Incidence of Bacterial Chondronecrosis with Osteomyelitis (Femoral Head Necrosis) Induced by a Model of Skeletal Stress and its Correlation with Subclinical Necrotic Enteritis.

Bacterial chondronecrosis with osteomyelitis (BCO) is a septic necrosis of the skeletal system of unknown origin and an important cause of lameness in broiler chickens. Epithelial inflammation has been proposed as an avenue for bacterial translocation leading to BCO. We evaluated the effect of subclinical necrotic enteritis (SNE), an intestinal inflammatory event, with the development of BCO. In each of two experiments, chickens were divided into three treatments: 1) SNE challenge, including both dietary (wheat- and fish-based diet) and intestinal pathogenic challenges (Eimeria maxima and Clostridium perfringens), 2) dietary challenge only, and 3) control diet (corn- and soy-based diet). Floor ramps were introduced as part of an established method for increasing the frequency of BCO. The efficacy of the SNE challenge was corroborated by necropsy evaluation of a representative sample of the population. At the end of each experiment, all birds were evaluated for BCO. A high incidence of BCO was found, even in birds with no external signs of lameness. However, the incidence of BCO was not correlated with the intestinal challenge. Conclusions: under the conditions used in these studies, a treatment that is associated with severe damage to the intestinal mucosa does not change the incidence of BCO in broiler chickens.

The premammillary nucleus of the hypothalamus is not necessary for photoperiodic timekeeping in female turkeys (Meleagris gallopavo).

In birds, seasonal reproduction is regulated by day length, with long days in the spring activating the hypothalamic-pituitary-gonadal axis and reproductive behaviors. The photoreceptors mediating this process remain unknown, but recently, the premammillary nucleus (PMM) of the hypothalamus has been implicated as the site of photoperiodic signaling in turkeys. We performed electrolytic lesions of the PMM to elucidate its role in the photoactivation and maintenance of egg production in female turkeys. Our results show that ablation of the PMM does not alter the normal lay cycle. No differences were found between lesioned birds and sham controls in the latency to lay following photostimulation, nor in subsequent egg production over a period of 29 weeks. No differences in the incidence of gonadal regression were found, indicating that the PMM is not essential for the termination of breeding. We conclude that any role of the PMM in photoperiodic regulation, if it exists, is redundant with other components of the system.

Photic resetting of the circadian clock is correlated with photic habitat in Anolis lizards.

Circadian rhythms are regulated by an internal clock, which is itself synchronized to environmental cues such as light and temperature. It is widely assumed that the circadian system is adapted to local cues, which vary enormously across habitats, yet the comparative data necessary for testing this idea are lacking. We examined photic and thermal resetting of the circadian clock in five species of Anolis lizards whose microhabitats differ in the amounts of sun and shade. The primary circadian oscillator in Anolis is the pineal gland, which produces the hormone melatonin. A flow-through culture system was employed to measure rhythmic melatonin output from individually cultured pineal glands. All species showed temperature-compensated circadian rhythms of pineal melatonin. Light caused significant phase delays of the melatonin rhythm, and this effect varied among species. Controlling for phylogenetic differences, the results indicate that the pineal glands of shade-dwelling species are more sensitive to photic resetting than species living in more brightly illuminated habitats. The differences were not due to variation in free-running period, but may be due to variation in oscillator phase and/or robustness. Surprisingly, thermal resetting was not statistically significant. Overall, the results suggest that the Anolis circadian system is adapted to photic habitat.

Photic induction of locomotor activity is correlated with photic habitat in Anolis lizards.

A variety of ecologically important behaviors, including circadian rhythms and seasonal reproduction, are influenced by non-visual responses to light, yet very little is known about the relationship between photic habitat and non-visual photoreception. Puerto Rican Anolis lizards have diverged into multiple photic niches, making them a good model for non-visual photosensory ecology. We investigated the photic induction of locomotor activity, a non-visual response to light, in four species of Anolis comprising two pairs of closely related, ecomorphologically similar species whose microhabitats differ in solar irradiance. We developed a device for continuous, automated detection and recording of anole locomotor activity, and used it to characterize activity under 12:12 h light-dark cycles. Next, we administered a series of 2-h light pulses during the dark period of the light-dark cycle and measured the increase in locomotor activity relative to baseline dark activity. Five different irradiances (ranging from very dim to daytime levels) were given to each individual lizard on separate nights. As expected, light caused an irradiance-dependent increase in locomotor activity in all four species. The responses at the highest irradiances were significantly greater in species occupying relatively more shaded habitats, suggesting that non-visual photoreception may be adapted to habitat light in Anolis lizards.

