Bovine SLC11A1 3' UTR SSCP genotype evaluated by a macrophage in vitro killing assay employing a Brucella abortus strain

The 3' untranslated region (3' UTR) of the bovine natural resistance-associated macrophage gene (NRAMP1 or SLC11A1) was genotyped in Colombian Creole Blanco Orejinegro (BON) (Bos taurus) (n = 140) and Zebu Brahman (Bos indicus) (Z) (n = 20) cattle and their crosses (BON x Zebu Brahman [B x Z] [n = 10]; Zebu Brahman x BON [Z x B] [n = 10]), and in animals from a Holstein x BON (H x B) (n = 10) cross. Direct sequencing and single-strand conformation polymorphism analysis (SSCP) helped in detecting the polymorphic behaviour. The association between resistance to brucellosis infection and SSCP genotype was evaluated using a macrophage in vitro killing assay employing a virulent Brucella abortus strain. The 3' UTR (GT) repeated polymorphism was gentoyped and its association with resistance to brucellosis was evaluated. When all breeds were grouped, a high frequency in the homozygote GT(12) (AA genotype) (0.823) and a very low frequency in the homozygote GT(10) (BB genotype) (0.047) were detected. The BON (0.963), Z x B (0.60) and H x B (1.00) cattle showed high GT(12) allele frequencies, unlike that seen for the B x Z and Zebu cattle (0.3002 and 0.218, respectively). The GT(10) allele was only found in the Zebu cattle (0.391). A significant association (p < 0.001) was found between the B. abortus macrophage in vitro killing assay phenotypes and the bovine SLC11A1 3' UTR genotypes, which suggests that the A allele may be associated with resistance. Because only nine animals had the BB genotype, the results require some confirmation in more extensive populations.     

Novel variants within the coding regions of the Slc11A1 gene identified in Bos taurus and Bos indicus breeds

Although in the cow the genetic resistance to brucellosis has been previously attributed to the Slc11A1 gene encoding Nramp1 protein, none of the mutations described to date seems to be the cause. To be able to associate another polymorphism of the gene to brucellosis resistance, we characterized the gene and identified in different breeds of Bos taurus and Bos indicus, six new variants among a total of 11 single nucleotide mutations, of which five occurred in the coding sequence (three are missense mutations), one in the promoter region and five in introns. The allelic and genotypic frequencies calculated revealed differences (p < 0.05) among the breeds studied.     

Pedigree analysis in the Brazilian Zebu breeds

Zebu breeds play an important role in cattle production systems in Brazil. To assess the genetic variability from animals in the Herd Books of Nelore, Gir and Guzerat breeds, generation intervals, inbreeding, effective population size and parameters of gene origin (effective number of founders, ancestors and founder genomes) were calculated using pedigree records from 1938 to 1998. Breed subdivision was quantified by Wright's F -statistics. Calculations were separately carried out for consecutive 4-year intervals in the period 1979–98. Generation interval was around 8 years for the three breeds. Total inbreeding increased in all the breeds reaching values of 2.13%, 2.28% and 1.75%. Effective population size decreased from 85 to 68 in Nelore, from 70 to 45 in Gir and remained nearly constant around 104 in Guzerat. The quantities assessing the number of contributing ancestors decreased with time in all the breeds, and in the last analysed period the most important ancestor accounted for 14%, 3.1% and 4.1% in Nelore, Gir and Guzerat, respectively. Results indicate that the studied breeds are suffering from a loss of genetic variability which can result in negative effects on breeding and conservation purposes.     

Promoter region of the bovine growth hormone receptor gene: single nucleotide polymorphism discovery in cattle and association with performance in Brangus bulls

