Male ruff colour as a rank signal in a monomorphic-horned mammal: behavioural correlates.

Coexistence of individuals within a social group is possible through the establishment of a hierarchy. Social dominance is achieved through aggressive interactions, and, in wild sheep and goats, it is related mainly to age, body size and weapon size as rank signals. Adult male Himalayan tahr are much larger than females and subadult males. They have a prominent neck ruff, ranging in colour from yellow (5.5-9.5 years old, i.e. young adults, golden males) to brown (7.5-14.5 years old, i.e. older individuals, pale and dark brown males), with golden males being the most dominant. We investigated the social behaviour of male tahr and analysed the relationships between ruff colour, courtship and agonistic behaviour patterns during the rut. Colour classes varied in their use of several behaviour patterns (male dominance: approach, stare, horning vegetation; courtship: low stretch, naso-genital contact, rush). Golden-ruffed males used more threats than darker ones. Pale brown and dark brown males addressed threats significantly more often to males of lower or their own colour classes, respectively, whereas golden ones addressed threats to all colour classes, including their own. The courtship of dominant males was characterised by the assertive rush, whereas that of subordinates did not. Ruff colour of male Himalayan tahr may have evolved as a rank signal, homologous to horn size in wild sheep and goats.

Intolerance amongst deer species at feeding: roe deer are uneasy banqueters.

The function of inter-specific interactions in large herbivore assemblies is under debate. We have studied inter-specific interactions between roe and fallow deer on pastures, to assess whether competition between these Cervidae occurs through behavioural interference. Roe were displaced by fallow deer (i.e. the former moved away from the latter at a distance of >50 m) in 83% of cases (N=127). Ninety-four percentage (N=83) of displacement events occurred while roe deer were feeding; in 50% of these cases (N=78), roe stopped grazing and left the feeding ground. Even when fallow deer did not show any sign of direct aggression to roe, these moved away from fallow in 72% of cases (N=127). Vigilance rate was significantly greater in roe than in fallow deer, irrespectively of the presence of the other species. When roe and fallow deer grazed within 50 m from each other, vigilance rate increased significantly in roe, but not in fallow deer. Roe deer, in a group, were significantly more tolerant of the presence of fallow deer (even in group), than when solitary. Fallow deer seemed to be able to exclude roe deer from feeding sites through behavioural intolerance.

Immobilization of Himalayan tahr with a xylazine-ketamine mixture and reversal with atipamezole under field conditions.

Twenty-nine free-ranging Himalayan tahr (Hemitragus jemlahicus) were darted in the Sagarmatha National Park (Nepal) using different combinations of xylazine and ketamine. Animals in Group 1 (n = 4) received a mean xylazine-ketamine dose of 2.77 +/- 0.99 mg/kg xylazine plus 3.32 +/- 0.19 mg/kg ketamine in males and 2.39 +/- 0.10 mg/kg xylazine plus 4.29 +/- 0.17 mg/kg ketamine in females. Animals in Group 2 (n = 25) received a mean xylazine-ketamine dose of 1.70 +/- 0.41 mg/kg xylazine plus 3.06 +/- 0.74 mg/kg ketamine in males and 1.82 +/- 0.29 mg/kg xylazine plus 3.29 +/- 0.52 mg/kg ketamine in females. No anesthetic-related mortality was recorded. Anesthesia was reversed by a standard dose of 11 mg/animal of atipamezole administered by intramuscular injection. Although all anesthetic dosages immobilized free-ranging tahr successfully, a quick and smooth recovery was obtained (11.1 +/- 5.6 min) only with the dosages of Group 2.

The interacting RNA polymerase II subunits, hRPB11 and hRPB3, are coordinately expressed in adult human tissues and down-regulated by doxorubicin.

We previously isolated the human RPB11 cDNA, encoding the 13.3 kDa subunit of RNA polymerase II, and demonstrated that expression of this subunit is modulated by doxorubicin. Using hRPB11 as bait in a yeast two-hybrid system, two cDNA variants encoding a second RNA polymerase II subunit, hRPB3, have now been isolated and characterized. These two hRPB3 mRNA species differed in 3' UTR region length, the longer transcript containing the AU-rich sequence motif that mediates mRNA degradation. Both hRPB11 and hRPB3 transcripts share a similar pattern of distribution in human adult tissues, with particularly high levels in both heart and skeletal muscle, and the expression of both is down-regulated by doxorubicin as found previously for the hRPB11 subunit. Taken together, these findings suggest that the interaction between hRPB3 and hRPB11 is fundamental for their function and that this heterodimer is involved in doxorubicin toxicity.