The effect of light on melatonin secretion in the cultured pineal glands of Anolis lizards.

Melatonin, a hormone produced by the pineal gland, is important for regulating circadian rhythms in many animals. Light at night causes an acute suppression of melatonin in nearly all vertebrate species. A previous study found that light failed to suppress melatonin in the lizard Anolis carolinensis. This is a surprising result given that the Anolis pineal gland is intrinsically photosensitive, is a key pacemaker controlling locomotor activity, and can be directly entrained to a light-dark cycle. To find out if the lack of photic suppression is widespread in the Anolis genus, we investigated the acute effects of light on melatonin secretion in five different species of Anolis using flow-through tissue culture. We administered a two-hour pulse of bright light to isolated pineal glands during the night. The results show photic suppression of melatonin in all five Anolis species, but the suppression is weak relative to that seen in other vertebrates. Moreover, Anolis species differ in the magnitude of the effect. These findings are discussed in the context of vertebrate pineal evolution and the ecology of Anolis lizards. Given their extensive phylogenetic and ecological divergence, Anolis lizards provide a promising system for investigating the ecology and evolution of circadian organization.

Modulation of metabolic and clock gene mRNA rhythms by pineal and retinal circadian oscillators.

Avian circadian organization involves interactions between three neural pacemakers: the suprachiasmatic nuclei (SCN), pineal, and retina. Each of these structures is linked within a neuroendocrine loop to influence downstream processes and peripheral oscillations. However, the contribution of each structure to drive or synchronize peripheral oscillators or circadian outputs in avian species is largely unknown. To explore these interactions in the chick, we measured 2-deoxy[(14)C]-glucose (2DG) uptake and mRNA expression of the chick clock genes bmal1, cry1, and per3 in three brain areas and in two peripheral organs in chicks that underwent pinealectomy, enucleation, or sham surgery. We found that 2DG uptake rhythms damp under constant darkness in intact animals, while clock gene mRNA levels continue to cycle, demonstrating that metabolic rhythms are not directly driven by clock gene transcription. Moreover, 2DG rhythms are not phase-locked to rhythms of clock gene mRNA. However, pinealectomy and enucleation had similar disruptive effects on both metabolic and clock gene rhythms, suggesting that both of these oscillators act similarly to reinforce molecular and physiological rhythms in the chicken. Finally, we show that the relative phasing of at least one clock gene, cry1, varies between central and peripheral oscillators in a tissue specific manner. These data point to a complex, differential orchestration of central and peripheral oscillators in the chick, and, importantly, indicate a disconnect between canonical clock gene regulation and circadian control of metabolism.

Genealogical concordance and the specific status of Peromyscus sejugis.

Peromyscus sejugis, a peripheral isolate of Peromyscus maniculatus, is a threatened taxon endemic to 2 small islands in the Sea of Cortés. Although its insularity makes the specific recognition of P. sejugis inherently problematic, resolution of this problem has important conservation implications. To evaluate the specific validity and evolutionary history of P. sejugis, we compared sequence variation (ND3/ND4L/ND4) in mtDNA for both island populations of P. sejugis with that for 8 populations of P. maniculatus from mainland Baja California. Each island population of P. sejugis had a single haplotype (0.7% sequence divergence), whereas 11 different haplotypes (mean sequence divergence = 0.68%) were obtained for the populations of P. maniculatus. The mean sequence divergence between the populations of the 2 species was 2.0%. Nested clade analysis supports the conclusion that P. sejugis is an insular isolate of P. maniculatus from mainland Baja California. Although our analysis confirms a low level of mtDNA divergence between P. sejugis and P. maniculatus from Baja California, the genealogical concordance of morphological, chromosomal, microsatellite, and mtDNA haplotype distinctiveness supports the conclusion that the 2 island populations of P. sejugis constitute independent evolutionarily significant units and together represent a phylogenetic species distinct from the P. maniculatus from Baja California.