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Purification and characterization of glycosylated and non-glycosylated forms of prolactin from the dromedary (Camelus dromedarius).

1. Camel prolactin (camPRL) was isolated from the insoluble residue left after extraction of the gonadotropins FSH and LH from a single batch of one-humped camels (Camelus dromedarius) pituitaries. 2. Several isoforms of camPRL were isolated and characterized. 3. Glycosylated and non-glycosylated forms of camPRL were separated by affinity chromatography on Con-A Sepharose. 4. The glycosylated form represented more than 20% of total camPRL and was found to be only 10-20% as active as the non-glycosylated form in its ability to bind to rabbit mammary gland PRL receptors and to stimulate the proliferation of Nb2 cells.

Amino Acid Sequence↗

Comparative studies on the distribution of rhodanese in different tissues of domestic animals.

1. The activity of rhodanese in different tissues of some domestic animals was measured. 2. Rhodanese was present in all tissues studied. 3. The activity of rhodanese in most tissues of sheep was higher than other animals studied. 4. In sheep and cattle the epithelium of rumen, omasum and reticulum were the richest sources of rhodanese. Significant activity of rhodanese was also present in liver and kidney. 5. In camel the liver contained the highest level of rhodanese followed by lung and rumen epithelium. Camel liver contained a third of the activity of sheep liver. 6. Equine liver had a third of the activity of sheep liver. Other tissues showed low levels of rhodanese activity. 7. Dog liver contained only 4% of the activity of sheep liver. In this animal, brain was the richest source of rhodanese. 8. The results are discussed in terms of efficacy of different tissues of animals in cyanide detoxification.

Animals↗

An attempt to define the host range for African horse sickness virus (Orbivirus, Reoviridae) in east Africa, by a serological survey in some Equidae, Camelidae, Loxodontidae and Carnivore.

A survey was carried out in horse, zebra, elephant, camel, sheep and goat and wild carnivore sera for virus-serum neutralising antibody to the nine type strains of African horse sickness virus. Antibody was found amongst the horse, zebra and elephant sera to all nine different strains. No antibody was detected in any sera from camels, sheep and goats. None was found in sera from hyaena and jackals in this series but had been detected earlier.

African Horse Sickness↗

Characterization of camelpoxvirus isolates from Africa and Asia.

Five orthopoxvirus isolates of camels from different geographic regions of Africa and Asia were analysed with respect to their biological and genomic attributes. The behaviour of the isolates in various cell cultures, the type of pock lesions on the chorioallantoic membrane of embryonated chicken eggs, and the respective ceiling temperatures were determined. Additionally, physical maps for restriction endonucleases HindIII and XhoI were established. The data obtained from biological assays and DNA analyses demonstrated minor differences between the five isolates. However, these findings confirm previous reports suggesting that orthopoxviruses of camels constitute a separate species within the genus Orthopoxvirus.

Africa↗

Purification and partial characterization of growth hormone from the dromedary (Camelus dromedarius).

Camel growth hormone (camGH) was isolated from the insoluble residue left after extraction of the gonadotropins FSH and LH from a single batch of one-humped camel (Camelus dromedarius) pituitaries. Only one form of camGH was isolated and characterized; no glycosylated form of camGH could be evidenced. The isoelectric points (pI) of camGH was determined by chromatofocusing. The N-terminal amino-acid sequences of camGH was determined and compared to those of GHs from other species. The availability of this hormone and our better knowledge of its structure will permit to undertake the study of its structure-function relationships and of its physiological functions in this economically important species.

Amino Acid Sequence↗

Infertility in the dromedary bull: a review of causes, relations and implications.