A simple method for non-radioactive PCR-SSCP using MDE gel solution and a midi gel format: application for the detection of variants in the GLUT1 and CTLA-4 genes.

The need to identify disease-causing mutations and DNA polymorphisms has increased with the continuing identification of new candidate genes. PCR single-strand conformation polymorphism (PCR-SSCP) is one of the techniques most widely used to identify a mutant sequence or a polymorphism in a known gene. However, the original SSCP protocols using the incorporation of radioactive label and polyacrylamide gel electrophoresis on sequencing gels for detection were labour intensive and time-consuming. Here we describe a simple SSCP protocol using MDE gel solution and a midi gel format to detect SSCP variations in the glucose transporter gene GLUT1, that we have previously analysed with the standard radioactive SSCP protocol, and we have also tested this method on the previously described point mutation (A/G transition in exon 1) of the CTLA-4 (cytotoxic T lymphocyte associated-4) gene. All known variants were detected. Based on the results, this technique appears to be simple, with no use of radioactive labels and with easy handling of the gel. Furthermore, it needs little optimisation, is relatively rapid and highly sensitive. We propose this method for the first screening for candidate gene variants.

Habitat richness affects home range size in the red fox Vulpes vulpes.

The spatial behaviour of the red fox Vulpes vulpes shows a great flexibility (Voigt and Macdonald, 1984). Home range size varies from 10 to over 5000 ha (Macdonald, 1987; Voigt, 1987). In carnivores, variations in home range size, weighed for body mass (Gittleman and Harvey, 1982), are largely related to differences in habitat productivity, but the intraspecific local variation in home range size can be only partially explained by differences in productivity. Macdonald (1981, 1983) suggested that home range size and configuration were determined in the red fox by the dispersion of food-rich patches. More recently, it has been found that In this paper, evidence for a strong influence of habitat richness on home range size of the red fox is reported.

Excursion behaviour of female roe deer may depend on density.

The excursion behaviour of roe does was studied for two years in a low density population (ca. 6.5 ind./100 ha), in an Apennine-continental forest of Southern Italy, through satellite radiotracking. During the rutting period, our radiotagged does (N=6) moved well outside their summer ranges, with an average exploration area eight times greater than summer ranges. The median duration of excursions was 51 h (range: 10-99 h). One female stayed away for a total of 11 days. In order further to understand this behaviour, we reviewed all studies (N=6) dealing with excursion behaviour of roe does and carried out in areas where population density was assessed through the same method (i.e. drive counts). Out of five ecological parameters included in the analysis, excursion behaviour of roe does was found significantly and negatively associated only to population density: when density was low, the proportion of roaming does increased, probably because of the lower availability of "free" bucks during the short time of female oestrous.

Mating dominance amongst male Himalayan tahr: blonds do better.

In ungulates, rank order is determined by differences in weight, body size, weapon size and age. In the Caprini tribe (Bovidae: Caprinae), adult male Himalayan tahr are unique to show different coat colours, but no sexual dimorphism in weapons. A highly significant correlation between hair colour and rank order was found during the rut: males with a lighter coloured ruff dominated over darker ruffed ones, in both aggressive interactions and access to oestrus females. We studied colour-based dominance in relation to weight, age and testosterone levels, which establish the social rank in most ungulates. No differences in weight and testosterone concentrations were found between adult male colour classes, but males with paler ruffs were significantly younger than darker adult males. The distribution of physical traumas from fights confirmed that younger, lighter-coloured males had a higher rank than older, darker males, a pattern which is unusual amongst ungulates. Coat colour seems to work as a signal of rank in male-male aggressive interactions and it changes according to age, whereas the relevant physiological determinants deserve further research. Intrasexual male competition has not changed weapon size or shape in the Himalayan tahr, but ruff colours are apparently used to signal rank and dominance. Colour patterns of adult males may then be homologous to ritualised weapons, apparently being a unique feature of male tahr amongst mammals.

Prevalence and concentration of Verotoxigenic Escherichia coli O157:H7 in adult sheep at slaughter from Italy.

