Genetic resistance determinants to fusidic acid and chlorhexidine in variably susceptible staphylococci from dogs.

BACKGROUND: Concern exists that frequent use of topically-applied fusidic acid (FA) and chlorhexidine (CHX) for canine pyoderma is driving clinically relevant resistance, despite rare description of FA and CHX genetic resistance determinants in canine-derived staphylococci. This study aimed to determine minimum inhibitory concentrations (MICs) and investigate presence of putative resistance determinants for FA and CHX in canine-derived methicillin-resistant (MR) and -susceptible (MS) staphylococci. Plasmid-mediated resistance genes (fusB, fusC, fusD, qacA/B, smr; PCR) and MICs (agar dilution) of FA and CHX were investigated in 578 staphylococci (50 MR S. aureus [SA], 50 MSSA, 259 MR S. pseudintermedius [SP], 219 MSSP) from Finland, U.S.A., North (NUK) and South-East U.K. (SEUK) and Germany. In all isolates with FA MIC ≥64 mg/L (n = 27) fusA and fusE were amplified and sequenced. RESULTS: FA resistance determinants (fusA mutations n = 24, fusB n = 2, fusC n = 36) were found in isolates from all countries bar U.S.A. and correlated with higher MICs (≥1 mg/L), although 4 SP isolates had MICs of 0.06 mg/L despite carrying fusC. CHX MICs did not correlate with qacA/B (n = 2) and smr (n = 5), which were found in SEUK SA, and SP from NUK and U.S.A. CONCLUSIONS: Increased FA MICs were frequently associated with fusA mutations and fusC, and this is the first account of fusB in SP. Despite novel description of qacA/B in SP, gene presence did not correlate with CHX MIC. Selection pressure from clinical use might increase prevalence of these genetic determinants, but clinical significance remains uncertain in relation to high skin concentrations achieved by topical therapy.

Non-tuberculous Mycobacteria can Cause Disseminated Mycobacteriosis in Cats.

Mycobacteriosis caused by non-tuberculous mycobacteria (NTM) is a rising concern in human medicine both in immunocompromised and immunocompetent patients. In cats, mycobacteriosis caused by NTM is considered mostly to be a focal or dermal infection, with disseminated disease mostly caused by Mycobacterium avium. We describe three cases of disseminated mycobacteriosis in cats, caused by Mycobacterium malmoense, Mycobacterium branderi/shimoidei and M. avium, with no identified underlying immunosuppression. In all cases, extracellular mycobacteria were seen in the pulmonary epithelium, intestinal lumen and glomerular tufts, which could affect the shedding of the organism. The present study highlights the importance of mycobacteriosis as a differential even in immunocompetent animals. Considering the close relationship of owners and pets and the potential presence of free mycobacteria in secretions, cats should be considered as a possible environmental reservoir for mycobacteria.

Sex-specific compensatory growth in the larvae of the greater wax moth Galleria mellonella.

Deficiency of food resources in ontogeny is known to prolong an organism's developmental time and affect body size in adulthood. Yet life-history traits are plastic: an organism can increase its growth rate to compensate for a period of slow growth, a phenomenon known as 'compensatory growth'. We tested whether larvae of the greater wax moth Galleria mellonella can accelerate their growth after a fast of 12, 24 or 72 h. We found that a subgroup of female larvae showed compensatory growth when starved for 12 h. Food deficiency lasting more than 12 h resulted in longer development and lower mass gain. Strength of encapsulation reactions against a foreign body inserted in haemocoel was the weakest in females that showed compensatory growth, whereas the strongest encapsulation was recorded in the males and females that fasted for 24 and 72 h. More specifically, we found sex-biased immune reactions so that females had stronger encapsulation rates than males in one group that fasted for 72 h. Overall, rapidly growing females had a short larval development period and the shortest adult lifespan. These results suggest that highly dynamic trade-offs between the environment, life-history traits and sex lead to plasticity in developmental strategies/growth rates in the greater wax moth.

