Protein electrophoresis is recognized as a reliable diagnostic tool for birds even though results... more Protein electrophoresis is recognized as a reliable diagnostic tool for birds even though results are seldom pathognomonic. Unfortunately, this technique is underused in avian medicine because many factors interfere with electrophoresis patterns; hemolysis is one of these factors and is often associated with improper specimen handling. In human laboratory medicine, hemolysis is a known interference factor that can lead to erroneous results. Published data on the influence of hemolysis on protein electrophoresis in birds is currently restricted to a single study in Psittacidae. The aim of this study was to further investigate this effect and to analyze potential interspecific differences. Blood samples were drawn from 28 Black Kites (Milvus migrans) and 19 Bar-headed Geese (Anser indicus) and separated into two aliquots. One aliquot was dipped into liquid nitrogen for 5 sec in order to cause freeze-thawing hemolysis before centrifugation. Total plasma protein concentration, plasma hemoglobin concentration, and plasma protein electrophoresis patterns were determined for both hemolyzed and nonhemolyzed samples. In both species, hemolysis resulted in falsely elevated total plasma protein concentration. In Bar-headed Geese, hemolysis caused a rise in the gamma fraction. In Black Kites, this rise involved not only the gamma fraction but also the beta fraction, stressing the potential for species-related differences. In both species, the effects of hemolysis mimicked a chronic inflammatory condition with resulting antigenic stimulation.
The evolution of multiple sexual traits remains controversial and poorly understood in evolutiona... more The evolution of multiple sexual traits remains controversial and poorly understood in evolutionary biology. In many bird species, males exhibit complex courtships involving feather ornaments and behavioral display. Multiple traits may convey information on the genetic and phenotypic quality of males. In particular, fixed characters, such as feather ornaments (produced once a year during the annual molt in many bird species) might convey information about past male condition (at the time of trait development); whereas flexible traits such as behavioral displays should be sensitive to present condition. Here we show that both flexible behavioral displays and fixed feather ornaments of peacocks, used by females to choose a mate, honestly reflect health status. Correlative data showed that peacocks with high display rate (the number of behavioral displays per hour) and a large number of tail eyespots had low levels of circulating heterophils, suggesting better health status. Experimental activation of the immune system, through LPS injection, significantly reduced display rate compared to a control group. However, the sensitivity of a male display rate to the immune challenge Communicated by M. Leonard A. Loyau ( ) was dependent on the number of tail eyespots: males with higher number of tail eyespots were better able to cope with the experimental immune challenge, and maintained their display rate at levels closer to the levels performed before the immune activation. These results are consistent with the idea that multiple signaling might enhance information reliability.
Studies of the movements and home-ranges of houbara bustards (Chlamydotis undulata undulata) show... more Studies of the movements and home-ranges of houbara bustards (Chlamydotis undulata undulata) showed sexual and seasonal differences in the use of space, with a polygynous mating system similar to an 'exploded-lek' or a 'resource-defence-polygyny', that remains undefined. We used the arthropod biomass as an index of the trophic quality of six defined habitats and we radio-tracked 7 females and 13 males to test whether sexual and seasonal variations in habitat use were related to resource availability, and to verify if critical resources for breeding females were monopolised by males. We analysed habitat selection in both sexes separately. We used the habitat type composition of buffer zones around radio-locations to study annual and seasonal habitat selection and to identify preferred habitats, using the chi-square goodness-of-fit test. Habitat use between sexes and between seasons were compared using MANOVA based on log-ratios of habitat proportions. During the year, and in each season, both sexes appeared to be significantly selective for habitats in comparison to their availability. But males avoided esparto grass, while females used all habitats. Habitat use differed between sexes in the breeding season, but not in the non-breeding season. In spring, when food resources were abundant and uniformly distributed in space, males preferred 'temporarily flooded areas' and females preferred 'reg with tall perennials' that offered both food and cover for brooding. Critical resources were not monopolised by males and the mating system fulfilled the definition of the 'exploded-lek'. Leks are key sites for reproduction and should be considered as priority areas in further conservation plans.
