findings are important because they indicate that early preventive measures may minimize the increase in these inflammatory markers in natural human aging.

Effects of natural aging and gender on pro-inflammatory markers

The term inflammaging is now widely used to designate the inflammatory process of natural aging. During this process, cytokine balance is altered, presumably due to the loss of homeostasis, thus contributing to a greater predisposition to disease and exacerbation of chronic diseases. The aim of the study was to analyze the relationship between pro-inflammatory markers and age in the natural aging process of healthy individuals. One hundred and ten subjects were divided into 5 groups according to age (22 subjects/group). Interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α) were quantified using the ELISA method. High-sensitivity C-reactive protein (hsCRP) was analyzed by turbidimetry according to laboratory procedures. The main findings of this study were: a positive correlation between hsCRP and IL-6 as a function of age (110 subjects); women showed stronger correlations; the 51-60 age group had the highest values for hsCRP and IL-6; women presented higher values for hsCRP in the 51-60 age group and higher values for IL-6 in the 61-70 age group; and men showed higher values in the 51-60 age group for hsCRP and IL-6. In conclusion, the natural aging process increased IL-6 and hsCRP levels, which is consistent with the inflammaging theory; however, women presented stronger correlations compared to men (IL-6 and hsCRP) and the 51-60 age range seems to be a key point for these increases. These findings are important because they indicate that early preventive measures may minimize the increase in these inflammatory markers in natural human aging.

Inter-observer reliability of animal-based welfare indicators included in the Animal Welfare Indicators welfare assessment protocol for dairy goats.

This study was conducted within the context of the Animal Welfare Indicators (AWIN) project and the underlying scientific motivation for the development of the study was the scarcity of data regarding inter-observer reliability (IOR) of welfare indicators, particularly given the importance of reliability as a further step for developing on-farm welfare assessment protocols. The objective of this study is therefore to evaluate IOR of animal-based indicators (at group and individual-level) of the AWIN welfare assessment protocol (prototype) for dairy goats. In the design of the study, two pairs of observers, one in Portugal and another in Italy, visited 10 farms each and applied the AWIN prototype protocol. Farms in both countries were visited between January and March 2014, and all the observers received the same training before the farm visits were initiated. Data collected during farm visits, and analysed in this study, include group-level and individual-level observations. The results of our study allow us to conclude that most of the group-level indicators presented the highest IOR level ('substantial', 0.85 to 0.99) in both field studies, pointing to a usable set of animal-based welfare indicators that were therefore included in the first level of the final AWIN welfare assessment protocol for dairy goats. Inter-observer reliability of individual-level indicators was lower, but the majority of them still reached 'fair to good' (0.41 to 0.75) and 'excellent' (0.76 to 1) levels. In the paper we explore reasons for the differences found in IOR between the group and individual-level indicators, including how the number of individual-level indicators to be assessed on each animal and the restraining method may have affected the results. Furthermore, we discuss the differences found in the IOR of individual-level indicators in both countries: the Portuguese pair of observers reached a higher level of IOR, when compared with the Italian observers. We argue how the reasons behind these differences may stem from the restraining method applied, or the different background and experience of the observers. Finally, the discussion of the results emphasizes the importance of considering that reliability is not an absolute attribute of an indicator, but derives from an interaction between the indicators, the observers and the situation in which the assessment is taking place. This highlights the importance of further considering the indicators' reliability while developing welfare assessment protocols.

Consistency over time of animal-based welfare indicators as a further step for developing a welfare assessment monitoring scheme: The case of the Animal Welfare Indicators protocol for dairy goats.

