Vietnamese chickens: a gate towards Asian genetic diversity.

BACKGROUND: Chickens represent an important animal genetic resource and the conservation of local breeds is an issue for the preservation of this resource. The genetic diversity of a breed is mainly evaluated through its nuclear diversity. However, nuclear genetic diversity does not provide the same information as mitochondrial genetic diversity. For the species Gallus gallus, at least 8 maternal lineages have been identified. While breeds distributed westward from the Indian subcontinent usually share haplotypes from 1 to 2 haplogroups, Southeast Asian breeds exhibit all the haplogroups. The Vietnamese Ha Giang (HG) chicken has been shown to exhibit a very high nuclear diversity but also important rates of admixture with wild relatives. Its geographical position, within one of the chicken domestication centres ranging from Thailand to the Chinese Yunnan province, increases the probability of observing a very high genetic diversity for maternal lineages, and in a way, improving our understanding of the chicken domestication process. RESULTS: A total of 106 sequences from Vietnamese HG chickens were first compared to the sequences of published Chinese breeds. The 25 haplotypes observed in the Vietnamese HG population belonged to six previously published haplogroups which are: A, B, C, D, F and G. On average, breeds from the Chinese Yunnan province carried haplotypes from 4.3 haplogroups. For the HG population, haplogroup diversity is found at both the province and the village level (0.69).The AMOVA results show that genetic diversity occurred within the breeds rather than between breeds or provinces. Regarding the global structure of the mtDNA diversity per population, a characteristic of the HG population was the occurrence of similar pattern distribution as compared to G. gallus spadiceus. However, there was no geographical evidence of gene flow between wild and domestic populations as observed when microsatellites were used. CONCLUSIONS: In contrast to other chicken populations, the HG chicken population showed very high genetic diversity at both the nuclear and mitochondrial levels. Due to its past and recent history, this population accumulates a specific and rich gene pool highlighting its interest and the need for conservation.

Revealing fine scale subpopulation structure in the Vietnamese H'Mong cattle breed for conservation purposes.

BACKGROUND: During the last decades, there has been an acceleration of the loss of domestic animal biodiversity. For conservation purposes, the genetic diversity of the H'Mong cattle, an indigenous local breed was studied. Single-nucleotide polymorphisms (SNP) of the SRY gene and mtDNA D-Loop sequence were analysed to clarify the origin of the breed. The genetic diversity was assessed through genetic data with twenty-five FAO microsatellites, and morphometric data with five body measurements from 408 animals sampled from eight districts of the Ha Giang province. RESULTS: The SRY genes were all of the zebu type. Among the 27 mtDNA haplotypes, 12 haplotypes were of the taurine type and the remaining 15 of the zebu type. This indicates female taurine introgression in the zebu H'Mong. The observed and expected heterozygosity ranged from 0.616 to 0.673 and from 0.681 to 0.729 respectively according to district, with low genetic differentiation (F ST = 0.0076). Multivariate analysis on morphometric and genetic data shows a separation of districts into two groups following a south-west/north-east cline and admixture analysis confirmed the two clusters, but no differentiation of taurine introgression between clusters was observed. A possible admixture with the Yellow cattle breed from a neighbouring province was suggested through genetic data and householder interviews. CONCLUSIONS: In this study we demonstrate the interest of fine-scale sampling for the study of genetic structure of local breeds. Such a study allows avoiding erroneous conservation policies and on the contrary, proposes measures for conserving and limiting crossbreeding between the H'Mong and the Yellow cattle breeds.

Genetic and morphometric characterization of a local Vietnamese swamp buffalo population.

The water buffalo plays a key role in the socio-economy of South-East Asia as it is the main draught power for paddy rice cultivation. While in the Indian subcontinent the water buffalo is the riverine type, in South-East Asia the majority of buffaloes are of the swamp type. In the poor remote northern province of Ha Giang in Vietnam, improvement of the swamp buffalo breed may be one of the best ways to increase sustainability of farming systems. Therefore, analysis of the genetic structure of the province buffalo population is a prerequisite to any conservation or improvement project. A total of 1122 animals were described for 11 body and horn measurements for morphometric characterization. From this sample set, 744 animals were genotyped for 17 microsatellite markers. Also 17 animals from southern provinces of Vietnam were genotyped as a comparative sample. The results showed that genetic diversity as well as inbreeding value in the Ha Giang was high. The F(ST) values within the province and across Vietnam were low indicating that most of the population variation is explained by individual variability. Bayesian clustering analysis did not highlight the presence of subdivided populations. These results are useful for the implementation of a conservation and improvement strategy of the swamp buffalo in order to guarantee the householders' needs for sustainability of the farming system in the Ha Giang province.