13C & 15N isotope fractionations in Stuthio camelus eggshells: a tool for ecological and environn... more 13C & 15N isotope fractionations in Stuthio camelus eggshells: a tool for ecological and environnemental reconstructions
Although plasma protein electrophoresis is an invaluable diagnostic tool in avian medicine, high ... more Although plasma protein electrophoresis is an invaluable diagnostic tool in avian medicine, high inter-taxonomic variations sometimes make their interpretation difficult for practitioners. The purpose of this study was to improve global understanding of avian plasma electrophoresis patterns by locating the fibrinogen and albumin, and by comparing migration distances of the latter in different avian species. This study was conducted on 80 birds from five different species: 15 peafowls (Pavo cristatus), 18 bar-headed geese (Anser indicus), 12 rock doves (Columba livia), 21 black kites (Milvus migrans) and 14 orange-winged parrots (Amazona amazonica). Protein electrophoresis was performed on both plasma and serum. For each bird, significant differences between fraction concentrations were taken into account to determine the position of the fibrinogen peak on both electrophoresis curves and agarose gels in each species. Once the fibrinogen was located, measurements were performed on the...
Understanding whether the sperm of older males has a diminished capacity to produce successful of... more Understanding whether the sperm of older males has a diminished capacity to produce successful offspring is a key challenge in evolutionary biology. We investigate this issue using 10 years of reproductive data on captive long-lived houbara bustards (Chlamydotis undulata), where the use of artificial insemination techniques means parents can only influence offspring quality via their gametes. Here we show that paternal aging reduces both the likelihood that eggs hatch and the rate at which chicks grow, with older males producing the lightest offspring after the first month. Surprisingly, this cost of paternal aging on offspring development is of a similar scale to that associated with maternal aging. Fitting with predictions on germline aging, the sperm of immature males produce the fastest growing offspring. Our findings thus indicate that any good genes benefit that might be offered by older 'proven' males will be eroded by aging of their germline DNA.
Background: The ''good genes'' theory of sexual selection postulates that females choose mates th... more Background: The ''good genes'' theory of sexual selection postulates that females choose mates that will improve their offspring's fitness through the inheritance of paternal genes. In spite of the attention that this hypothesis has given rise to, the empirical evidence remains sparse, mostly because of the difficulties of controlling for the many environmental factors that may covary with both the paternal phenotype and offspring fitness. Here, we tested the hypothesis that offspring sired by males of a preferred phenotype should have better survival in an endangered bird, the houbara bustard (Chlamydotis undulata undulata).
Despite recent efforts to develop the science of reintroduction biology, there is still no genera... more Despite recent efforts to develop the science of reintroduction biology, there is still no general 24 and broadly accepted definition of reintroduction success. We investigate this issue based on 25 the postulates that (1) successful reintroduction programs should produce viable populations, 26
Finding food resources and maintaining a balanced diet are major concerns for all animals. A comp... more Finding food resources and maintaining a balanced diet are major concerns for all animals. A compromise between neophobia and neophilia is hypothesised to enable animals to enlarge their diet while limiting the risk of poisoning. However, little is known about how primates respond to novel food items and whether their use is socially transmitted. By comparing how four different species of great apes respond to novel food items, we investigated how differences in physiology (digestive tract size and microbial content), habitats (predictability of food availability), and social systems (group size and composition) affect their response toward novelty. We presented two familiar foods, one novel fruit, four novel aromatic plants from herbal medicine, and kaolin to captive chimpanzees (Pan troglodytes), western gorillas (Gorilla gorilla), Bornean orangutans (Pongo pygmaeus) and Sumatran orangutans (Pongo abelii). We recorded smelling, approach-taste delays, ingestion, interindividual observations, and food transfers with continuous sampling. We found that behaviors differed between the apes: chimpanzees were the most cautious species and observed their conspecifics handling the items more frequently than the other apes. Close observations and food transfers were extremely rare in gorillas in comparison to orangutans and chimpanzees. We suggest that a low neophobia level reflects an adaptive response to digestive physiological features in gorillas and to unpredictable food availability in orangutans. Social interactions appeared to be predominant in chimpanzees and in both orangutan Int J Primatol species to overcome food neophobia. They reflect higher social tolerance and more opportunities for social learning and cultural transmission in a feeding context.