A 1-year study on the animal-level prevalence and concentration of Escherichia coli O157 in adult sheep at slaughter was performed, to collect qualitative and quantitative information on the diffusion of the pathogen in adult sheep from Italy. A total 533 samples were collected, with a similar distribution in the four seasons. For prevalence estimates, a simple random sampling technique was used. An immuno-magnetic separation technique was used for sample screening, with enumeration of the pathogen in positive samples, along with molecular and serological identification of isolates. An overall prevalence of 7.1% (38/ 533, 95% CI 4.9-9.3%) was observed for fully virulent E. coli O157. A wide interval of VTEC O157 per gram was observed (< 100 to 6 x 10(5) CFU g(-1)), with 28.9% (11/38) of positive samples > or = 1 x 10(3) CFU g(-1), set as the threshold for those animals defined 'active shedders' for the purpose of the study. Eight per cent (3/38) of animals shed > 1 x 10(4) g(-1) VTEC O157, which represents > 96% of the total VTEC O157 bacteria cultured from all animals tested. The prevalence estimate of active shedders was therefore 2.1% (95% CI 0.9-3.3%). Most (34/38, 89.5%) of the positive animals were found in summer (July-September). Prevalence and concentrations of virulent VTEC O157 obtained in this study contribute to the demonstration that adult sheep represent a relevant source of environmental contamination from virulent VTEC O157, as well as a source of VTEC O157 contamination for food of ovine origin (meat and dairy products), especially during warm months.

Prevalence of Escherichia coli O157 in lambs at slaughter in Rome, central Italy.

A study on the prevalence of the faecal carriage of Escherichia coli O157 in lambs was performed in the major slaughterhouse in Rome, central Italy, during 2002. A total of 643 animals, consisting of 378 weaned and 265 suckling lambs, were assayed for the presence of E. coli O157. Five O157-agglutinating E. coli strains were isolated (0.8%, 95% CI 0.3-1.9). Only one was positive to PCR specific for the eae gene and produced verocytotoxin VT2, with a VTEC O157 overall prevalence of 0.2% (95% CI 0.0-1.0), whereas one strain possessed the eae gene only. All the other isolates were negative for the presence of all the virulence genes considered. The animals were either from local farms or imported from Eastern Europe. The results suggest an age-specific difference since the microorganism was isolated only from 0.3% (95% CI 0.0-1.7) of weaned lambs, while all samples from suckling lambs tested negative. From this study, the overall risk of human exposure to pathogenic E. coli O157 from lamb meat consumption derived from the major slaughterhouse in Rome can be considered reasonably low, particularly when suckling lamb meat is considered.

How can we use the red fox as a bioindicator of organochlorines?

The exposure of red foxes to hexachlorobenzene (HCB), dichlorodiphenyltrichloroethan (DDT), DDT derivatives, and polychlorinated biphenyls (PCBs) in central Italy was determined by analyzing muscle and adipose tissue samples from 57 red foxes collected in 1992-1993 in three areas of Tuscany, each characterized by different levels of anthropization. Chemical analysis showed that HCB occurred in low concentrations, ranging from an average of 0.08 microg/g lipid basis in fat of foxes from Pisa, the least agricultural area, to 1.06 microg/g in muscle of those from Grosseto, the least industrialized area. DDTs were calculated as the sum of the op' and pp'DDT isomer concentrations and the DDT derivatives DDE and DDD. The lowest values of DDTs were found in foxes from Pisa, with mean values in muscle of 0.55 microg/g and 0.14 microg/g in fat. In the outskirts of Siena, the corresponding values were 1.16 microg/g and 0.49 microg/g. Higher values were found in the Grosseto area: 4.17 microg/g in muscle and 0.67 microg/g in fat. The most abundant DDT was pp'DDE in all areas, constituting more than half of total DDTs. Mean and standard deviations of PCB concentrations (calculated as the sum of the 31 identified and quantified congeners), were 20.2 microg/g in muscle and 7.2 microg/g in fat of red foxes from the Siena area. Values in the Pisa area were lower (7. 6 microg/g in muscle and 1.8 microg/g in fat), whereas the highest levels were found in the Grosseto area (38 microg/g in muscle and 7. 5 microg/g in fat). PCB fingerprints showed that hexa- and heptachlorobiphenyls were the most common congeners, although amounts differed between the three localities and the two tissues considered. As far as PCB composition is concerned, detailed observation revealed that although the three areas highly differed between each other, the most abundant congeners were the 22'344'5' (PCB 138) and 22'44'55' (PCB 153) hexachlorbiphenyls and 22'33'44'5 (PCB 170) and 22'344'55' (PCB 180) heptachlorobiphenyls; these accounted for over 60% of total PCB residues in muscle and fat. Concentrations of the remaining PCB congeners were all lower than 5% of total PCBs, with the exception of octachlorobiphenyl 22'33'44'55' (PCB 194) in fat tissue. Foxes may be suitable biological indicators to detect organochlorine contamination in the environment because of their catholic food habits. This information may be used to select specific bioindicators for assessing the relative level of pollutants in that area.