Expression of the GH receptor (GHR) gene and its binding with GH is essential for growth and fat metabolism. A GT microsatellite exists in the promoter of bovine GHR segregating short (11 bp) and long (16 to 20 bp) allele sequences. To detect SNP and complete an association study of genotype to phenotype, we resequenced a 1,195-bp fragment of DNA including the GT microsatellite and exon 1A. Resequencing was completed in 48 familialy unrelated Holstein, Jersey, Brown Swiss, Simmental, Angus, Brahman, and Brangus cattle. Nine SNP were identified. Phylogeny analyses revealed minor distance (i.e., <5%) in DNA sequence among the 5 Bos taurus breeds; however, sequence from Brahman cattle averaged 27.4 +/- 0.07% divergence from the Bos taurus breeds, whereas divergence of Brangus was intermediate. An association study of genotype to phenotype was completed with data from growing Brangus bulls (n = 553 from 96 sires) and data from 4 of the SNP flanking the GT microsatellite. These SNP were found to be in Hardy-Weinberg equilibrium and in phase based on linkage disequilibrium analyses (r(2) = 0.84 and D'= 0.92). An A/G tag SNP was identified (ss86273136) and was located in exon 1A, which began 88 bp downstream from the GT microsatellite. Minor allele frequency of the tag SNP was greater than 10%, and Mendelian segregation was verified in 3 generation pedigrees. The A allele was derived from Brahman, and the G allele was derived from Angus. This tag SNP genotype was a significant effect in analyses of rib fat data collected with ultrasound when bulls were ~365 d of age. Specifically, bulls of the GG genotype had 6.1% more (P = 0.0204) rib fat than bulls of the AA and AG genotypes, respectively. Tag SNP (ss86273136), located in the promoter of GHR, appears to be associated with a measure of corporal fat in Bos taurus x Bos indicus composite cattle.     

Characterization of breeds of Bos indicus and Bos taurus cattle for maternal and individual traits

Angus, Boran and Red Poll sires were mated to Ankole, Boran and small East African Zebu (Zebu) females to produce the cows characterized in this study, which produced calves by Friesian, Brown Swiss, Simmental, Boran and Red Poll sires. Individual traits included cow parturition weight, cow weight at weaning, cow mean weight and cow weight change from parturition to weaning; maternal traits included calf crop born percentage, preweaning viability percentage, overall viability percentage, birth weight, weaning weight, weight at 12, 18 and 24 mo and calf weight weaned per cow exposed to breeding (productivity index). Cows by Angus and Red Poll sires significantly exceeded cows by Boran sires (all cows in this comparison had Ankole and Zebu dams) in weaning weight of progeny and in cow weight at parturition and at weaning. Cows by Red Poll sires significantly exceeded cows by Boran sires in birth weight of progeny. Progeny of cows by Boran sires significantly exceeded progeny of cows by Angus and Red Poll sires in 24-mo weight. Cows by Angus and Red Poll sires did not differ (P greater than .05) in any of the traits analyzed. Cows with Ankole and Boran dams significantly exceeded cows with Zebu dams (all cows in this comparison had Angus and Red Poll sires) in progeny weight at birth, 18 mo and 24 mo and in cow weight at parturition and at weaning. Cows with Boran dams significantly exceeded cows with Zebu dams in weaning and 12-mo weight of progeny and significantly exceeded cows with Ankole dams in weaning weight of progeny. The significantly heavier weaning weight and significantly lighter 24-mo weight of the 3/4 Bos taurus progeny of cows with Angus and Red Poll sires relative to the 1/2 Bos taurus progeny of cows with Boran sires suggest that cattle that are 1/2 Bos taurus breed composition have greater general adaptation than cattle with 3/4 Bos taurus breed composition in the postweaning nutritive and climatic environment under which this experiment was conducted. Significantly heavier weights of Angus and Red Poll crossbred cows relative to Boran crossbred cows (all breeds crossed on Ankole and Zebu cows) indicate that the two Bos taurus breeds exceed the Boran (Bos indicus) breed in additive direct genetic effects for size when they have general adaptation to the environment.     