Plasma protein electrophoresis is frequently used in birds as a tool for the diagnosis and monito... more Plasma protein electrophoresis is frequently used in birds as a tool for the diagnosis and monitoring of disease. Identification of proteins in individual peaks can help improve our understanding of changes in protein concentration in physiologic and pathologic conditions. The aim of this study was to verify the presence and identity the protein(s) in the prominent alpha-globulin peak of orange-winged parrots (Amazona amazonica), black kites (Milvus migrans), and rock pigeons (Columba livia). Heparinized plasma samples were obtained from 12 birds of each species. Agarose gel electrophoresis and total protein concentration were determined using standard techniques. One plasma sample from each species was then electrophoresed using high-resolution agarose gels to isolate the alpha-globulin band. Gel strips were digested in trypsin and peptides were extracted and analyzed using liquid chromatography with tandem mass spectrometry. De novo sequencing was used to identify the protein based on homology scoring against a protein database. Electrophoresis verified the presence of a single prominent alpha-globulin peak, usually in the alpha(1)-region, that had a median concentration of 9.4 g/L (range, 2.1-11.7 g/L, 21.6% of total protein) in parrots, 12.2 g/L (10.4-13.2 g/L, 35.9%) in kites, and 10.7 g/L (9.0-11.5 g/L, 40.0%) in pigeons. Mass spectrometry and sequencing analysis unequivocally identified the protein as a mature circulating form of apolipoprotein A-I (apo A-I) in all 3 species. Apo A-I accounts for the prominent alpha-globulin peak and comprises a major proportion of total protein concentration in diverse avian species. As a high-density lipoprotein and negative acute phase protein with a pivotal role in cholesterol homeostasis, further study is warranted to determine the significance of changes in apo A-I concentration in avian electrophoretograms.
In order to improve the genetic management of bird species within the European Endangered Program... more In order to improve the genetic management of bird species within the European Endangered Programs (EEP), a research project on artiĀ®cial insemination and cryopreservation of Galliformes semen has been developed. The aim of the program is to create a sperm cryobank for threatened bird species. During this study, semen was collected from 17 pheasant species and speciĀ®c characteristics of ejaculates were analyzed (volume, sperm concentration, motility, pH). ArtiĀ®cial insemination with fresh semen was performed in nine species and with frozen semen in eight species. Inseminations with frozen and thawed semen were made in 17 species. Viability of fresh and frozen semen was assessed in vitro using double stains, eosin and nigrosin. The effect of pH (7Ā±8.5) on viability of fresh and frozen/thawed spermatozoa was also studied. Chicks hatched in eight and three species after insemination with fresh and frozen/thawed semen, respectively. Species varied widely in semen viability: 1Ā±30% of spermatozoa survived freezing and thawing. There was a negative correlation between the viability of frozen spermatozoa and semen pH. In our experimental conditions, the pH of diluents had no effect on semen viability. However, semen with the highest pH had the lowest quality after freezing and thawing. These experiments demonstrated the feasibility of using a very simple and inexpensive method to achieve artiĀ®cial insemination and cryopreservation of semen in endangered pheasant species. #
Semen cryopreservation is very important for the ex situ management of genetic diversity in birds... more Semen cryopreservation is very important for the ex situ management of genetic diversity in birds but it is rarely used. This is partly because of the highly variable success rates, and this emphasizes the need for predictors of semen freezability. This study evaluated the ability of semen quality tests to predict the success rates of semen cryopreservation in chickens and the relationships between each test. Individual variations of in vitro quality tests of semen were compared to the fertility obtained with fresh and cryopreserved semen. The in vitro semen quality tests represented viability, integrity, motility (percentage of viable and morphologically normal cells (PVN); mass motility (MMOT) and different motion parameters including percentage of motile spermatozoa (PMOT)) and biophysical tests (OSM, resistance to osmotic stress; membrane fluidity (FLUID)). Different in vitro tests were significantly correlated between each other for fresh (MMOT, PVN and FLUID, many criteria of objective motility) and cryopreserved semen (MMOT, different objective motility parameters, PVN). Fertility was significantly correlated with PVN for fresh semen and PVN and different objective motility criteria for cryopreserved semen. Membrane fluidity, followed by PVN, PMOT and MMOT, measured on fresh semen samples was positively correlated with fertility obtained with cryopreserved semen. The combination of the first three tests explained 85% of the variability of fertility observed with cryopreserved semen. In conclusion, we showed that different in vitro tests of semen quality are of predictive value for the success rate of semen cryopreservation in the chicken, the most accurate being membrane fluidity.