The G972R variant of the insulin receptor substrate-1 (IRS-1) gene, body fat distribution and insulin-resistance.

AIMS/HYPOTHESIS: Insulin resistance is recognised as the core factor in the pathogenesis of Type II (non-insulin-dependent) diabetes mellitus, hypertension and atherosclerosis. Several studies indicate the possible role of mutations of the insulin receptor substrate-1 (IRS-1) gene in the pathogenesis of insulin-resistance and suggest a possible interaction between the IRS-1 gene and obesity, either by an effect on the development of obesity or by causing or aggravating the obesity-associated insulin resistance. Therefore, the prevalence of the G972R mutation of the IRS-1 gene was compared in 157 non-diabetic obese subjects (BMI > 30 m/kg2) and in 157 lean subjects (BMI < 28 m/kg2). By investigating the relation between this IRS-1 mutation, measures of obesity and metabolic parameters, we explored the possible influence of this mutation on body fat distribution and insulin resistance. METHODS: The G972R mutation was detected by PCR amplification and BstN-1 restriction enzyme digestion. Data were analysed by univariate and multivariate analysis. RESULTS: The G972R allele was significantly more frequent in obese subjects than in lean subjects (p < 0.002); however, no difference was found between centrally and peripherally obese subjects. Obese G972R carriers had significantly higher BMI (p < 0.001), fasting insulin (p < 0.001), triglycerides (p < 0.03) and HOMAIR (p < 0.001) than obese non-carriers. No differences were observed between G972R carriers and non-carriers among control subjects. Multivariate analysis confirmed that the IRS-1 G972R mutation was significantly and independently associated with reduced insulin sensitivity (p < 0.009) in the obese group. CONCLUSION/INTERPRETATION: The G972R mutation of the IRS-1 gene associates with obesity, but not with fat distribution, in this Italian cohort, and within the obese subjects this IRS-1 variant strongly associates with metabolic parameters suggesting greater insulin-resistance. These findings indicate a possible interaction between the IRS-1 variant and obesity in worsening insulin sensitivity.

A common mutation of the insulin receptor substrate-1 gene is a risk factor for coronary artery disease.

Insulin resistance is associated with increased risk of atherosclerosis. Insulin receptor substrate-1 (IRS-1) plays a key role in tissue insulin sensitivity. A common mutation (G972R) of the IRS-1 gene has been shown to impair IRS-1 function, and it has been associated with reduced insulin sensitivity and lipid abnormalities. This led us to investigate the role of the G972R mutation in predisposing individuals to coronary artery disease (CAD). The DNA of 318 subjects with angiographically documented coronary atherosclerosis (>50% stenosis) and 208 population control subjects was analyzed for the presence of the G972R mutation. This mutation was detected by nested polymerase chain reaction and BstNI restriction enzyme digestion. The frequency of the G972R mutation was significantly higher among patients with CAD than controls (18. 9% versus 6.8%, respectively; P<0.001). After controlling for other coronary risk factors, the relative risk of CAD associated with the G972R mutation was 2.93 (95% CI 1.30 to 6.60; P<0.02) in the entire cohort. This risk was found to be even higher in the subgroups of obese subjects (odds ratio [OR] 6.97, 95% CI 2.24 to 21.4; P<0.001) and subjects with clinical features of insulin resistance syndrome (OR 27.3, 95% CI 7.19 to 104.0; P<0.001). The IRS-1 gene variant was associated with a higher frequency of diabetes mellitus (14.9% among carriers versus 6.5% among noncarriers; P<0.01) and with a 60% increase of plasma total triglycerides (P<0.001). Also, plasma concentrations of total cholesterol and the ratio of total cholesterol to HDL cholesterol were significantly (P<0.001) higher among carriers than noncarriers, although to lesser a extent. These effects were independent of CAD status. The G972R mutation in the IRS-1 gene was found to be a significant independent predictor of CAD. Moreover, this mutation greatly increased the risk of CAD in obese subjects and in patients with the cluster of abnormalities of insulin resistance syndrome. Besides the increased frequency of diabetes, carriers showed a more atherogenic lipid profile, suggesting a potential role of the IRS-1 gene in the pathogenesis of lipid abnormalities associated with CAD.