How does farmer connectivity influence livestock genetic structure? A case-study in a Vietnamese goat population.

Assessing how genes flow across populations is a key component of conservation genetics. Gene flow in a natural population depends on ecological traits and the local environment, whereas for a livestock population, gene flow is driven by human activities. Spatial organization, relationships between farmers and their husbandry practices will define the farmer's network and so determine farmer connectivity. It is thus assumed that farmer connectivity will affect the genetic structure of their livestock. To test this hypothesis, goats reared by four different ethnic groups in a Vietnamese province were genotyped using 16 microsatellites. A Bayesian approach and spatial multivariate analysis (spatial principal component analysis, sPCA) were used to identify subpopulations and spatial organization. Ethnic group frequencies, husbandry practices and altitude were used to create cost maps that were implemented in a least-cost path approach. Genetic diversity in the Vietnamese goat population was low (0.508) compared to other local Asian breeds. Using a Bayesian approach, three clusters were identified. sPCA confirmed these three clusters and also that the genetic structure showed a significant spatial pattern. The least-cost path analysis showed that genetic differentiation was significantly correlated (0.131-0.207) to ethnic frequencies and husbandry practices. In brief, the spatial pattern observed in the goat population was the result of complex gene flow governed by the spatial distribution of ethnic groups, ethnicity and husbandry practices. In this study, we clearly linked the livestock genetic pattern to farmer connectivity and showed the importance of taking into account spatial information in genetic studies.

Population genetic structure of wild and farmed rusa deer (Cervus timorensis russa) in New-Caledonia inferred from polymorphic microsatellite loci.

Historical records indicate that 12 rusa deer (Cervus timorensis russa) were introduced in New-Caledonia during the 1870s. We used eight polymorphic microsatellite DNA loci to assess the genetic differentiation and diversity of farmed and wild deer populations. Past genetic bottlenecks were detected in both sub-populations, although higher genetic diversity was maintained in farmed populations, probably due to the regular introduction of reproducers from wild populations and from other farms. The genetic structure of farmed and wild populations differed significantly. There was a significant isolation by distance for wild populations, whereas farmed populations were significantly differentiated between farms independently from their geographical proximity. Wild rusa deer consisted of small populations (with effective population sizes ranging between 7 and 19 individuals depending on the methods used), with a low parent-offspring dispersion range (0.20-2.02 km). Genetic tools and direct observations provided congruent estimates of dispersion and population sizes. We discuss the relevance of our results for management purposes.

Genetic analysis of local Vietnamese chickens provides evidence of gene flow from wild to domestic populations.

UNLABELLED: Previous studies suggested that multiple domestication events in South and South-East Asia (Yunnan and surrounding areas) and India have led to the genesis of modern domestic chickens. Ha Giang province is a northern Vietnamese region, where local chickens, such as the H'mong breed, and wild junglefowl coexist. The assumption was made that hybridisation between wild junglefowl and Ha Giang chickens may have occurred and led to the high genetic diversity previously observed. The objectives of this study were i) to clarify the genetic structure of the chicken population within the Ha Giang province and ii) to give evidence of admixture with G. gallus. A large survey of the molecular polymorphism for 18 microsatellite markers was conducted on 1082 chickens from 30 communes of the Ha Giang province (HG chickens). This dataset was combined with a previous dataset of Asian breeds, commercial lines and samples of Red junglefowl from Thailand and Vietnam (Ha Noï). Measurements of genetic diversity were estimated both within-population and between populations, and a step-by-step Bayesian approach was performed on the global data set. RESULTS: The highest value for expected heterozygosity (> 0.60) was found in HG chickens and in the wild junglefowl populations from Thailand. HG chickens exhibited the highest allelic richness (mean A = 2.9). No significant genetic subdivisions of the chicken population within the Ha Giang province were found. As compared to other breeds, HG chickens clustered with wild populations. Furthermore, the neighbornet tree and the Bayesian clustering analysis showed that chickens from 4 communes were closely related to the wild ones and showed an admixture pattern. CONCLUSION: In the absence of any population structuring within the province, the H'mong chicken, identified from its black phenotype, shared a common gene pool with other chickens from the Ha Giang population. The large number of alleles shared exclusively between Ha Giang chickens and junglefowl, as well as the results of a Bayesian clustering analysis, suggest that gene flow has been taking place from junglefowl to Ha Giang chickens.

Microsatellite analysis of genetic diversity of the Vietnamese sika deer (Cervus nippon pseudaxis).