Protein electrophoresis is recognized as a reliable diagnostic tool for birds even though results... more Protein electrophoresis is recognized as a reliable diagnostic tool for birds even though results are seldom pathognomonic. Unfortunately, this technique is underused in avian medicine because many factors interfere with electrophoresis patterns; hemolysis is one of these factors and is often associated with improper specimen handling. In human laboratory medicine, hemolysis is a known interference factor that can lead to erroneous results. Published data on the influence of hemolysis on protein electrophoresis in birds is currently restricted to a single study in Psittacidae. The aim of this study was to further investigate this effect and to analyze potential interspecific differences. Blood samples were drawn from 28 Black Kites (Milvus migrans) and 19 Bar-headed Geese (Anser indicus) and separated into two aliquots. One aliquot was dipped into liquid nitrogen for 5 sec in order to cause freeze-thawing hemolysis before centrifugation. Total plasma protein concentration, plasma hemoglobin concentration, and plasma protein electrophoresis patterns were determined for both hemolyzed and nonhemolyzed samples. In both species, hemolysis resulted in falsely elevated total plasma protein concentration. In Bar-headed Geese, hemolysis caused a rise in the gamma fraction. In Black Kites, this rise involved not only the gamma fraction but also the beta fraction, stressing the potential for species-related differences. In both species, the effects of hemolysis mimicked a chronic inflammatory condition with resulting antigenic stimulation.
Plasma protein electrophoresis is recognized as a reliable diagnostic tool in avian medicine; how... more Plasma protein electrophoresis is recognized as a reliable diagnostic tool in avian medicine; however, the influence of circannual phenomena such as molt on protein electrophoregrams is poorly documented. The molt is a period of heavy hormonal and metabolic change in birds. The purpose of this study was to investigate the effects of molt on total protein concentration and electrophoresis patterns in birds. Blood samples were taken from 19 Bar-headed Geese (Anser indicus) from mid-May to mid-August, at 15-day intervals. At the same time, molting stage of each bird was recorded. Total protein concentrations were measured and plasma agarose gel electrophoresis was performed on these samples. The Bar-headed Goose was chosen as a model, because they molt over a very short period. The total protein concentration and albumin, alpha-2, beta, and gamma fractions were at their minimum values during molt, whereas the prealbumin and alpha-1 fractions rose to their maximum levels. This study provides baseline information relevant to changes occurring in avian proteinograms throughout the molt. The increase in prealbumin and alpha-1 fractions may be related to an increase in plasma thyroid hormones during molt. The decrease observed in albumin, alpha-2, beta, and gamma fractions may be related to protein and energy shifts toward feather growth, as well as to an expansion of the circulatory system located around the feather follicles with secondary dilutional effects on protein fractions. From a clinical point of view, the observed changes associated with molting were less significant than initially expected, and would not likely results in incorrect diagnoses based on interpretation of the protein electrophoretic patterns.
Studies of the movements and home-ranges of houbara bustards (Chlamydotis undulata undulata) show... more Studies of the movements and home-ranges of houbara bustards (Chlamydotis undulata undulata) showed sexual and seasonal differences in the use of space, with a polygynous mating system similar to an 'exploded-lek' or a 'resource-defence-polygyny', that remains undefined. We used the arthropod biomass as an index of the trophic quality of six defined habitats and we radio-tracked 7 females and 13 males to test whether sexual and seasonal variations in habitat use were related to resource availability, and to verify if critical resources for breeding females were monopolised by males. We analysed habitat selection in both sexes separately. We used the habitat type composition of buffer zones around radio-locations to study annual and seasonal habitat selection and to identify preferred habitats, using the chi-square goodness-of-fit test. Habitat use between sexes and between seasons were compared using MANOVA based on log-ratios of habitat proportions. During the year, and in each season, both sexes appeared to be significantly selective for habitats in comparison to their availability. But males avoided esparto grass, while females used all habitats. Habitat use differed between sexes in the breeding season, but not in the non-breeding season. In spring, when food resources were abundant and uniformly distributed in space, males preferred 'temporarily flooded areas' and females preferred 'reg with tall perennials' that offered both food and cover for brooding. Critical resources were not monopolised by males and the mating system fulfilled the definition of the 'exploded-lek'. Leks are key sites for reproduction and should be considered as priority areas in further conservation plans.