Sequence analysis of the NRAMP1 genes from different bovine and buffalo breeds

The natural resistance associated macrophage protein 1 (Nramp1) has been reported to confer resistance or susceptibility to Mycobacterium bovis, Salmonella typhimurium, and Leishmania donovani in the mouse, Mus musculus. A Gly and Asp substitution at position 169 of the mouse Nramp protein is invariably associated with the resistant and susceptible phenotypes, respectively. The present study aimed to detect polymorphisms in the NRAMP1 gene from different cattle and buffalo breeds. Genomic DNAs from five breeds of cattle and four breeds of buffalo were used in the study. Sequencing showed two nucleotide substitutions found in intron 4, three in exon V, and ten in intron 5. An amino acid substitution was observed at nucleotide position 1202 in exon V of the Japanese black, Angus, Philippine and Bangladesh swamp-type buffaloes which coded for Thr, while the Korean cattle, Holstein, African N'dama, Indonesian swamp-type buffalo and the Bangladesh river-type buffalo had Ile. All the breeds of cattle and buffaloes tested in this study coded for Gly at the position in exon VI which corresponds to the same amino acid of the murine Nramp1-resistant phenotype at position 169. The phylogenetic relationship among the different breeds showed a cluster comprised mainly of cattle and another one mainly of buffaloes.     

红安格斯牛mtDNA D-loop序列变异

测定8头红安格斯牛mtDNA D-loop全序列911bp,确定的6种单倍型含有49个变异位点,1次颠换,46次转换,明显的转换倾向,2次缺失/插入,其中5种单倍型为普通牛线粒体单倍型,属普通牛支系,而Angus-6属瘤牛支系,表明红安格斯牛兼含有普通牛和瘤牛的遗传背景,但受瘤牛的影响非常小,和大多现代普通牛一样,红安格斯牛受到过瘤牛基因渐渗的影响.     

Some preliminary observations on the susceptibility and resistance of different cattle breeds to Theileria parva infection

Theileria parva-na?ve Friesian (Bos taurus), Boran (Bos indicus) and Maasai Zebu steers (B. indicus) were infected with a T. parva sporozoite stabilate dose which had previously been shown to induce an estimated 50% mortality rate in Boran cattle. All the cattle developed patent infections with no significant differences in the length of the prepatent period to development of macroschizonts (P > 0.05) between the three groups. Clinical theileriosis occurred in all eight the Friesians (100%), five out of nine Borans (55.6%) and two out of five Zebus (40%). Three of the Friesians (37.5%), and two of the Borans (22.2%) died of theileriosis. The different cattle types were equally susceptible to the infective dose used as indicated by the length of the prepatent periods, but there was a marked difference in their development of clinical theileriosis. The gradation in resistance to disease confirms the findings of earlier less critical studies and identifies these cattle breeds as suitable for investigations into the mechanisms of resistance to theileriosis.     

Susceptibility and immune responses of zebu and taurine cattle of West Africa to infection with Trypanosoma congolense transmitted by Glossina morsitans centralis

Following tsetse-transmitted infection with Trypanosoma congolense, major differences in development of localised skin reactions, the ability to control parasitaemia, the degree of anaemia and in antibody response to trypanosomes were found between the reputedly trypanotolerant breeds of cattle (N'Dama, N'Dama/Baoule crosses, Baoule) and the trypanosusceptible West African Zebu. The local skin reactions that developed in the Zebu were large and severe while those that occurred in the other breeds were smaller and less severe or mild. The timing of appearance of parasitaemia and the height of the first peaks were similar in all the animals, but the Zebu were less able to control subsequent waves of parasitaemia. Possibly reflecting these events, it was only in the Zebu that significant anaemia developed. Neutralizing antibody against homologous metacyclic trypanosomes developed between 14 to 18 days after infection in all breeds of cattle; however, marked differences were found when antibody to trypanosomes derived from first peak parasitaemias were tested in the Zebu and Baoule. Neutralizing antibody against these parasites appeared in the Baoule on day 24 but were not detected in Zebu until day 51. Furthermore, the antibody titres were 3 log2 higher in the Baoule. It was concluded that the trypanotolerance exhibited by the West African taurine cattle might be related to a) their ability to control trypanosome numbers in the skin and in the bloodstream, an outcome that was possibly brought about by the earlier and superior immune response and b) failure to develop anaemia which might be associated with their capacity to control parasitaemia.     