Consistency over time (COT) of animal-based indicators is key to a reliable and feasible welfare protocol, indicating that results are representative over long-term situations. High levels of consistency ensure fairness for the farmer and credibility of the system. In addition, indicator COT reduces recording costs, as having indicators that do not change over a long period of time will require less farm visits to achieve reliable estimates. To date, COT of animal-based indicators included in the welfare assessment of dairy goats has never been tested. Therefore, our aim was to investigate COT of animal-based indicators included in the Animal Welfare Indicators (AWIN) welfare assessment prototype protocol for dairy goats. To meet this goal, a study was designed where an average of 3 mo elapsed between 2 sets of visits to the same 20 dairy goat farms (10 in Portugal and 10 in Italy), with no major changes in management routines or housing conditions occurring during this period. Initially, we performed a Wilcoxon signed rank test to investigate whether the results obtained during the 2 visits were significantly different. After this preliminary screening, the indicators presenting nonsignificant differences between visits were submitted to a second step analysis, where discriminative and evaluative analyses were conducted to reach a final indicator lineup. The discriminative approach helped distinguishing among farms, whereas the agreement analysis showed us the range of differences between repeated assessments. Some particular conclusions could be drawn from this combined analysis, helping to the development of the final AWIN welfare assessment protocol for dairy goats and as a further step to develop a welfare assessment monitoring scheme for this and other species. In this sense, the AWIN welfare assessment protocol allows for the quick differentiation between farms based on the identification of persistent welfare problems, by recording highly consistent and feasible indicators. In a second step, a more comprehensive protocol, consisting of indicators more likely to be subject to variations along time, was applied. Repeated assessments and long-term studies of indicator consistency are needed to help determine the frequency of visits required to obtain a consistent and feasible welfare assessment scheme. This paper adds to the literature by providing guidance on the variability of animal-based indicators over time.

Measuring postural control during mini-squat posture in men with early knee osteoarthritis.

Studies have suggested a compromised postural control in individuals with knee osteoarthritis (OA) evidenced by larger and faster displacement of center of pressure (COP). However, quantification of postural control in the mini-squat posture performed by patients with early knee OA and its relation to muscle strength and self-reported symptoms have not been investigated. The main aim of this cross-sectional, observational, controlled study was to determine whether postural control in the mini-squat posture differs between individuals with early knee OA and a control group (CG) and verify the relation among knee extensor torque (KET) and self-reported physical function, stiffness and pain. Twenty four individuals with knee OA grades I and II (OAG) (mean age: 52.35±5.00) and twenty subjects without knee injuries (CG) (mean age: 51.40±8.07) participated in this study. Participants were assessed in postural control through a force plate (Bertec Mod. USA), which provided information about the anterior-posterior (AP) and medial-lateral (ML) COP displacement during the mini-squat, in isometric, concentric and eccentric knee extensor torque (KET) (90°/s) through an isokinetic dynamometer (BiodexMulti-Joint System3, Biodex Medical Incorporation, New York, NY, USA), and in self-reported symptoms through the WOMAC questionnaire. The main outcomes measured were the AP and ML COP amplitude and velocity of displacement; isometric, concentric, and eccentric KET and self-reported physical function, stiffness and pain. No significant differences were found between groups for postural control (p>0.05). Significant lower eccentric KET (p=0.01) and higher scores for the WOMAC subscales of pain (p=<0.001), stiffness (p=0.001) and physical function (p<0.001) were found for the OAG. Moderate and negative correlations were found between the AP COP amplitude of displacement and physical function (ρ=-0.40, p=0.02). Moderate and negative correlations were observed between the AP COP velocity of displacement and physical function (ρ=0.47, p=0.01) and stiffness (ρ=-0.45, p=0.02). The findings of the present study emphasize the importance of rehabilitation from the early degrees of knee OA to prevent postural instability and the need to include quadriceps muscle strengthening, especially by eccentric contractions. The relationship between the self-reported symptoms and a lower and slower COP displacement suggest that the postural control strategy during tasks with a semi-flexed knee should be further investigated.

Invited review: Animal-based indicators for on-farm welfare assessment for dairy goats.