Protein electrophoresis is recognized as a reliable diagnostic tool for birds even though results... more Protein electrophoresis is recognized as a reliable diagnostic tool for birds even though results are seldom pathognomonic. Unfortunately, this technique is underused in avian medicine because many factors interfere with electrophoresis patterns; hemolysis is one of these factors and is often associated with improper specimen handling. In human laboratory medicine, hemolysis is a known interference factor that can lead to erroneous results. Published data on the influence of hemolysis on protein electrophoresis in birds is currently restricted to a single study in Psittacidae. The aim of this study was to further investigate this effect and to analyze potential interspecific differences. Blood samples were drawn from 28 Black Kites (Milvus migrans) and 19 Bar-headed Geese (Anser indicus) and separated into two aliquots. One aliquot was dipped into liquid nitrogen for 5 sec in order to cause freeze-thawing hemolysis before centrifugation. Total plasma protein concentration, plasma hemoglobin concentration, and plasma protein electrophoresis patterns were determined for both hemolyzed and nonhemolyzed samples. In both species, hemolysis resulted in falsely elevated total plasma protein concentration. In Bar-headed Geese, hemolysis caused a rise in the gamma fraction. In Black Kites, this rise involved not only the gamma fraction but also the beta fraction, stressing the potential for species-related differences. In both species, the effects of hemolysis mimicked a chronic inflammatory condition with resulting antigenic stimulation.
The evolution of multiple sexual traits remains controversial and poorly understood in evolutiona... more The evolution of multiple sexual traits remains controversial and poorly understood in evolutionary biology. In many bird species, males exhibit complex courtships involving feather ornaments and behavioral display. Multiple traits may convey information on the genetic and phenotypic quality of males. In particular, fixed characters, such as feather ornaments (produced once a year during the annual molt in many bird species) might convey information about past male condition (at the time of trait development); whereas flexible traits such as behavioral displays should be sensitive to present condition. Here we show that both flexible behavioral displays and fixed feather ornaments of peacocks, used by females to choose a mate, honestly reflect health status. Correlative data showed that peacocks with high display rate (the number of behavioral displays per hour) and a large number of tail eyespots had low levels of circulating heterophils, suggesting better health status. Experimental activation of the immune system, through LPS injection, significantly reduced display rate compared to a control group. However, the sensitivity of a male display rate to the immune challenge Communicated by M. Leonard A. Loyau ( ) was dependent on the number of tail eyespots: males with higher number of tail eyespots were better able to cope with the experimental immune challenge, and maintained their display rate at levels closer to the levels performed before the immune activation. These results are consistent with the idea that multiple signaling might enhance information reliability.
Studies of the movements and home-ranges of houbara bustards (Chlamydotis undulata undulata) show... more Studies of the movements and home-ranges of houbara bustards (Chlamydotis undulata undulata) showed sexual and seasonal differences in the use of space, with a polygynous mating system similar to an 'exploded-lek' or a 'resource-defence-polygyny', that remains undefined. We used the arthropod biomass as an index of the trophic quality of six defined habitats and we radio-tracked 7 females and 13 males to test whether sexual and seasonal variations in habitat use were related to resource availability, and to verify if critical resources for breeding females were monopolised by males. We analysed habitat selection in both sexes separately. We used the habitat type composition of buffer zones around radio-locations to study annual and seasonal habitat selection and to identify preferred habitats, using the chi-square goodness-of-fit test. Habitat use between sexes and between seasons were compared using MANOVA based on log-ratios of habitat proportions. During the year, and in each season, both sexes appeared to be significantly selective for habitats in comparison to their availability. But males avoided esparto grass, while females used all habitats. Habitat use differed between sexes in the breeding season, but not in the non-breeding season. In spring, when food resources were abundant and uniformly distributed in space, males preferred 'temporarily flooded areas' and females preferred 'reg with tall perennials' that offered both food and cover for brooding. Critical resources were not monopolised by males and the mating system fulfilled the definition of the 'exploded-lek'. Leks are key sites for reproduction and should be considered as priority areas in further conservation plans.
