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detection of mycobacterium avium subspecies paratuberculosis by is900-based pcr assays from an alpaca (vicugna pacos) kept in a german zoological garden.an alpaca (vicugna pacos) bred and kept in a german zoological garden exhibited clinical signs consistent with paratuberculosis. the presence of mycobacterium avium ssp. paratuberculosis (map) was confirmed in feces and in the ileocecal lymph node (iln) by is900-based polymerase chain reaction (pcr) assays and culture. a bacterial burden of 7.6 x 10(6) map/g in feces and 4.4 x 10(7) map/g in lymph node tissue was determined by real-time pcr. for further characterization, a conventional pcr was d ...201323505723
spatial point pattern analyses of bovine viral diarrhea virus infection in domestic livestock herds and concomitant seroprevalence in wild white-tailed deer (odocoileus virginianus) in new york state, usa.bovine viral diarrhea virus (bvdv) is an economically important disease of domestic cattle that is capable of infecting cervids. a first step in the formulation of a regional bvdv management plan is a local assessment of the likelihood of pathogen transmission from wildlife to domestic livestock. to achieve this, blood samples were collected from hunter-harvested white-tailed deer (odocoileus virginianus) throughout new york state in the fall of 2009 and 2010. the svanovir bvdv p80-ab enzyme-lin ...201323512919
anp and bnp responses to dehydration in the one-humped camel and effects of blocking the renin-angiotensin system.the objectives of this study were to investigate and compare the responses of atrial natriuretic peptide (anp) and b-type natriuretic peptide (bnp) in the circulation of hydrated, dehydrated, and dehydrated losartan - treated camels; and to document the cardiac storage form of b-type natriuretic peptide in the camel heart. eighteen male camels were used in the study: control or hydrated camels (n = 6), dehydrated camels (n = 6) and dehydrated losartan-treated camels (n = 6) which were dehydrated ...201323516417
production, crystallization and x-ray diffraction analysis of two nanobodies against the duffy binding-like (dbl) domain dbl6ℇ-fcr3 of the plasmodium falciparum var2csa protein.the var2csa protein has been closely associated with pregnancy-associated malaria and is recognized as the main adhesin exposed on the surface of plasmodium falciparum-infected erythrocytes. chondroitin sulfate a was identified as the main host receptor in the placenta. single-domain heavy-chain camelid antibodies, more commonly called nanobodies, were selected and produced against the dbl6ℇ-fcr3 domain of var2csa. crystals of two specific nanobodies, nb2907 and nb2919, identified as strong bind ...201323519802
foot and mouth disease and similar virus infections in camelids: a review.foot and mouth disease (fmd) remains the most important animal disease. the fmd virus is highly contagious and occurs almost exclusively among cloven-hoofed animals such as cattle, sheep, goats, bactrian camels and swine. old world camels (owcs) and new world camels (nwcs) inhabit fmd-endemic countries in south america, north and east africa, and the middle and far east. results of experimental infection of owcs with the virus, and several clinical observations from the field over a century, con ...201223520744
expression of orexin a and its receptor 1 in the epididymis of the south american camelid alpaca (vicugna pacos).orexins a (ox a) and b are two peptides originally discovered in neurons of rat hypothalamus, and later found in different cellular types of the gastrointestinal and genital tracts. they arise from the proteolytic cleavage of a common precursor molecule, prepro-orexin, and bind to two receptors, namely receptor 1 (ox1r) and receptor 2 for orexins, that show different binding affinity. the central role of the two peptides has been extensively studied, whereas their activity in the periphery is st ...201423521689
viral infection modulation and neutralization by camelid nanobodies.lactococcal phages belong to a large family of siphoviridae and infect lactococcus lactis, a gram-positive bacterium used in commercial dairy fermentations. these phages are believed to recognize and bind specifically to pellicle polysaccharides covering the entire bacterium. the phage tp901-1 baseplate, located at the tip of the tail, harbors 18 trimeric receptor binding proteins (rbps) promoting adhesion to a specific lactococcal strain. phage tp901-1 adhesion does not require major conformati ...201323530214
generation of single-domain antibody multimers with three different self-associating peptides.conventional y-shaped antibodies have been widely used in research, diagnostics and therapeutics. their large size result in disadvantages in certain applications, which makes production difficult. different parts of an antibody have been used to replace the whole antibody to make it smaller. single-domain antibodies (sdabs) derived from the camelid heavy chain antibodies are among the smallest antibody fragments engineered for various applications. to improve the affinity of these single- sdabs ...201323538432
v(h)h (nanobody) directed against human glycophorin a: a tool for autologous red cell agglutination assays.the preparation of a v(h)h (nanobody) named ih4 that recognizes human glycophorin a (gpa) is described. ih4 was isolated by screening a library prepared from the lymphocytes of a dromedary immunized by human blood transfusion. phage display and panning against gpa as the immobilized antigen allowed isolating this v(h)h. ih4, representing 67% of the retrieved v(h)h sequences, was expressed as a soluble correctly folded protein in shuffle escherichia coli cells, routinely yielding approximately 10 ...201323541519