中国黄牛Y-STRs遗传多样性与起源研究

[目的]研究中国黄牛Y染色体STRs的遗传多样性及父系起源。[方法]利用非变性聚丙烯酰胺凝胶电泳,选择2个牛Y-STRs位点INRA189和BM861,分析16个中国地方黄牛品种284头公牛与4头缅甸黄牛公牛的Y染色体遗传多样性。[结果]在中国16个黄牛品种中,2个Y-STR位点可以区分中国黄牛中的普通牛和瘤牛类型,表明中国黄牛有普通牛和瘤牛两种父系起源。4头缅甸黄牛均为瘤牛类型。在中国16个黄牛品种中,普通牛和瘤牛分布频率分别为57.0%和43.0%,其中普通牛频率在北方黄牛中占优势(98.3%),瘤牛频率在南方黄牛中占优势(76.1%),中原黄牛中普通牛频率较高为63.8%,瘤牛频率为36.2%。[结论]中国黄牛存在普通牛和瘤牛两种父系起源;普通牛频率自北向南逐渐减少,瘤牛频率自北向南逐渐增加,中原地区为普通牛和瘤牛的交汇处。     

The blood volumes and erythrokinetics of Ndama and Zebu cattle experimentally infected with Trypanosoma brucei.

The responses of susceptible Ndama and Zebu cattle to experimental infection with Trypanosoma brucei were compared using haematological, parasitological and radioisotopic methods. Animals of both breeds became anaemic, but this was more severe in the Zebu cattle, one of which died. Although the prepatent period was the same in animals of both breeds, the levels of the first and subsequent peaks of parasitaemia were higher in the Zebu. The anaemia was due to an accelerated rate of red cell break-down which was more marked in the Zebu cattle. Haemodilution was not a feature. There was no evidence of dyshaemopoiesis but iron reutilisation from degraded erythrocytes was impaired. The greater resistance of the Ndama to T brucei infection could not be attributed to the capacity of this breed to mount a more effective erythropoietic response than the Zebu.     

Comparative Thickness of the Medulla of Bos taurus and Bos indicus Kidneys

Twenty kidneys of 4 years old Bos taurus cattle and 26 kidneys of 2—4 years old Bos indicus (Zebu cattle) of East African Shorthorn breen were examined lobe by lobe after fixation. The cortex and medulla with subunits were measured and compared in the two sub-species. The width of the cortex was about the same in both. The outer and inner stripes of the outer medullary zone were significantly broader in Bos taurus whereas outer inner zone was significantly broader in Bos indicus. The width of the entire medulla was greater in Bos taurus.     

Identification and characterization of polymorphisms within the 5' flanking region, first exon and part of first intron of bovine GH gene

The aim of the present study was to identify and characterize polymorphisms within the 5′ flanking region, first exon and part of first intron of the bovine growth hormone gene among different beef cattle breeds: Nelore (n = 25), Simmental (n = 39), Simbrasil (n = 24), Simmental × Nelore (n = 30), Canchim × Nelore (n = 30) and Angus × Nelore (n = 30). Two DNA fragments (GH1, 464 bp and GH2, 453 bp) were amplified by polymerase chain reaction and then used for polymorphism identification by SSCP. Within the GH1 fragment, five polymorphisms were identified, corresponding to three different alleles: GH1.1, GH1.2 and GH1.3 (GenBank: AY662648 , AY662649 and AY662650 , respectively). These allele sequences were aligned and compared with bovine GH gene nucleotide sequence (GenBank: M57764 and AF118837 ), resulting in the identification of five insertion/deletions (INDELs) and five single nucleotide polymorphisms (SNPs). In the GH2 fragment two alleles were identified, GH2.1 and GH2.2 (GenBank: AY662651 and AY662652 , respectively). The allele sequences were compared with GenBank sequences ( M57764 , AF007750 and AH009106 ) and three INDELs and four SNPs were identified. In conclusion, we were able to identify six new polymorphisms of the bovine GH gene (one INDEL and five SNPs), which can be used as molecular markers in genetic studies.     

Parasite burdens and host susceptibility of Zebu and N'Dama cattle in village herds in Gambia

Helminth and tick burdens of N'Dama cattle kept in village herds were compared with parasite burdens in Zebu cattle introduced into the same herds. The animals were monitored regularly for tick and helminth burdens, and blood samples were examined for differential white cell counts. The Zebu were found to have significantly higher numbers of endo- and ectoparasites. The N'Dama had significantly higher levels of circulating eosinophils than the Zebu during peak parasite burdens.