Research into infertility in the dromedary bull, as reported during the last two decades, is reviewed with emphasis on causes and effects. Reproductive activity of such animals is naturally limited by a breeding season, though with enough encouragement some may mate with oestrous females out of season but a full fertilization potential can in no way be expected. It is essential that any female presented to a bull is capable of reproducing. The presentation of a subfertile or infertile female due to infection or physiological abnormality will adversely affect the female's ability to conceive and, therefore, the apparent fertility rates of the bull she was put to. The average number of successful services a bull could be expected to perform is two per day. Dromedary bulls with large testes have larger sperm outputs and can cope with more than two females per day providing that they are given adequate periods of rest, 1-2 days every 10 days or so, in conjunction with appropriate nutrition throughout the season. Anabolic steroids or testosterone therapies, which are sometimes used in an attempt to improve male characteristics and bull libido, are not recommended for dromedary bulls in breeding work. Such steroids result in a decrease in testicular size and weight with fewer sperm per gram of testicular tissue being found and the sperm produced also have lower motility rates. Pain associated with the act of mating a she-camel, due to injuries or inflammation in the scrotum, testes, prepuce and sheath, can cause a permanent reduction in bull libido. Camel bulls achieving pregnancy rates more than 60% have had consistently higher spermatozoal concentrations and kinematic variables derived by the computerized cell motion analyzer (CMA) system. As far as physical capabilities are concerned, 3-year-old dromedary bulls, which have reached puberty, have been shown to be perfectly capable of fertilizing a female, but they have a limited sperm production to perform consistently throughout the season in a large herd. By 4.5-5.0 years of age, they are capable of producing adequate numbers of sperm to mate as many as females as an adult bull but fertilizing capacity is not attained until 6 years of age on average. Hyperoestrogenaemia, associated with autoimmune thyroiditis and trypanosomiasis, suppresses the secretion of testicular testosterone and augments the release of testicular histamine, which appears mandatory for quantitative reduction/loss of advanced spermatogenic cells in infertile dromedary bulls.

Aging↗

Isolation and characterization of raw heparin from dromedary intestine: evaluation of a new source of pharmaceutical heparin.

Heparin, a heterogeneous anionic polysaccharide, is the glycosaminoglycan (GAG) used clinically an anticoagulant. This anticoagulant activity is primarily derived from its binding to the serine protease inhibitor antithrombin III, a potent inhibitor of thrombin (factor IIa) and factor Xa. Heparin is a complex natural product and its in vitro synthesis is not yet possible due to the difficulty of organizing the many biosynthetic enzymes required for its synthesis. The principle natural sources for heparin include porcine intestine and bovine lung. These two sources pose concerns for religious and health reasons, respectively. To circumvent these concerns, GAG from the intestinal tissue of one humped camel was isolated. Chemical characterization of this newly isolated GAG and spectroscopic analysis by 1D and 2D 1H-NMR were undertaken. Unsaturated disaccharide compositional analysis was performed on the enzymatically depolymerized GAG and the molecular weight of the isolated GAG was determined by gradient polyacrylamide gel electrophoresis. Anticoagulant activity of the newly isolated GAG was tested by using an anti-factor Xa assay. The results of these studies suggest that the GAG from one humped camel intestine is a mixture of heparin and heparan sulfate and represents an alternative source of heparin.

Animals↗

A PCR system for detection of species and genotypes of the Echinococcus granulosus-complex, with reference to the epidemiological situation in eastern Africa.

We describe the development of a specific and sensitive PCR/semi-nested PCR system for the rapid diagnosis of Echinococcus granulosus genotype G1, E. granulosus genotype G6/7, and Echinococcus ortleppi (G5). Diagnosis of G1 and the group G5/6/7 is performed by a simple PCR, while discrimination between E. ortleppi (G5) and G6/7 involves a subsequent semi-nested PCR step. The target sequence for amplification is part of the mitochondrial 12S rRNA gene. Specificity of the PCRs was 100% when evaluated with isolates of 16 species of cestodes, including Echinococcus multilocularis, Echinococcus equinus, E. ortleppi and three strains of E. granulosus (G1, G6 and G7). Sensitivity threshold was 0.25pg of DNA. This new approach was compared with published protocols of restriction fragment length polymorphism-PCR and sequencing of mitochondrial cytochrome c oxidase subunit 1 and NADH dehydrogenase 1 genes using Echinococcus isolates of human, sheep, goat, camel, cattle and pig origin from Kenya and Sudan. Additionally, two internal DNA probes were developed, one hybridising only with G1, the other with G5, G6 and G7 amplification products. Preliminary epidemiological results obtained with this PCR approach include the detection of a camel strain (G6) infection for the first time in a human patient from eastern Africa, and the first reports of E. ortleppi (G5) in livestock from Kenya and the Sudan.