The Vietnamese sika deer (Cervus nippon pseudaxis) is an endangered subspecies of economic and traditional value in Vietnam. Most living individuals are held in traditional farms in central Vietnam, others being found in zoos around the world. Here we study the neutral genetic diversity and population structure of this subspecies using nine microsatellite loci in order to evaluate the consequences of the limited number of individuals from which this population was initiated and of the breeding practices (i.e., possible inbreeding). Two hundred individuals were sampled from several villages. Our data show both evidence for limited local inbreeding and isolation by distance with a mean F(ST) value of 0.02 between villages. This suggests that exchange of animals occurs at a local scale, at a rate such that highly inbred mating is avoided. However, the genetic diversity, with an expected heterozygosity (H(e)) of 0.60 and mean number of alleles (k) of 5.7, was not significantly larger than that estimated from zoo populations of much smaller census size (17 animals sampled; H(e) = 0.65, k = 4.11). Our results also suggest that the Vietnamese population might have experienced a slight bottleneck. However, this population is sufficiently variable to constitute a source of individuals for reintroduction in the wild in Vietnam.

Contagious bovine pleuropneumonia vaccines, historic highlights, present situation and hopes.

Contagious bovine pleuropneumonia (CBPP) is a contagious infection of cattle caused by a mycoplasma, M. mycoides subsp. mycoides SC (MmmSC). It induces lesions of pleuropneumonia in acute cases and the formation of pulmonary "sequestra" in chronic cases. The disease is prevalent mostly in Africa, where it is responsible for high losses, but it has also been sporadically present in Southern Europe until 1999. Vaccination is now prohibited in most countries except in Africa. An empirical "inoculation" procedure was developed as early as 1852 in Europe but it may have been used even earlier in Africa. The inoculation of pleural fluid was performed at the tip of the tail in Europe and on the bridge of the nose in Africa. It conferred good protection but induced a high number of fatal cases. Various inactivated preparations have been tested in the past with inconclusive results leading sometime to some protection and some other time to a sensitisation of the immunised animals. Such preparations have never been used in the field. Attenuated MmmSC strains have been developed in the 1950s and used extensively in the field both in Africa and Australia. The best known vaccine strains are KH3J, T1/44 and T1sr. Vaccination campaigns have succeeded in reducing considerably the CBPP prevalence in these two continents but eradication was achieved in Australia only by switching to strict measures of animal movement control and a stamping-out policy. The search for new CBPP vaccines has become a major issue for African countries that are facing an increase in outbreaks. The rationale for this search is based on a better understanding of the mycoplasma virulence mechanisms that could lead to a targeted attenuation of MmmSC strains. It is also based on a better understanding of the bovine immune response that may be driven to a pathogenic inflammatory response or conversely to a better balanced response leading to protection.

Efficiency of semi-automated fluorescent multiplex PCRs with 11 microsatellite markers for genetic studies of deer populations.

Thirty bovine and eight ovine microsatellite primer pairs were tested on four tropical deer species: Eld's and Swamp deer (highly threatened) and Rusa and Vietnamese Sika deer (economically important). Thirty markers gave an amplified product in all four species (78.9%). The number of polymorphic microsatellite markers varied among the species from 14 in Eld's deer (47%) to 20 in Swamp deer (67%). Among them, 11 microsatellite loci were multiplexed in three polymerase chain reactions (PCRs) and labelled with three different fluorochromes that can be loaded in one gel-lane. To test the efficiency of the multiplex, primary genetic studies (mean number of alleles, expected heterozygosities and Fis values) were carried out on four deer populations. Parentage exclusion probability and probability of identity were computed and discussed on a Swamp deer population. These multiplexes PCRs were also tested on several other deer species and subspecies. The aim of this study is to establish a tool useful for genetic studies of population structure and diversity in four tropical deer species which with few modifications can be applied to other species of the genus Cervus.

A partial african ancestry for the creole cattle populations of the Caribbean.

Seventy-eight cattle samples from three Creole Caribbean islands and one Brazilian breed were analyzed for sequence variation in the hypervariable segment of the mitochondrial DNA control region. Seventy-three samples displayed Bos taurus haplotypes, and five samples exhibited haplotypes that were of Bos indicus ancestry. Phylogenetic analysis revealed that all sampled B. taurus sequences fell into two distinct clusters with separate African and European origins. European sequences were encountered in each population; however, the distribution of African haplotypes was uneven, with the highest proportion of African influence found in the Guadeloupe Creole. The reduced levels of African haplotypic variation within the Caribbean and Brazilian are consistent with prior founder effects. Additionally, genetic variation at three microsatellite loci illustrated African influence uniquely in the Guadeloupe Creole. Collectively, the data suggest that this African influence is, at least in part, attributable to the historical importation of African cattle to the Americas. Furthermore, alleles of B. indicus ancestry were detected at appreciable frequencies in all Caribbean Creole populations and may reflect zebu introgressions from either West Africa or the Indian subcontinent.