13C & 15N isotope fractionations in Stuthio camelus eggshells: a tool for ecological and environn... more 13C & 15N isotope fractionations in Stuthio camelus eggshells: a tool for ecological and environnemental reconstructions
Although plasma protein electrophoresis is an invaluable diagnostic tool in avian medicine, high ... more Although plasma protein electrophoresis is an invaluable diagnostic tool in avian medicine, high inter-taxonomic variations sometimes make their interpretation difficult for practitioners. The purpose of this study was to improve global understanding of avian plasma electrophoresis patterns by locating the fibrinogen and albumin, and by comparing migration distances of the latter in different avian species. This study was conducted on 80 birds from five different species: 15 peafowls (Pavo cristatus), 18 bar-headed geese (Anser indicus), 12 rock doves (Columba livia), 21 black kites (Milvus migrans) and 14 orange-winged parrots (Amazona amazonica). Protein electrophoresis was performed on both plasma and serum. For each bird, significant differences between fraction concentrations were taken into account to determine the position of the fibrinogen peak on both electrophoresis curves and agarose gels in each species. Once the fibrinogen was located, measurements were performed on the...
Understanding whether the sperm of older males has a diminished capacity to produce successful of... more Understanding whether the sperm of older males has a diminished capacity to produce successful offspring is a key challenge in evolutionary biology. We investigate this issue using 10 years of reproductive data on captive long-lived houbara bustards (Chlamydotis undulata), where the use of artificial insemination techniques means parents can only influence offspring quality via their gametes. Here we show that paternal aging reduces both the likelihood that eggs hatch and the rate at which chicks grow, with older males producing the lightest offspring after the first month. Surprisingly, this cost of paternal aging on offspring development is of a similar scale to that associated with maternal aging. Fitting with predictions on germline aging, the sperm of immature males produce the fastest growing offspring. Our findings thus indicate that any good genes benefit that might be offered by older 'proven' males will be eroded by aging of their germline DNA.
Background: The ''good genes'' theory of sexual selection postulates that females choose mates th... more Background: The ''good genes'' theory of sexual selection postulates that females choose mates that will improve their offspring's fitness through the inheritance of paternal genes. In spite of the attention that this hypothesis has given rise to, the empirical evidence remains sparse, mostly because of the difficulties of controlling for the many environmental factors that may covary with both the paternal phenotype and offspring fitness. Here, we tested the hypothesis that offspring sired by males of a preferred phenotype should have better survival in an endangered bird, the houbara bustard (Chlamydotis undulata undulata).
Despite recent efforts to develop the science of reintroduction biology, there is still no genera... more Despite recent efforts to develop the science of reintroduction biology, there is still no general 24 and broadly accepted definition of reintroduction success. We investigate this issue based on 25 the postulates that (1) successful reintroduction programs should produce viable populations, 26
Finding food resources and maintaining a balanced diet are major concerns for all animals. A comp... more Finding food resources and maintaining a balanced diet are major concerns for all animals. A compromise between neophobia and neophilia is hypothesised to enable animals to enlarge their diet while limiting the risk of poisoning. However, little is known about how primates respond to novel food items and whether their use is socially transmitted. By comparing how four different species of great apes respond to novel food items, we investigated how differences in physiology (digestive tract size and microbial content), habitats (predictability of food availability), and social systems (group size and composition) affect their response toward novelty. We presented two familiar foods, one novel fruit, four novel aromatic plants from herbal medicine, and kaolin to captive chimpanzees (Pan troglodytes), western gorillas (Gorilla gorilla), Bornean orangutans (Pongo pygmaeus) and Sumatran orangutans (Pongo abelii). We recorded smelling, approach-taste delays, ingestion, interindividual observations, and food transfers with continuous sampling. We found that behaviors differed between the apes: chimpanzees were the most cautious species and observed their conspecifics handling the items more frequently than the other apes. Close observations and food transfers were extremely rare in gorillas in comparison to orangutans and chimpanzees. We suggest that a low neophobia level reflects an adaptive response to digestive physiological features in gorillas and to unpredictable food availability in orangutans. Social interactions appeared to be predominant in chimpanzees and in both orangutan Int J Primatol species to overcome food neophobia. They reflect higher social tolerance and more opportunities for social learning and cultural transmission in a feeding context.