Animals↗

Cattle-rangeland management practices and perceptions of pastoralists towards rangeland degradation in the Borana zone of southern Ethiopia.

A survey was conducted in the Borana pastoral areas of southern Ethiopia to assess current livestock production systems, rangeland management practices and the perceptions of the pastoralists towards rangeland degradation. This information is considered vital to future pastoral development planning and interventions. Data were collected from a total of 20 villages that were identified from 5 peasant associations, namely Did Yabello, Moyatte, Did Harra, Dubuluk and Melbana. The average household size in the study area was 7.23. The majority of the pastoralists relied on both livestock and crop farming. The average livestock holding per household was 14 cattle, 10 goats, 6 sheep and 2 camels. Livestock holdings, with the exception of camels, has shown a declining trend over time. The two most important traditional rangeland management strategies adopted by the pastoralists included burning and mobility, but since 1974/75 burning has no longer been practised. With regard to mobility, the livestock herding falls in two categories, namely: home based and satellite herding. The former involves the herding of milking cows, calves and immature animals (<2 years) close to the encampments. The latter constitutes the herding of bulls and immatures (>2 years) further away from the encampments. Based on the pastoralists' perceptions, the major constraints on livestock production in descending order, were recurrent drought, feed and water scarcity, animal diseases, predators and communal land ownership. All the respondents considered the condition of the rangelands to have declined dramatically over time. In the past most development policies were based on equilibrium theories that opposed the communal use of the rangelands and traditional range management practices. The way in which the pastoral system affects the rangeland ecosystem functioning is contentious to this theory and the 'tragedy of the commons'. There was also a perceived problem of bush encroachment and the ban on traditional burning practices and recurrent droughts were seen as aggravating factors to this invasion process. The increasing practice of crop cultivation on the rangelands was identified as a serious threat to future livestock production and traditional resource management practices. Suggestions for possible interventions to improve the productivity and sustainable use of these rangelands are made.

Adult↗

Characterization of Trypanosoma evansi type B.

A distinctive feature of Trypanosoma evansi is the possession of a kinetoplast that contains homogeneous DNA minicircles, but lacks DNA maxicircles. Two major sequence variants of the minicircle have been described and here we have sequenced the type B variant and designed a specific PCR test to distinguish it from type A. Further a test based on maxicircles to distinguish T. brucei brucei from T. evansi was designed and evaluated. Using the designed PCR tests, we detected three type B isolates from camel blood samples collected in northern Kenya, more than 20 years after the first isolation of type B. Comparison of minicircle sequences from all four type B isolates shows >96% identity within the group, and 50-60% identity to type A minicircles. Phylogenetic analysis based on minicircle sequences reveals two clusters, one comprising isolates of type A and one of type B, while random amplification of polymorphic DNA show slight polymorphic bands within type B. Most T. evansi isolates analysed were heterozygous at a repetitive coding locus (MORF2). All type B isolates had one genotype designated 3/5 based on the alleles present. Three camel isolates, which had homogenous type A minicircles, lacked the RoTat 1.2 gene, while another five isolates were T. b. brucei, based on the heterogeneity of their minicircles and presence of maxicircles as demonstrated by PCR amplification of the gene for cytochrome oxidase subunit 1. Our results confirm the existence of T. evansi type B isolates, T. b. brucei and existence of T. evansi type A without RoTat 1.2 gene in Kenyan isolates.

Animals↗

Epidemiology and control of echinococcosis in sub-Saharan Africa.