The masculinity paradox: facial masculinity and beardedness interact to determine women's ratings of men's facial attractiveness.

In many species, male secondary sexual traits have evolved via female choice as they confer indirect (i.e. genetic) benefits or direct benefits such as enhanced fertility or survival. In humans, the role of men's characteristically masculine androgen-dependent facial traits in determining men's attractiveness has presented an enduring paradox in studies of human mate preferences. Male-typical facial features such as a pronounced brow ridge and a more robust jawline may signal underlying health, whereas beards may signal men's age and masculine social dominance. However, masculine faces are judged as more attractive for short-term relationships over less masculine faces, whereas beards are judged as more attractive than clean-shaven faces for long-term relationships. Why such divergent effects occur between preferences for two sexually dimorphic traits remains unresolved. In this study, we used computer graphic manipulation to morph male faces varying in facial hair from clean-shaven, light stubble, heavy stubble and full beards to appear more (+25% and +50%) or less (-25% and -50%) masculine. Women (N = 8520) were assigned to treatments wherein they rated these stimuli for physical attractiveness in general, for a short-term liaison or a long-term relationship. Results showed a significant interaction between beardedness and masculinity on attractiveness ratings. Masculinized and, to an even greater extent, feminized faces were less attractive than unmanipulated faces when all were clean-shaven, and stubble and beards dampened the polarizing effects of extreme masculinity and femininity. Relationship context also had effects on ratings, with facial hair enhancing long-term, and not short-term, attractiveness. Effects of facial masculinization appear to have been due to small differences in the relative attractiveness of each masculinity level under the three treatment conditions and not to any change in the order of their attractiveness. Our findings suggest that beardedness may be attractive when judging long-term relationships as a signal of intrasexual formidability and the potential to provide direct benefits to females. More generally, our results hint at a divergence of signalling function, which may result in a subtle trade-off in women's preferences, for two highly sexually dimorphic androgen-dependent facial traits.

Intra- and trans-generational effects of larval diet on susceptibility to an entomopathogenic fungus, Beauveria bassiana, in the greater wax moth, Galleria mellonella.

In addition to nutritional conditions experienced by individuals themselves, those experienced by their parents can affect their immune function. Here, we studied the intra- and trans-generational effects of larval diet on susceptibility to an entomopathogenic fungus, Beauveria bassiana, in the greater wax moth, Galleria mellonella. In the first part of the study, a split-brood design was used to compare the susceptibility of full sibs raised either on low- or on high-nutrition larval diet. In the second part of the study, a similar experimental design was employed to investigate the effects of maternal and paternal diet as well as their interaction on offspring's susceptibility. In the first part of the study, we found that individuals fed with high-nutrition diet had higher mortality from infection than individuals fed with low-nutrition diet. However, diet did not affect post-infection survival time. Conversely, in the second part of the study, maternal diet was found to have no significant effect on final mortality rate of offspring, but it affected survival time: larvae with high-nutrition maternal diet survived fewer days after infection than larvae with low-nutrition maternal diet. Paternal diet had no significant effect on offspring's susceptibility to the fungus, indicating that paternal effects are not as important as maternal effects in influencing immune function in this species. Our findings provide further indication that maternal nutrition affects immune function in insects, and suggest that the direct effects of nutrition on immunity may be different, yet parallel, to those caused by parental nutrition.

Male mealworm beetles increase resting metabolic rate under terminal investment.

Harmful parasite infestation can cause energetically costly behavioural and immunological responses, with the potential to reduce host fitness and survival. It has been hypothesized that the energetic costs of infection cause resting metabolic rate (RMR) to increase. Furthermore, under terminal investment theory, individuals exposed to pathogens should allocate resources to current reproduction when life expectancy is reduced, instead of concentrating resources on an immune defence. In this study, we activated the immune system of Tenebrio molitor males via insertion of nylon monofilament, conducted female preference tests to estimate attractiveness of male odours and assessed RMR and mortality. We found that attractiveness of males coincided with significant down-regulation of their encapsulation response against a parasite-like intruder. Activation of the immune system increased RMR only in males with heightened odour attractiveness and that later suffered higher mortality rates. The results suggest a link between high RMR and mortality and support terminal investment theory in T. molitor.