This paper reviews animal-based welfare indicators to develop a valid, reliable, and feasible on-farm welfare assessment protocol for dairy goats. The indicators were considered in the light of the 4 accepted principles (good feeding, good housing, good health, appropriate behavior) subdivided into 12 criteria developed by the European Welfare Quality program. We will only examine the practical indicators to be used on-farm, excluding those requiring the use of specific instruments or laboratory analysis and those that are recorded at the slaughterhouse. Body condition score, hair coat condition, and queuing at the feed barrier or at the drinker seem the most promising indicators for the assessment of the "good feeding" principle. As to "good housing," some indicators were considered promising for assessing "comfort around resting" (e.g., resting in contact with a wall) or "thermal comfort" (e.g., panting score for the detection of heat stress and shivering score for the detection of cold stress). Several indicators related to "good health," such as lameness, claw overgrowth, presence of external abscesses, and hair coat condition, were identified. As to the "appropriate behavior" principle, different criteria have been identified: agonistic behavior is largely used as the "expression of social behavior" criterion, but it is often not feasible for on-farm assessment. Latency to first contact and the avoidance distance test can be used as criteria for assessing the quality of the human-animal relationship. Qualitative behavior assessment seems to be a promising indicator for addressing the "positive emotional state" criterion. Promising indicators were identified for most of the considered criteria; however, no valid indicator has been identified for "expression of other behaviors." Interobserver reliability has rarely been assessed and warrants further attention; in contrast, short-term intraobserver reliability is frequently assessed and some studies consider mid- and long-term reliability. The feasibility of most of the reviewed indicators in commercial farms still needs to be carefully evaluated, as several studies were performed under experimental conditions. Our review highlights some aspects of goat welfare that have been widely studied, but some indicators need to be investigated further and drafted before being included in a valid, reliable, and feasible welfare assessment protocol. The indicators selected and examined may be an invaluable starting point for the development of an on-farm welfare assessment protocol for dairy goats.

Enterohepatic Helicobacter spp. in colonic biopsies of dogs: molecular, histopathological and immunohistochemical investigations.

Enterohepatic Helicobacter spp. have been described colonizing the large intestine and liver of healthy and symptomatic subjects and are thought to have a role in the development of inflammatory bowel disease (IBD). The prevalence of enterohepatic Helicobacter spp. infection in dogs is largely unknown and to our knowledge there are no data about their potential pathogenic role. In light of these considerations, the aims of this study were (i) to assess the prevalence of enterohepatic Helicobacter spp. in colonic biopsies of symptomatic pet dogs and (ii) to evaluate a possible association between Helicobacter spp. colonization status (heavily colonized, poorly colonized and uncolonized biopsies) and histological lesions. Colonic biopsies from 27 pet dogs of different ages were evaluated by family Helicobacteraceae and enterohepatic Helicobacter spp. PCR, histology, and immunohistochemistry for the in situ detection of Helicobacter spp. organisms. 85% and 52% of colonic biopsies were positive by Helicobacteraceae and enterohepatic Helicobacter spp. PCR, respectively. Immunohistochemistry revealed Helicobacter spp. were localized both in the superficial mucus (55%) and within intestinal crypts (33%). Dogs with heavy enterohepatic Helicobacter spp. colonization were significantly younger and had a higher level of mucosal fibrosis/atrophy than dogs with uncolonized or poorly colonized biopsies (p<0.05). These findings contribute to widen current knowledge regarding canine enterohepatic Helicobacter spp., suggesting the infection is rather common in dogs and acquired at an early age. Furthermore, heavy colonization of colonic crypts is associated with chronic inflammatory lesions (fibrosis/atrophy), supporting the role of enterohepatic Helicobacter spp. in the development of canine IBD.

Short communication: Breed differences affecting dairy cattle welfare in traditional alpine tie-stall husbandry systems.