Cystic echinococcosis (CE) is highly endemic among the nomadic pastoral tribes of East Africa, but is rare amongst the agriculturally based communities. Echinococcus granulosus infections are common in dogs from all countries in sub-Saharan Africa where they have been examined. Transmission of E. granulosus to humans is affected by such factors as prevalence of the parasite in domestic dogs, behaviors of humans towards dogs, and heterogenicity of the parasite and susceptibility of humans to infection. Sheep and goats appear to be the most common domestic intermediate hosts, but recent studies suggest that camels are equally important intermediate host, especially in Sudan and Turkana. At least five of ten E. granulosus genotypes are infective to humans in sub-Saharan African. Most human cases of CE are caused by the sheep strain (GI) and camel strain (G6) of E. granulosus. Other strains occurring in the area may include a lion strain, the horse strain (G4 or Echinococcus equinus) and the cattle strain (G5 or Echinococcus ortleppi).

Africa South of the Sahara↗

Beta-endorphin: demonstration of binding sites in three human neuroblastoma cell lines specific for the COOH-terminal segment of the human hormone.

This communication reports for the first time specific binding sites in human neuroblastoma cells for human beta-endorphin (beta h-EP). Three cell lines (IMR-32, NMB and Kelly) were investigated and found to bind tritiated beta h-EP with an apparent dissociation constant of 2.2-4.2 nM. Further characterization with camel beta-EP and synthetic analogs indicated that the binding is most likely mediated by the COOH-terminal segments. beta h-EP-(6-31) had significant potency (15-75%) and beta h-EP-(1-27) was without displacing activity. The camel beta-EP has below 1% of the human beta-EP activity.

Binding, Competitive↗

ITS-1 ribosomal DNA sequence variants are maintained in different species and strains of Echinococcus.

This study investigated sequence heterogeneity in the first internal transcribed spacer (ITS-1) of ribosomal DNA within and among species and strains of Echinococcus. Different ITS-1 sequence variants exist in Echinococcus granulosus and Echinococcus multilocularis, which represent at least four evolutionary lineages: (1) a sheep strain-lineage of E. granulosus, (2) a sister lineage of a cervid and camel E. granulosus ITS-1 variants, (3) a lineage including the ITS-1 variants representing horse, bovine and camel strains of E. granulosus, as well as variants from E. multilocularis, Echinococcus oligarthrus and Echinococcus vogeli and (4) a distinct lineage of ITS-1 variants including E. granulosus strains from sheep and cervid, and E. multilocularis. At least two of the species (E. granulosus and E. multilocularis) were paraphyletic for ITS-1. Divergent ITS-1 variants from these two species shared distinct evolutionary lineages. The sequence data provided evidence that at least two turnover mechanisms, namely slippage and unequal crossing over/transposition, have led to the divergence and maintenance of sequence variants in Echinococcus species and strains.

Animals↗

Induction of immune responses and molecular cloning of the heavy chain antibody repertoire of Lama glama.

Functional heavy chain immunoglobulins have, so far, only been found in camels and llamas. Antigen-specific fragments of these heavy chain IgGs (V(HH)) are of great interest in biotechnology because they are very stable and can be produced at high level by the yeast Saccharomyces cerevisiae. The work described in this paper was conducted to determine whether llamas (Lama glama) are a practical source of antigen-specific V(HH) fragments. Llamas were immunised with various types of antigens and the antibody responses were examined during the course of immunisation. Both, conventional and heavy chain IgG antibodies were produced in response to each of the antigens. The heavy chain IgG repertoire displayed a recognition pattern different to that of conventional llama IgGs, resulting in the expansion of the accessible epitope repertoire. Llamas have a lower proportion of heavy chain IgG antibodies in their serum than have camels. To enable the specific and efficient isolation of V(HH) genes from peripheral blood B-cells, the long and short-hinge sequences of Lama glama heavy chain IgGs were determined, revealing the presence of a novel subclass of short-hinge heavy chain IgG. Long and short-hinge specific PCR primers were designed to be used in the construction of llama V(HH) libraries. We conclude that, using the techniques described, antigen-specific V(HH) antibody fragments are readily accessible from the llama, thus providing highly valuable binding molecules for a variety of applications.

Amino Acid Sequence↗

Echinococcus granulosus antigen B and seroreactivity in natural ovine hydatidosis.