Serum C-reactive protein as a diagnostic biomarker in dogs with bacterial respiratory diseases.

BACKGROUND: C-reactive protein (CRP) is a major acute-phase protein in dogs. Serum concentrations are low in healthy animals, but increase rapidly after inflammatory stimuli. OBJECTIVE: The aim of the study was to investigate CRP concentrations in various respiratory diseases of dogs and to determine if CRP can be used as a biomarker in the diagnosis of bacterial respiratory diseases. ANIMALS: A total of 106 privately owned dogs with respiratory diseases (17 with bacterial tracheobronchitis [BTB], 20 with chronic bronchitis [CB], 20 with eosinophilic bronchopneumopathy [EBP], 12 with canine idiopathic pulmonary fibrosis [CIPF], 15 with cardiogenic pulmonary edema [CPE], and 22 with bacterial pneumonia [BP]) and 72 healthy controls. METHODS: The study was conducted as a prospective cross-sectional observational study. CRP was measured in serum samples. Diagnosis was confirmed by clinical and laboratory findings, diagnostic imaging, and selected diagnostic methods such as cytological and microbiological analysis of respiratory samples, echocardiography, and histopathology. RESULTS: Dogs with BP had significantly higher CRP concentrations (median, 121 mg/L; interquartile range, 68-178 mg/L) than dogs with BTB (23, 15-38, P = .0003), CB (13, 8-14, P < .0001), EBP (5, 5-15, P < .0001), CIPF (17, 10-20, P < .0001), or CPE (19, 13-32, P < .0001) and healthy controls (14, 8-20, P < .0001). Dogs with BTB had significantly higher CRP concentrations than dogs with CB (P = .001) or EBP (P < .0001) and healthy controls (P = .029). CONCLUSION AND CLINICAL IMPORTANCE: These results indicate that CRP has potential for use as an additional biomarker, especially in the diagnostics of BP.

Genetic and phenotypic relationships between immune defense, melanism and life-history traits at different temperatures and sexes in Tenebrio molitor.

Insect cuticle melanism is linked to a number of life-history traits, and a positive relationship is hypothesized between melanism and the strength of immune defense. In this study, the phenotypic and genetic relationships between cuticular melanization, innate immune defense, individual development time and body size were studied in the mealworm beetle (Tenebrio molitor) using three different temperatures with a half-sib breeding design. Both innate immune defense and cuticle darkness were higher in females than males, and a positive correlation between the traits was found at the lowest temperature. The effect of temperature on all the measured traits was strong, with encapsulation ability and development time decreasing and cuticle darkness increasing with a rise in temperature, and body size showing a curved response. The analysis showed a highly integrated system sensitive to environmental change involving physiological, morphological and life-history traits.

Cross-cultural variation in women's preferences for cues to sex- and stress-hormones in the male face.

Women in the UK prefer the faces of men with low levels of the stress hormone cortisol, and the relationship is moderated by the sex hormone testosterone. In a Latvian sample, however, women's preferences were not affected by cortisol, and the interaction with testosterone differed from that of the UK. To further explore cross-cultural variation in preferences for facial cues to sex- and stress-hormones, we tested the preferences of women from 13 countries for facial composites constructed to differ in combinations of the hormones. We found significant relationships between a measure of societal development (the United Nations human development index 2011) and preferences for cues to testosterone in the face, and the interaction between preferences for cues to testosterone and cortisol. We also found a significant relationship between preferences for cues to testosterone and a societal-level measure of parasite stress. We conclude that societal-level ecological factors influence the relative value of traits revealed by combinations of sex- and stress-hormones.