The aim of this investigation was to compare the prevalence of indicators of poor welfare among 5 Italian cattle breeds (Italian Holstein-Friesian, Italian Bruna, Pezzata Rossa Italiana, Grigia Alpina, and Pezzata Rossa d'Oropa) kept in tie-stalls in the Italian Alps under similar housing and management conditions. We recorded the presence of integument alterations (hairless patch areas, lesion/swollen areas, or overgrown claws) and lameness in 612 cows. Additionally, we checked 834 cows for the presence of physical malformations ("open" shoulders). In general, the prevalence of welfare problems showed a decreasing trend from the more productive to the less productive breeds. Local breeds (Grigia Alpina and Pezzata Rossa d'Oropa) showed a significantly lower prevalence of welfare problems compared with the other 3 breeds, whereas Italian Holstein-Friesian usually had the highest percentage of individuals with problems. No differences were found between Pezzata Rossa Italiana and Italian Bruna, both of which showed fewer problems than Italian Holstein-Friesian. The effect of the breed significantly affected the welfare of dairy cows in tie-stalls in alpine traditional husbandry systems. The prevalence of the negative welfare indicators studied was lower in local breeds, which are better adapted to local breeding conditions. Our results indicate an urgent need to promote changes in the criteria used for genetic selection in the dairy industry and underline the importance of maintaining the diversity of local breeds, which should be carefully chosen for each specific environmental condition.

Mammary tumor phenotypes in wild-type aging female FVB/N mice with pituitary prolactinomas.

Prolactin-secreting pituitary adenomas are common spontaneous lesions in aging FVB females. Prolactin-secreting pituitary proliferations play a significant role in mouse mammary tumorigenesis generally producing adenosquamous carcinomas. Since genetically engineered FVB mice are frequently used to study mammary tumor biology, we have examined a cohort of 64 aging wild-type FVB/N females to establish the prevalence and the nature of spontaneous mammary and pituitary tumors. Tissues from mammary and pituitary glands were studied by histopathology and immunohistochemistry. Of the 64 examined mice, 20 had pituitary tumors and 20 had mammary tumors. Mammary and pituitary tumors were associated in 17 mice. All pituitary tumors were prolactin-positive by immunohistochemistry and classified as prolactinomas. Fourteen mammary tumors, including 12 cases with and 2 without concurrent prolactinomas, were adenocarcinomas with different combinations of epithelial growth patterns. Five mice with prolactinomas had mammary tumors characterized by the epithelial-mesenchymal transition (EMT) phenotype. Estrogen receptor alpha (ERalpha)-positivity was observed for 14 of the 18 mammary tumors tested, including both adenocarcinomas with nuclear immunoreactivity and EMT-phenotype tumors with both nuclear and cytoplasmic immunoreactivity. No immunoreactivity for the progesterone receptor was observed. This study confirms that spontaneous prolactinomas and mammary tumors are both common and significantly associated lesions in FVB mice. Parity and age represented risk factors for the development of these tumors. Compared with previous reports, prolactinoma-associated mammary tumors displayed a broader morphologic spectrum, including cases with the EMT phenotype. The elevated number of prolactinoma-associated and ERalpha-positive mammary tumors opens intriguing possibilities concerning the role of ERalpha cytoplasmic localization during EMT tumorigenesis.

Expression of major histocompatibility complex class II antigens in porcine leptospiral nephritis.

Class II major histocompatibility complex (MHCII) is required for the presentation of antigens to CD4 helper T cells. During nephritis, not only primary antigen presenting cells such as histiocytes and lymphocytes, but also cytokine-stimulated tubular epithelial cells express MHCII. Leptospirosis in fattening pigs is characterized by several degrees of nephritis, from absence of lesions to severe multifocal tubulo-interstitial inflammation. Renal tissue from 20 8-month-old pigs with spontaneous nephritis and 6 control pigs without renal lesions were investigated for leptospirosis by indirect immunohistochemistry (IHC) and polymerase chain reaction (PCR). IHC for MHCII also was performed on renal samples. Serum samples were tested for different serovars of Leptospira interrogans. Control pigs were free of interstitial nephritis and negative for leptospirosis by all tests. In pigs with nephritis, serology was positive for serovar Pomona in 19/20 pigs. In 16 of these 19 pigs, leptospiral renal infection was confirmed by PCR and/or indirect IHC. Nephritic lesions were classified histologically into perivascular lymphocytic (4 pigs), lymphofollicular (6 pigs), lymphohistiocytic (8 pigs), and neutrophilic (2 pigs) pattern. MHCII expression by histiocytes and lymphocytes was observed in all lesions. Prominent MHCII expression in regenerating tubular epithelium was observed in lymphofollicular and lymphohistiocytic nephritis. No tubular colocalization between leptospiral and MHCII antigen was observed. Results suggest that during leptospiral nephritis, MHCII contributes to the intensity of the inflammatory response. Furthermore de novo MHCII expression in regenerating tubules may play a role in the defence mechanism against leptospiral tubular colonization.