Hydatid cyst fluid from sheep and camels infected with Echinococcus granulosus, together with partially purified preparations of hydatid fluid antigen B and a recombinant antigen B product, were tested in an ELISA for their ability to detect IgG antibodies against E granulosus in the serum of naturally infected sheep. The antibody activity in sera from sheep naturally infected with Taenia hydatigena cysticercosis or Fasciola hepatica was also tested. All the antigen preparations from native hydatid cyst fluid were able to detect antibodies in the sera from a significant proportion of sheep with natural hydatid cyst infection, as identified by inspection at slaughter, although the seroreactivity was variable. The native antigen B preparation from camel hydatid cyst fluid gave the highest sensitivity in the ELISA (total 90 per cent), with 99 per cent specificity. In all cases, the recombinant antigen B was the least sensitive antigen (25 per cent) although it was highly specific (99 per cent).

Animals↗

Detection of antibodies to Leishmania donovani in animals in a kala-azar endemic region in eastern Sudan: a preliminary report.

The prevalence of antibodies against Leishmania donovani in selected domestic and wild animal species in 2 villages in Sudan with active L. donovani transmission in humans was investigated. Screening of domestic animals (donkeys, cows, sheep, goats, camels and dogs) with the direct agglutination test (DAT) detected reaction rates above the cut-off titres in donkeys (68.7%), cows (21.4%) and goats (8.5%), and which were also found in wild rats (5.5%). Sera of sheep, camels and dogs had a weak agglutination reaction below the cut-off titre. Testing of the same sera by enzyme-linked immunosorbent assay (ELISA), against a lysate of L. donovani promastigotes, showed reaction rates above the cut-off optical density in cows (47.6%), goats (13.6%), and in rats (4.1%). No Leishmania parasite was isolated from spleen, liver, bone-marrow or spleen of Nile rats.

Animals↗

Crimean-Congo haemorrhagic fever virus infection in the western province of Saudi Arabia.

In 1990, an outbreak of suspected viral haemorrhagic fever involving 7 individuals occurred in Mecca in the Western Province of Saudi Arabia. Congo-Crimean haemorrhagic fever (CCHF), not previously known to be present in Saudi Arabia, was incriminated. A study of the epidemiology of this virus was therefore carried out in Mecca, and in nearby Jeddah and Taif in 1991-1993; 13 species of ixodid ticks (5 Hyalomma spp., 5 Rhipicephalus spp., 2 Amblyomma spp., 1 Boophilus sp.) were collected from livestock (camels, cattle, sheep, goats), and of these 10 were capable of transmitting CCHF. Camels had the highest rate of tick infestation (97%), and H. dromedarii was the commonest tick (70%). Attempts to isolate virus from pools of H. dromedarii and H. anatolicum anatolicum were unsuccessful. The source of infection in 3 confirmed cases of CCHF was contact with fresh mutton and, in a suspected case, slaughtering sheep. An investigation in Mecca, which included a serological survey of abattoir workers, identified 40 human cases of confirmed or suspected CCHF between 1989 and 1990, with 12 fatalities. Significant risk factors included exposure to animal blood or tissue in abattoirs, but not tick bites. It is suspected that the CCHF virus may have been introduced to Saudi Arabia by infected ticks on imported sheep arriving at Jeddah seaport, and that it is now endemic in the Western Province.

Adult↗

Emergence and evolution of functional heavy-chain antibodies in Camelidae.

Antibodies of jawed-vertebrates are composed of paired heavy (H) and light (L) polypeptide chains. Surprisingly, the sera of camelids, nurse shark and wobbegong shark, and possibly ratfish contain antibodies that lack L-chains. In camelids, these Heavy-chain antibodies (HCAbs) are gamma-isotypes, and are functional in antigen binding. In this review we focus on the dedicated immunoglobulin (Ig) genes that encode the HCAb in Camelidae (camels, dromedaries and llamas), about their origin, and how these camel immunoglobulins evolved and acquire a large and diverse repertoire of antigen binding sites in absence of the H-L combinatorial diversity.

Amino Acid Sequence↗