Ehrlichia ruminantium major antigenic protein gene (map1) variants are not geographically constrained and show no evidence of having evolved under positive selection pressure.

In a search for tools to distinguish antigenic variants of Ehrlichia ruminantium, we sequenced the major antigenic protein genes (map1 genes) of 21 different isolates and found that the sequence polymorphisms were too great to permit the design of probes which could be used as markers for immunogenicity. Phylogenetic comparison of the 21 deduced MAP1 sequences plus another 9 sequences which had been previously published did not reveal any geographic clustering among the isolates. Maximum likelihood analysis of codon and amino acid changes over the phylogeny provided no statistical evidence that the gene is under positive selection pressure, suggesting that it may not be important for the evasion of host immune responses.

Characterization of 18 new BoLA-DRB3 alleles.

The second exon of the bovine MHC class II DRB3 gene was amplified by polymerase chain reaction (PCR) from DNA samples of 568 zebu Brahman cattle (Bos indicus) from Martinique (French West Indies). Cloning of these PCR products allowed the isolation of both alleles from each animal, which were characterized by the PCR-restriction fragment length polymorphism (RFLP) technique using the restriction enzymes RsaI, BstYI and HaeIII. Four new PCR-RFLP patterns were obtained by digestion with RsaI. These patterns were named 'v', 'w', 'x' and 'y' continuing the accepted nomenclature. Sequencing of each allele allowed the identification of 18 new BoLA-DRB3 exon 2 nucleotide sequences and their deduced amino acid sequences.

Immune responses to Dermatophilus congolensis infections.

Complex mechanisms underly the establishment of dermatophilosis, an exudative and proliferative skin disease of ruminants. This multicomponent system involves the bacterium Dermatophilus congolensis, transmission by various routes including flies, host genetic factors and immunosuppression by Amblyomma variegatum ticks. Here, Nick Ambrose and colleagues summarize recent evidence for an association between A. variegatum and severe chronic dermatophilosis in cattle. Breed-based differences in resistance to dermatophilosis are probably related to immunity to ticks or resistance to the immunosuppressive effects of ticks. Immunity to dermatophilosis might involve non-classic responses mediated by CD1 antigen presentation and gammadelta T cells. Progress towards vaccination is further complicated by strain-specific acquired immunity to D. congolensis.

Genetic resistance of Creole goats to cowdriosis in Guadeloupe. Status in 1995.

A genetic predisposition to resistance (R)/susceptibility (S) has been demonstrated for cowdriosis in certain goal lines. In order to identify genetic markers of R/S and to follow their transmission to the offspring, 4 groups of sires and dams were crossbred in 1991, 1992, and 1993: Rr, RS, SR, and SS. The offspring were challenged at the age of six months with subsequent challenge of the S parent. From 28 presumed S dams, 7 turned out R and from 2 presumed S sires, 1 was R and the other undefinable. The resistance rate was 76% for 21 RR kids and 45% for 20 RS kids. The R character seems to be better transmitted by dams than the S one. No difference was observed between R and S goats in terms of incubation period and body temperature level after challenge. There was no statistical effect found of sex, litter size, or birth mass on the R/S character; only a slight effect on daily mass gain was observed. Sperm has been collected from 4 S sires (died after challenge) and 5 R sires, and stored in liquid nitrogen in order to conceive additional SR and SS progeny. Several caprine leucocyte antigens seem to be genetic markers of R (Be 1, CLY) and of S (Be 9, Be 22, Be 23). Further powerful tools like microsatellites and probes defining other interesting loci possibly involved in disease pathogenesis are now at hand and will be included in future experiments.

An amino acid sequence coded by the exon 2 of the BoLA DRB3 gene associated with a BoLA class I specificity constitutes a likely genetic marker of resistance to dermatophilosis in Brahman zebu cattle of Martinique (FWI).