The provision of drinking water to veal calves for welfare purposes.

Growth performance, behavior, physiology, forestomach development, abomasal lesions, and meat quality of veal calves fed a milk-replacer diet (No Water) were compared to those obtained from calves fed the same diet and provided with increasing amounts of drinking water (Water). Two groups of 69 Polish Friesian calves, balanced according to initial BW, were assigned to two water treatments in a 3 x 2 x 2 factorial arrangement that provided solid feed in addition to the milk-replacer diet (No solid feed, 250 g x calf(-1) x d(-1) of wheat straw or the same amount of beet pulp), and the adoption of two housing systems (individual stall vs group pen). The fattening trial lasted 160 d, and calves received drinking water starting from the 2nd wk of the study. The amount of drinking water was progressively increased from 3 to 8 L x cal(-1) x d(-1). Although not dehydrated, as shown by hematocrit and Na, K, and total protein hemoconcentration, calves consumed almost all the offered amount of water throughout the fattening period. Therefore, the water provided by the milk replacer alone, which ranged between 6 to 16 L calf d(-1), was not sufficient to satisfy the need of the animal. Drinking water did not affect the calves' growth performance but it reduced nonnutritive oral behavior throughout the fattening period. Based on these results, drinking water did not cover a shortage in the calves' water requirement but it played a role in environmental enrichment. Health status was similar between treatments, although water provision reduced the episodes of feed refusal. The measurement of chronic stress by ACTH challenge showed that the administration of drinking water would be advisable when calves are fed with small amounts of solid feed for well-being. Feces consistency and animal cleanliness were not affected by drinking water. At slaughter, forestomach development was similar between treatments, and drinking water did not affect the number of calves showing rumen hair-balls and abomasal lesions. No differences in color and other meat quality traits were observed between Water and No Water calves. Despite the lack of direct effects on productive traits, when water was available, the calves drank it, and positive effects were noticed on their nonnutritive oral behaviors and chronic stress indicators.

The provision of solid feeds to veal calves: II. Behavior, physiology, and abomasal damage.

The aim of this study was to evaluate the effects of the addition of two roughage sources (wheat straw and beet pulp) to the milk replacer diet of veal calves, in order to reduce stress and improve animal welfare. We allocated 138 Polish Friesian male calves to three different feeding plans: a milk replacer diet (Control), 250 g/d of wheat straw in addition to the milk replacer, or 250 g/d of dried beet pulp in addition to the milk replacer. Within each feeding treatment, 16 calves were individually housed and 30 were kept in group pens (five calves/pen). Several behavioral, physiological, and health welfare indicators were monitored throughout the fattening period, which lasted for 160 d. Abnormal oral behavior around the meals was higher in Control calves (P < 0.01), while its lowest level was observed in straw-fed calves. At the beginning of the trial, chewing was higher in calves receiving solid feeds (P < 0.001), but the difference from the Control gradually decreased and disappeared at wk 13 for calves fed beet pulp and at wk 17 for those fed wheat straw. At the end of the fattening period, no differences among treatments were found in the frequency of chewing. Regardless of the diet, self-grooming decreased with age and no relationship was observed between this behavior and the presence of rumen hairballs. Cross-sucking was performed with low frequencies (from 4.70% at wk 2 to 1.05% at wk 23 around the meals, and even lower far from the meals) and was not affected by the provision of roughage. The time in contact with the bucket during the whole day was higher in Controls, whereas calves fed wheat straw maintained a lower level of this activity until the end of the trial (P < 0.01). The calves fed wheat straw spent more time in contact with the feed trough (P < 0.001) than those fed beet pulp and Control calves. No differences were found in cortisol curves due to the feeding treatment. In calves fed beet pulp, most hematological measures statistically differed from the other treatments, possibly in response to the higher iron intake and(or) to the higher hemoconcentration, probably due to the administration of beet pulp as dried feed. The incidence of abomasal ulcers and erosions was increased by the provision of the solid feeds, particularly by a structured fiber source such as straw. A roughage source able to satisfy calves' behavioral needs and to improve digestive processes without damaging the digestive apparatus still has to be identified.