No difference in antibody levels to Amblyomma variegatum infestation could be detected between the two breeds. It is suggested that observed differences in susceptibility to endo- and ectoparasites between the two breeds cannot be totally attributed to environmental naivety of the Zebu, but must be partially owing to enhanced innate resistance factors in the N'Dama.     

Genetic diversity of growth hormone receptor gene in cattle

Growth hormone receptor (GHR) belongs to a member of the cytokine receptor superfamily. Polymorphism of presence or absence of an approximately 1.2 kbp LINE-1 element is observed in bovine GHR gene. The present study was carried out for estimating the genetic diversity and the origin of the LINE-1 element in 10 European, Southeastern Asian and East Asian cattle breeds or populations. Genotyping of the LINE-1 revealed predominant LINE-1 presence in European breeds (0.917∼0.991), absence in the Bos taurus indicus populations (0.000∼0.017), and intermediate presence in Northeast Asian cattle (0.417∼0.522). From genetic features of LINE families, LINE-1 of GHR could be attributed to the same origin in both European and Asian cattle, and Asian LINE-1 may not be derived from recent introgression. This result suggested that LINE-1 in bovine GHR gene could have arisen in an ancestral population of Bos taurus taurus .     

Susceptibility of Bos indicus and Bos taurus to Anaplasma marginale and Babesia bigemina infections

The reaction of Bos taurus and pure-bred Bos indicus heifers to infection with the intraerythrocytic parasites Anaplasma marginale and Babesia bigemina was studied. B. bigemina infection at 18 months and A. marginale infection at 13 or 24 months resulted in slightly less severe reactions in pure-bred Bos indicus cattle than in Bos taurus. In both breeds, the reaction to A. marginale infection was more severe in older cattle. The severity of B. bigemina infection was not affected by a previous infection with A. marginale.     

Carrier rate of Factor XI deficiency in stunted Japanese black cattle

Blood examinations and genotyping of Factor XI (F11) were performed in growth retardation Japanese Black cattle and their dams. Genotyping of F11 revealed that the recessive homozygous and heterozygous genotype frequencies were 5.2% and 50.0% in the Claudin-16 (CL-16) deficiency group (n=58), 0% and 14.2% in the renal dysplasia group (n=7), 0% and 26.1% in the non-CL-16 deficiency nephritis group (n=23), 8.9% and 46.7% in the hypogenesis syndrome group (n=45), 6.2% and 25.0% in the neonatal weak calf syndrome group (n=32), 9.1% and 38.6% in the respective dams group (n=44), 0% and 23.1% in the normal cattle group (n=13), and 5.9% and 38.2% in total (n=222), respectively. These results showed that the carrier rate of F11 deficiency was high in Japanese Black cattle, and that the CL-16 deficiency, hypogenesis syndrome, neonatal weak calf syndrome, and dams groups had a large amount of recessive homozygous genotype than the other groups. No abnormal bleeding was observed clinically in the present study, and 4 of the recessive homozygous dams showed normal growth and parturition.     

Chromosomal localisation and genetic variation of the SLC11A1 gene in goats (Capra hircus)

The solute carrier family 11 member A1 (SLC11A1) gene is associated with resistance to infectious diseases. Chromosomal localisation, genomic regions corresponding to functional domains and the genetic variability of microsatellites in the 3' untranslated region (3'-UTR) of this gene were investigated in 427 goats (Capra hircus) of six breeds. Using dual colour fluorescence in situ hybridisation, SLC11A1 was localised to goat chromosome 2. Single strand conformation polymorphism was used to screen for polymorphisms in SLC11A1 exons 2, 10 and 15. There was no variation among goat breeds in the sarcoma homology 3 (SH3) binding motif, the protein kinase C phosphorylation site or the two N-linked glycosylation sites. Exon 15 exhibited variability due to the presence of two polymorphic microsatellites. Genotyping of the upstream guanine-thymine repeat (GTn) at 3'-UTR revealed eight alleles (GT11, GT12, GT14-GT19) in goats, whereas GT13 (present in cattle) was absent. Most goats carried the GT16 allele and no allele was found to be exclusive to only one breed. The coefficient of genetic differentiation value (G(ST)) was 0.084. This microsatellite appears to be an informative DNA marker for genetic linkage analysis in goats.