One hundred and twenty-seven Brahman cattle from several locations in Martinique (FWI), reared under different environmental conditions, were followed over three years and checked for clinical signs of dermatophilosis. To confirm that these animals had been in contact with the pathogen Dermatophilus congolensis, their sera were tested by ELISA. On the basis of this epidemiological study, 12 animals were classified as resistant (seropositive without clinical signs), belonging to herds in which the prevalence of the disease ranged from 25 to nearly 98%. Eighteen animals classified as highly susceptible displayed severe characteristic skin lesions. These 30 selected animals were typed for class I antigens of the major histocompatibility complex (MHC). MHC class II genes were analyzed using the polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) techniques, on the exon 2 of the bovine leucocyte antigen (BoLA) DRB3 gene. Several alleles were found, according to patterns provided by the restriction enzymes used: Fnu 4HI, Dpn II, Hae III, and Rsa I. A particular sequence "EIAY" at amino acid positions 66/67/74/78 located in the antigen recognition sites (ARS) was found in the 12 animals classified as resistant, and 10 of them displayed also class I BoLA-A8 specificity. On the other hand, only 3 out of the 18 susceptible animals showed simultaneously the BoLA-DRB3 "EIAY" sequence and BoLA-A8 specificity. Interestingly, a serine residue at position 30 of the ARS was found in 8 of the susceptible animals and was completely absent from all resistant animals. Furthermore, in a same animal, the serine at position 30 and the EIAY sequence were never found simultaneously on the same haplotype. These results show a strong correlation between the resistant character to dermatophilosis and the association of MHC haplotypes: the BoLA-A8 specificity and the BoLA-DRB3 "EIAY" sequence at ARS positions 66/67/74/78 with the lack of serine in position 30. To confirm these results, family segregation studies are in progress and some interesting observations have been obtained.

[Modern imaging in cerebral vein angioma]. / L'imagerie moderne dans les angiomes veineux cérébraux.

Cerebral venous angiomas are vascular malformations of the brain which, before the advent of modern imaging, and in particular MRI, were thought to be rare. They have a specific angiographic aspect called "caput Meduse" as it corresponds to the description of a large draining vein to which converge numerous radial veins located in the white matter. These true vascular malformations have characteristic histological features making it possible to differentiate them from other vascular malformations, notably cavernomas. These lesions are less scanty than classically believed, being the most frequent malformations encountered in post-mortem examination series. The circumstances in which cerebral venous angiomas are discovered vary considerably, and haemorrhage can be the revealing sign. Some authors regard these malformations as "abnormalities" of white matter veins and not as pathological entities, since they derive from a change in development during embryogenesis, but this theory is not supported by any anatomical evidence. The same authors attribute the bleeding to an associated cavernoma. Whereas CT enables venous angiomas and possible associated cavernomas to be detected, only repeated MRI explorations performed after the finding of venous angioma and hemorrhage can permit, by analysis of signal changes, to confirm or invalidate the non-pathological theory of the venous malformation and its tendency to bleed. Apart from hemorrhages, since gadolinium is used MRI is the most reliable and least invasive means to diagnose angiomas and to detect associated occult angiographic malformations. For this reason, nowadays only hemorrhages require an angiography in search of an arteriovenous malformation demanding prompt treatment. Moreover, MRI makes it possible to detect angiographically occult malformations.

Protection of goats against heartwater acquired by immunisation with inactivated elementary bodies of Cowdria ruminantium.

In two experiments, four and five goats were vaccinated by giving two subcutaneous injections of a preparation of inactivated elementary bodies of Cowdria ruminantium (Gardel stock) mixed with Freund's adjuvant. All vaccinated animals together with four naive controls were challenged intravenously with 5 ml of supernatant of a culture of bovine endothelial cells infected with the same stock of Cowdria. All goats developed a high temperature. Two out of four, and four out of five vaccinated goats survived the challenge whereas all naive control animals died within 7-12 days. Vaccinated goats which died survived longer than the controls. No difference in antibody titres was observed between protected and non-protected vaccinated goats. Moreover, immune sera from surviving goats, whether heat inactivated or not, were unable to neutralize the infection of bovine endothelial cells by Cowdria in vitro. Mechanisms conferring protection on the immunized goats are unknown at the moment but the hypothesis that T-helper lymphocyte populations have been elicited seems to be likely. This method of immunization with dead organisms will help in the search for protective antigens against cowdriosis.

[Imaging techniques in the diagnosis of brain lesions in eclampsia. Apropos of a case]. / Apport de l'imagerie dans le diagnostic des lésions encéphaliques de l'éclampsie. A propos d'une observation.

Eclampsia is defined as the development of convulsion and/or coma during pregnancy. Physiopathology remains unclear. We report a case of eclampsia and review the literature. CT, MR and angiographic findings are described. Value of various techniques is discussed.