The provision of solid feeds to veal calves: I. Growth performance, forestomach development, and carcass and meat quality.

Growth performance, forestomach development, and carcass and meat quality of veal calves fed a milk replacer diet (Control) were compared to those obtained from calves fed the same liquid diet plus 250 g x calf(-1) x d(-1) of dried beet pulp or wheat straw. Three groups of 46 Polish Friesian calves, balanced according to initial BW, were assigned to the three dietary treatments in a fattening trial, which lasted 160 d. The provision of either solid feed did not affect the milk replacer intake. However, calves' ADG was increased (P < 0.01) only by feeding the beet pulp diet. The administration of both solid feeds improved calves' health status; calves fed solid feeds required fewer iron treatments for low hemoglobin and needed less medical treatments for respiratory or gastrointestinal diseases. In comparison to the Control calves, the provision of wheat straw and beet pulp increased iron intake throughout the fattening period by 41 and 130%, respectively. However, only calves fed beet pulp showed higher levels of hemoglobin and plasma iron concentrations (P < 0.05), whereas the same blood parameters were similar between Control calves and those fed wheat straw. At slaughter, both solid feeds led to empty forestomach weights heavier than those of Controls without reducing dressing percentage. The reticulorumen was heaviest in calves fed beet pulp, whereas wheat straw promoted omasal development. The administration of beet pulp resulted in a better carcass conformation than did the Control diet or wheat straw, but it had a detrimental effect on carcass color, which was graded as the darkest (P < 0.001). Consistent with this result, meat color of calves fed beet pulp was darker than that of Control calves and those fed wheat straw, because of the higher hematin concentration measured at the muscle level. No differences in carcass and meat color were observed between Control calves and calves fed wheat straw. The administration of solid feeds for welfare purposes does not always prevent the production of veal meat fulfilling the color standards required by the market. There is not a straight-forward relationship between a solid feed's iron content and the "redness" of veal meat, which should be related to the capability of the calves to use the iron provided by the roughage.

Spironolactone as a single agent for long-term therapy of hirsute patients.