Senescence in immune priming and attractiveness in a beetle.

Age-related decline in immune activity is referred to as immunosenescence and has been observed for both the adaptive immune response of vertebrates and the innate immune system of invertebrates. Because maintaining a basic level of immune defence and mounting an immune response is costly, optimal investment in immune function should vary over a wide range of individual states such as the individual's age. In this study, we tested whether the immune response and immunological priming within individuals become less efficient with age using mealworm beetles, Tenebrio molitor, as a model organism. We also tested whether ageing and immunological priming affected the odours produced by males. We found that young males of T. molitor were capable of mounting an immune response a sterile nylon monofilament implant with the potential to exhibit a simple form of immune memory through mechanisms of immune priming. Older males did not increase their immune response to a second immune challenge, which negatively affected their sexual attractiveness and remaining life span. Our results indicate that the immune system of older males in T. molitor is less effective, suggesting complex evolutionary trade-offs between ageing, immune response and sexual attractiveness.

There is more than one way to skin a G matrix.

Because of its importance in directing evolutionary trajectories, there has been considerable interest in comparing variation among genetic variance-covariance (G) matrices. Numerous statistical approaches have been suggested but no general analysis of the relationship among these methods has previously been published. In this study, we used data from a half-sib experiment and simulations to explore the results of applying eight tests (T method, modified Mantel test, Bartlett's test, Flury hierarchy, jackknife-manova, jackknife-eigenvalue test, random skewers, selection skewers). Whereas a randomization approach produced acceptable estimates, those from a bootstrap were typically unacceptable and we recommend randomization as the preferred method. All methods except the jackknife-eigenvalue test gave similar results although a fine-scale analysis suggested that the former group can be subdivided into two or possibly three groups, hierarchical tests, skewers and the rest (jackknife-manova, modified Mantel, T method, probably Bartlett's). An advantage of the jackknife methods is that they permit tests of association with other factors, such as in this case, temperature and sex. We recommend applying all the tests described in this article, with the exception of the T method, and provide R functions for this purpose.

Female choice reveals terminal investment in male mealworm beetles, Tenebrio molitor, after a repeated activation of the immune system.

Increasing evidence suggests that secondary sexual traits reflect immunocompetence of males in many animal species. This study experimentally investigated whether a parasite-like immunological challenge via a nylon implant affects sexual attractiveness of males in Tenebrio molitor L. (Coleoptera: Tenebrionidae) Although a single immunological challenge significantly reduced sexual attractiveness and locomotor activity of males, it had no adverse effect on their survival. A second immune challenge of the same males increased their attractiveness. However, it was found that the repeated challenge significantly reduced locomotor activity of males and caused higher mortality. This result indicates terminal investment on sexual signaling, which is supposedly based on a trade-off between pheromone production and energy expenditures needed for such activities as recovery of immune system and locomotor activity. When the third implantation was carried out in the same group of males, melanization of nylon implants was found to be lower in more attractive than in less attractive males. This suggests that males that became sexually attractive after the second immune challenge did not invest in recovery of their immune system.

The effects of dietary nickel on the detoxification enzymes, innate immunity and resistance to the fungus Beauveria bassiana in the larvae of the greater wax moth Galleria mellonella.

In this study, we tested the effects of dietary nickel on the activity of glutathione S-transferase (GST), esterases, phenoloxidase, and encapsulation in the haemolymph of larvae of the greater wax moth Galleria mellonella. We also explored the effects of dietary nickel on larval resistance to infection by the fungus Beauveria bassiana. Larvae fed a low dose of nickel (10 µg g(-1)) had significantly higher GST, phenoloxidase activity and encapsulation responses than controls fed on a nickel-free diet. We also found that larvae fed a sublethal dose of nickel (50 µg g(-1)) had increased GST, esterase activity and encapsulation rates but decreased phenoloxidase activity. Although, a sublethal dose of dietary nickel enhanced innate immunity, we found that this reduced resistance against the real pathogen. Our results suggest that enhanced immunity and detoxification enzyme activity of insects may not be beneficial to resistance to fungal infection. It appears that there is a trade off between different resistance mechanisms in insects under different metal treatments.