Plasma protein electrophoresis is frequently used in birds as a tool for the diagnosis and monito... more Plasma protein electrophoresis is frequently used in birds as a tool for the diagnosis and monitoring of disease. Identification of proteins in individual peaks can help improve our understanding of changes in protein concentration in physiologic and pathologic conditions. The aim of this study was to verify the presence and identity the protein(s) in the prominent alpha-globulin peak of orange-winged parrots (Amazona amazonica), black kites (Milvus migrans), and rock pigeons (Columba livia). Heparinized plasma samples were obtained from 12 birds of each species. Agarose gel electrophoresis and total protein concentration were determined using standard techniques. One plasma sample from each species was then electrophoresed using high-resolution agarose gels to isolate the alpha-globulin band. Gel strips were digested in trypsin and peptides were extracted and analyzed using liquid chromatography with tandem mass spectrometry. De novo sequencing was used to identify the protein based on homology scoring against a protein database. Electrophoresis verified the presence of a single prominent alpha-globulin peak, usually in the alpha(1)-region, that had a median concentration of 9.4 g/L (range, 2.1-11.7 g/L, 21.6% of total protein) in parrots, 12.2 g/L (10.4-13.2 g/L, 35.9%) in kites, and 10.7 g/L (9.0-11.5 g/L, 40.0%) in pigeons. Mass spectrometry and sequencing analysis unequivocally identified the protein as a mature circulating form of apolipoprotein A-I (apo A-I) in all 3 species. Apo A-I accounts for the prominent alpha-globulin peak and comprises a major proportion of total protein concentration in diverse avian species. As a high-density lipoprotein and negative acute phase protein with a pivotal role in cholesterol homeostasis, further study is warranted to determine the significance of changes in apo A-I concentration in avian electrophoretograms.
In order to improve the genetic management of bird species within the European Endangered Program... more In order to improve the genetic management of bird species within the European Endangered Programs (EEP), a research project on artiĀ®cial insemination and cryopreservation of Galliformes semen has been developed. The aim of the program is to create a sperm cryobank for threatened bird species. During this study, semen was collected from 17 pheasant species and speciĀ®c characteristics of ejaculates were analyzed (volume, sperm concentration, motility, pH). ArtiĀ®cial insemination with fresh semen was performed in nine species and with frozen semen in eight species. Inseminations with frozen and thawed semen were made in 17 species. Viability of fresh and frozen semen was assessed in vitro using double stains, eosin and nigrosin. The effect of pH (7Ā±8.5) on viability of fresh and frozen/thawed spermatozoa was also studied. Chicks hatched in eight and three species after insemination with fresh and frozen/thawed semen, respectively. Species varied widely in semen viability: 1Ā±30% of spermatozoa survived freezing and thawing. There was a negative correlation between the viability of frozen spermatozoa and semen pH. In our experimental conditions, the pH of diluents had no effect on semen viability. However, semen with the highest pH had the lowest quality after freezing and thawing. These experiments demonstrated the feasibility of using a very simple and inexpensive method to achieve artiĀ®cial insemination and cryopreservation of semen in endangered pheasant species. #
Semen cryopreservation is very important for the ex situ management of genetic diversity in birds... more Semen cryopreservation is very important for the ex situ management of genetic diversity in birds but it is rarely used. This is partly because of the highly variable success rates, and this emphasizes the need for predictors of semen freezability. This study evaluated the ability of semen quality tests to predict the success rates of semen cryopreservation in chickens and the relationships between each test. Individual variations of in vitro quality tests of semen were compared to the fertility obtained with fresh and cryopreserved semen. The in vitro semen quality tests represented viability, integrity, motility (percentage of viable and morphologically normal cells (PVN); mass motility (MMOT) and different motion parameters including percentage of motile spermatozoa (PMOT)) and biophysical tests (OSM, resistance to osmotic stress; membrane fluidity (FLUID)). Different in vitro tests were significantly correlated between each other for fresh (MMOT, PVN and FLUID, many criteria of objective motility) and cryopreserved semen (MMOT, different objective motility parameters, PVN). Fertility was significantly correlated with PVN for fresh semen and PVN and different objective motility criteria for cryopreserved semen. Membrane fluidity, followed by PVN, PMOT and MMOT, measured on fresh semen samples was positively correlated with fertility obtained with cryopreserved semen. The combination of the first three tests explained 85% of the variability of fertility observed with cryopreserved semen. In conclusion, we showed that different in vitro tests of semen quality are of predictive value for the success rate of semen cryopreservation in the chicken, the most accurate being membrane fluidity.