OBJECTIVE: To assess the androgen-suppressing effect of spironolactone, and the use of this drug as a single agent in the long-term therapy of hirsute patients with either polycystic ovary syndrome (PCOS) or idiopathic hirsutism (IH). Standard cyproterone acetate (CPA) treatment was used to evaluate the results obtained with spironolactone. DESIGN: Prospective randomized study. PATIENTS: Forty-six hirsute women were separated randomly into two groups, stratified for polycystic ovary syndrome. For 12 months, Group 1 (21 patients, 10 PCOS) received spironolactone only (200 mg/day). Group 2 (23 patients, nine PCOS) received CPA (50 mg/day) with ethinyl oestradiol (35 microgram/day). MEASUREMENTS: Ferriman-Gallwey clinical score for hirsutism and serum testosterone, androstenedione, and LH levels. RESULTS: In IH patients, hirsutism regressed equally with spironolactone (21 +/- 2-14.5 +/- 2) and CPA (23 +/- 2-13 +/- 2). In PCOS patients, the mean score for hirsutism after 12 months was significantly lower with CPA (12 +/- 1) than with spironolactone (16 +/- 1). Testosterone levels did not change with spironolactone; with CPA there was a decrease from baseline in PCOS (47% and 51%, 6 and 12 months) and IH patients (31% and 30%). Androstenedione levels also declined from baseline in CPA-treated PCOS patients (38% and 39%, 6 and 12 months). Androgen levels were significantly different between the groups after 6 and 12 months. LH levels decreased with CPA (72%) but not with spironolactone. CONCLUSION: Our results suggest that spironolactone used as a single agent is as effective as cyproterone acetate combined with oestradiol for long-term treatment of patients with idiopathic hirsutism. In PCOS patients, spironolactone is still effective for reducing hirsutism; however, for treatment of the hormonal or metabolic manifestations associated with PCOS, it may be necessary to combine spironolactone with either an antigonadotrophic agent or a drug that improves peripheral insulin sensitivity.

Cilia-associated respiratory (CAR) bacillus infection in conventionally reared rabbits.

This study was designed to investigate the prevalence of Cilia-associated respiratory (CAR) bacillus infection in rabbits reared for meat production in Italy and to correlate the presence of CAR bacillus with inflammatory lesions of the respiratory tract. Seventy health, 3-month-old, New Zealand White rabbits, raised in 10 different rabbitries in Northern Italy were randomly selected at slaughter. No gross lesions were found at necropsy in any rabbit. In each animal, the trachea and lungs were sampled, fixed in 10% formalin, embedded in paraffin and stained with the Warthin-Starry method to evaluate the presence of CAR bacillus, and with haematoxylin and eosin to evaluate the presence of inflammatory lesions. CAR bacillus was present in 50 out of 70 rabbits (71.4%) with a prevalence of the infection that varied from 30% to 100% in the seven rabbitries. CAR bacillus was present both in the trachea and bronchi in 23 cases (32.8%), only in the trachea in 24 cases (34.3%) and only in the bronchi in three cases (4.3%). Inflammatory lesions were found in the trachea (22 cases, 31.4%) and the bronchi (58 cases, 82.8). There was a strong, statically significant correlation between the presence of CAR bacillus in the bronchi and bronchial inflammatory lesions (P < 0.0001). This study indicates that CAR bacillus infection is widespread in conventionally reared rabbits in Italy and that a possible correlation exists between the presence of CAR bacillus and bronchial inflammatory lesions.

Feeding and social behavior of fallow deer (Dama dama L.) under intensive pasture confinement.

A population of 110 adult fallow deer kept in an enclosure under very intensive conditions (31.4 deer/ha) was observed for 12 h/d (from 0800 to 2000) for 17 d during the birth season in order to study social and feeding behavior under intensive management. Observations were carried out by a scan sampling technique. The feeding activity of all the deer that were not hidden behind trees or other obstacles in the enclosure (60 to 90% of all deer) and the composition and position of all groups were recorded every 20 min by video camera. Adult females ate more pasture than adult males, whereas adult males ate more hay and meal than females (P < .001). Feeding activity was concentrated in the morning and in the evening, but it was higher for bucks in the morning (when supplementary feed was distributed) than in the midday and in the evening (P < .001). The area where hay and meal were supplied was more frequently attended in morning by small groups of males. Group size was affected by the time of day (P < .05), being larger in the evening (13.6 +/- 1.2 in the evening vs 9.8 +/- 1.0 in the morning and 11.9 +/- 0.9 in the afternoon), when groups of females with fawns and mixed-sex groups (including at least one male) gathered into the pasture areas for grazing. Our study suggests that even under very intensive conditions fallow deer may tend to maintain the sexual segregation typical of this species during the birth season, but mixed-sex groups are still the most common type of social aggregation. This situation was particularly evident in the areas and at the time in which feeding activity was higher and might indicate that feeding requirements are more important than social needs when the space available is limited.