Analysis of the effects of early nutritional environment on inbreeding depression in Drosophila melanogaster.

The impact of nutritional deficiencies early in life in determining life-history variation in organisms is well recognized. The negative effects of inbreeding on fitness are also well known. Contrary to studies on vertebrates, studies on invertebrates are not consistent with the observation that inbreeding compromises resistance to parasites and pathogens. In this study, we investigated the effect of early nutrition on the magnitude of inbreeding depression in development time, adult body size and adult resistance to the bacterium Serratia marcescens in Drosophila melanogaster. We found that early nutritional environment had no effect on the magnitude of inbreeding depression in development time or adult body size but may have played a small role in adult resistance to the bacterial infection. Estimates of heritabilities for development time under the poor nutritional environment were larger than those measured under the standard nutritional conditions.

The effect of a short-term hypocaloric diet on liver gene expression and metabolic risk factors in obese women.

BACKGROUND AND AIMS: Most gene expression studies examining the effect of obesity and weight loss have been performed using adipose tissue. However, the liver also plays a central role in maintaining energy balance. We wanted to study the effects of a hypocaloric diet on overall hepatic gene expression and metabolic risk factors. METHODS AND RESULTS: The study subjects were middle-aged, obese women. The diet intervention subjects (n=12) were on a hypocaloric, low-fat diet for 8 weeks with a daily energy intake of 5.0 MJ (1200 kcal), while the control subjects (n=19) maintained their weight. Liver biopsies were taken at the end of the diet period during a gallbladder operation. Hepatic gene expression was analyzed using microarrays by comparing the gene expression profiles from four subjects per group. A global decrease in gene expression was observed with 142 down-regulated genes and only one up-regulated gene in the diet intervention group. The diet resulted in a mean weight loss of 5% of body weight. Triglyceride and fasting insulin concentrations decreased significantly after the diet. CONCLUSIONS: The global decrease in hepatic gene expression was unexpected but the results are interesting, since they included several genes not previously linked to weight reduction. However, since the comparison was made only after the weight reduction, other factors in addition to weight loss may also have been involved in the differences in gene expression between the groups. The decrease in triglyceride and fasting plasma insulin concentrations is in accordance with results from previous weight-loss studies.

Effect of GnRH dose on occurrence of short oestrous cycles and LH response in cyclic dairy heifers.

Prostaglandin F(2 alpha) (PGF(2 alpha)) and GnRH treatments given 24 h apart have been shown to result in short oestrous cycles (8-12 days) in some cows and heifers. The differences in responses may depend on the dose of GnRH. Therefore, the effect of the dose of GnRH on occurrence of short cycles and LH response was studied here. Oestrus was induced with dexcloprostenol (0.15 mg) in two groups of Ayrshire heifers. A second luteolysis was induced similarly on day 7 after ovulation; 24 h after PGF(2 alpha) treatment, the heifers were administered either a high (0.5 mg, n = 15, group T500) or low (0.1 mg, n = 10, group T100) dose of gonadorelin. Blood samples for progesterone analyses were collected daily from the second PGF(2 alpha) administration to the second ovulation after the PGF(2 alpha) injection. Beginning 24 h after the GnRH treatment, ovaries were examined by transrectal ultrasonography every 6 h until ovulation, and daily between day 4 and the next ovulation. Five heifers from both groups were sampled for LH analyses via a jugular catheter every 30 min from 1 h before to 6 h after the GnRH administration. Short oestrous cycles were detected in 7 of 10 cases in group T100 and in 12 of 15 cases in group T500. No significant differences in LH responses were detected between the groups. In group T500, the rise in LH concentration tended to be somewhat slower than in group T100. The dose of GnRH (0.1 vs 0.5 mg) did not affect the occurrence of short oestrous cycles and LH response.