Protein electrophoresis is recognized as a reliable diagnostic tool for birds even though results... more Protein electrophoresis is recognized as a reliable diagnostic tool for birds even though results are seldom pathognomonic. Unfortunately, this technique is underused in avian medicine because many factors interfere with electrophoresis patterns; hemolysis is one of these factors and is often associated with improper specimen handling. In human laboratory medicine, hemolysis is a known interference factor that can lead to erroneous results. Published data on the influence of hemolysis on protein electrophoresis in birds is currently restricted to a single study in Psittacidae. The aim of this study was to further investigate this effect and to analyze potential interspecific differences. Blood samples were drawn from 28 Black Kites (Milvus migrans) and 19 Bar-headed Geese (Anser indicus) and separated into two aliquots. One aliquot was dipped into liquid nitrogen for 5 sec in order to cause freeze-thawing hemolysis before centrifugation. Total plasma protein concentration, plasma hemoglobin concentration, and plasma protein electrophoresis patterns were determined for both hemolyzed and nonhemolyzed samples. In both species, hemolysis resulted in falsely elevated total plasma protein concentration. In Bar-headed Geese, hemolysis caused a rise in the gamma fraction. In Black Kites, this rise involved not only the gamma fraction but also the beta fraction, stressing the potential for species-related differences. In both species, the effects of hemolysis mimicked a chronic inflammatory condition with resulting antigenic stimulation.
Plasma protein electrophoresis is recognized as a reliable diagnostic tool in avian medicine; how... more Plasma protein electrophoresis is recognized as a reliable diagnostic tool in avian medicine; however, the influence of circannual phenomena such as molt on protein electrophoregrams is poorly documented. The molt is a period of heavy hormonal and metabolic change in birds. The purpose of this study was to investigate the effects of molt on total protein concentration and electrophoresis patterns in birds. Blood samples were taken from 19 Bar-headed Geese (Anser indicus) from mid-May to mid-August, at 15-day intervals. At the same time, molting stage of each bird was recorded. Total protein concentrations were measured and plasma agarose gel electrophoresis was performed on these samples. The Bar-headed Goose was chosen as a model, because they molt over a very short period. The total protein concentration and albumin, alpha-2, beta, and gamma fractions were at their minimum values during molt, whereas the prealbumin and alpha-1 fractions rose to their maximum levels. This study provides baseline information relevant to changes occurring in avian proteinograms throughout the molt. The increase in prealbumin and alpha-1 fractions may be related to an increase in plasma thyroid hormones during molt. The decrease observed in albumin, alpha-2, beta, and gamma fractions may be related to protein and energy shifts toward feather growth, as well as to an expansion of the circulatory system located around the feather follicles with secondary dilutional effects on protein fractions. From a clinical point of view, the observed changes associated with molting were less significant than initially expected, and would not likely results in incorrect diagnoses based on interpretation of the protein electrophoretic patterns.
Studies of the movements and home-ranges of houbara bustards (Chlamydotis undulata undulata) show... more Studies of the movements and home-ranges of houbara bustards (Chlamydotis undulata undulata) showed sexual and seasonal differences in the use of space, with a polygynous mating system similar to an 'exploded-lek' or a 'resource-defence-polygyny', that remains undefined. We used the arthropod biomass as an index of the trophic quality of six defined habitats and we radio-tracked 7 females and 13 males to test whether sexual and seasonal variations in habitat use were related to resource availability, and to verify if critical resources for breeding females were monopolised by males. We analysed habitat selection in both sexes separately. We used the habitat type composition of buffer zones around radio-locations to study annual and seasonal habitat selection and to identify preferred habitats, using the chi-square goodness-of-fit test. Habitat use between sexes and between seasons were compared using MANOVA based on log-ratios of habitat proportions. During the year, and in each season, both sexes appeared to be significantly selective for habitats in comparison to their availability. But males avoided esparto grass, while females used all habitats. Habitat use differed between sexes in the breeding season, but not in the non-breeding season. In spring, when food resources were abundant and uniformly distributed in space, males preferred 'temporarily flooded areas' and females preferred 'reg with tall perennials' that offered both food and cover for brooding. Critical resources were not monopolised by males and the mating system fulfilled the definition of the 'exploded-lek'. Leks are key sites for reproduction and should be considered as priority areas in further conservation plans.
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Papers by Michel Jalme