Severe hypoglycemia during intensive insulin therapy.

BACKGROUND: Tight glycemic control reduces mortality in surgical intensive care patients and in long-term medical intensive care patients. A large study on intensive insulin therapy was prematurely discontinued due to safety issues. As the safety of intensive insulin therapy has been questioned, we screened all patients during a 17-month period to reveal the incidence of hypoglycemia and its effects on the outcome of the patients. METHODS: All patients treated between February 2005 and June 2006 in two intensive care units (ICUs) of a tertiary care teaching hospital were included in the study. A nurse-driven intensive insulin therapy with a target blood glucose level of 4-6 mmol/l had been introduced earlier. The patients were divided into two groups according to the presence of severe hypoglycemia (

Effect of time interval between prostaglandin F(2alpha) and GnRH treatments on occurrence of short estrous cycles in cyclic dairy heifers and cows.

Prostaglandin F(2alpha) (PGF(2alpha)) and GnRH treatments, when administered 24h apart during early diestrus, cause short estrous cycles in some dairy cows and heifers [J. Taponen, M. Kulcsar, T. Katila, L. Katai, G. Huszenicza, H. Rodriguez-Martinez, Short estrous cycles and estrous signs after premature ovulations induced with cloprostenol and gonadotropin-releasing hormone in cyclic dairy cows, Theriogenology 2002; 58, 1291-1302]. We investigated the effect of a time interval between PGF(2alpha) and GnRH administration on the appearance of short cycles. Estrus was induced in heifers with dexcloprostenol. A second luteolysis was induced similarly on day 7 after ovulation, and either 0 (T0) or 24h (T24) later an injection of GnRH (0.1mg of gonadorelin) was administered. We monitored ovarian activity with progesterone analyses from blood plasma samples and with ultrasonography. Fourteen cases (12 in T0 and 2 in T24) were excluded due to either incomplete luteolysis (2 cases) or unresponsiveness to GnRH (10 in T0 and 2 in T24). Short estrous cycles (7 to 8 d) were detected in 11/11 and 8/17 heifers in groups T0 and T24, respectively, with a significant difference in the incidence of short cycles (P<0.01). In Experiment 2, estrus was induced in cows on day 8 (D8, n=18), 9 (D9, n=5), or 10 (D10, n=3) with cloprostenol and gonadorelin administered simultaneously. Daily milk samples were collected for progesterone analysis until subsequent estrus was detected and ovarian ultrasound examinations were performed. Eight cases had to be excluded due to unresponsiveness to GnRH, leaving 18 cases eligible for the study. Short estrous cycles (7-12d) were detected in 14/18 cows. In conclusion, shortening the time interval between PGF(2alpha) and GnRH treatments increased the incidence of short estrous cycles and appeared to increase the proportion of females unresponsive to GnRH treatment.

Effect of bacterial infection on antioxidant activity and lipid peroxidation in the midgut of Galleria mellonella L. larvae (Lepidoptera, Pyralidae).

Bacillus thuringiensis is one of the most widely used sources of biorational pesticides, as well as a key source of genes for transgenic expression to provide pest resistance in plants. In this study the effect of Bacillus thuringiensis ssp. galleriae (Bt) infection on the activity of superoxide dismutase (SOD), glutathione S-transferase (GST), catalase (CAT), concentrations of oxidated and reduced thiols (RSSR/RSH) and malondialdehyde (MDA) was tested in the midgut of Galleria mellonella larvae. We found that Bt infection resulted in increased activities of SOD, GST, malondialdehyde and RSSR/RSH ratio the first day after inoculation. However, catalase activity decreased on the first and following days after bacterial infection by Bt. Our results confirm the hypothesis that Bt infection increases the level of oxidative stress in the larval midgut. In light of this study, it seems possible that oxidative damage contributes to cell death in the midgut during bacteriosis.