nanobodies raised against monomeric α-synuclein distinguish between fibrils at different maturation stages.nanobodies are single-domain fragments of camelid antibodies that are emerging as versatile tools in biotechnology. we describe here the interactions of a specific nanobody, nbsyn87, with the monomeric and fibrillar forms of α-synuclein (αsyn), a 140-residue protein whose aggregation is associated with parkinson's disease. we have characterized these interactions using a range of biophysical techniques, including nuclear magnetic resonance and circular dichroism spectroscopy, isothermal titratio ...201323557833
characterization of hachi (camelus dromedarius) fat extracted from the hump.in this work, the characteristics of fat from the hump of young camels (hachi) were evaluated. the physicochemical properties of the fat were as follows: melting point, 45°c; saponification value, 202.3 mg koh/g oil; refractive index (60°c), 1.468; unsaponifiable matter, 1.37%; free fatty acids (as the percentage of oleic acid), 0.96%; and peroxide value, 3.37 mequiv. o2/kg oil. high-resolution (1)h nuclear magnetic resonance ((1)h nmr) was used for the direct determination of the iodine value o ...201323561157
a novel nanobody against urease activity of helicobacter pylori.helicobacter pylori infection is associated with gastritis and in some cases with gastric and duodenal ulcers, and even adenocarcinoma. antibiotic therapy has significant limitations, such as the high cost and the emergence of antibiotic-resistant strains, generating the need for new treatments. the administration of antibody against h. pylori is a new effective therapeutic strategy. in this study, we successfully developed a single-variable domain of heavy chain antibody against recombinant ure ...201323561799
purification and biochemical characterization of membrane-bound neutral ceramidase from camel brain (camelus dromedarius).ceramidases cleave the n-acyl linkages of ceramide to generate sphingosine and its subsequent product sphingosine-1-phosphate (s1p). ceramide and s1p are important bioactive lipids, and ceramidases are important in regulating the availability of these lipids. in this study, we report the purification and characterization of camel brain neutral ceramidase (cbcdase). the novel cbcdase was purified from camel brain using sequential chromatography of deae-sepharose, phenyl-sepharose, superdex, and m ...201323638321
nanopad: an integrated platform for bacterial production of camel nanobodies aimed at detecting environmental biomarkers.the presence of given antigens in environmental samples (e.g. biodegradative enzymes) reports the quality and catalytic vigor of particular soils or aquatic ecosystems. in this context, we have developed the nanopad system consisting of a complete platform for isolation, amplification, and extracellular production of specific antibodies against antigens that diagnose the occurrence of protein markers in crude environmental samples. the workflow starts with the inoculation of camels (camelus drom ...201323661305
evaluation of methods to improve the diagnosis of systemic inflammation in alpacas.the stoic nature of alpacas and limitations of current diagnostic tests make early recognition of inflammatory diseases in this species challenging.201623662652
gastrointestinal helminths of camels (camelus dromedarius) in center of iran.camels are multipurpose animals in iran. as parasitic diseases are the major cause of impaired meat and milk production in this animal, the present study aimed at determining gastrointestinal helminthic infections of iranian camels in the center of the country. gastrointestinal (gi) tract of 144 carcasses of one-humped camels (camelus dromedarius) slaughtered in yazd, esfahan and kerman provinces' abattoirs were examined for adult helminths. camels were from both sexes and different ages. recove ...201323665708
in vivo neutralization of botulinum neurotoxins serotype e with heavy-chain camelid antibodies (vhh).ingestion of botulinum neurotoxin (bont) results in botulism, a severe and frequent fatal disease known in the world. current treatments rely on antitoxins, such as equine antitoxin and human botulism immunoglobulin. in some cases, side effects have been reported, including early anaphylactic shock and late serum sickness. thus, diagnosis and treatment measure of bont are necessary and crucial. in the present study, a single-domain variable heavy-chain (vhh) antibody fragment was obtained from a ...201323666874
camelid single-domain antibody-fragment engineering for (pre)clinical in vivo molecular imaging applications: adjusting the bullet to its target.molecular imaging is a fast developing field and there is a growing need for specific imaging tracers in the clinic. camelid single-domain antibody-fragments (sdabs) recently emerged as a new class of molecular imaging tracers.201323675652
mapping the conformational space accessible to bace2 using surface mutants and cocrystals with fab fragments, fynomers and xaperones.the aspartic protease bace2 is responsible for the shedding of the transmembrane protein tmem27 from the surface of pancreatic β-cells, which leads to inactivation of the β-cell proliferating activity of tmem27. this role of bace2 in the control of β-cell maintenance suggests bace2 as a drug target for diabetes. inhibition of bace2 has recently been shown to lead to improved control of glucose homeostasis and to increased insulin levels in insulin-resistant mice. bace2 has 52% sequence identity ...201323695257
ultrasonography of the liver and kidneys of healthy camels (camelus dromedarius).this study describes the ultrasonography of the liver and kidneys of healthy camels (camelus dromedarius). images of the liver were obtained from the 11th to 5th intercostal spaces (icss). the distance between the dorsal liver margin and the midline of the back was shortest (39.1 ± 7.4 cm) at the 11th ics and increased cranially to 5th ics. the size of the liver was largest at the 9th ics and smallest at the 5th ics. in 6 camels the right kidney was visualized from the 10th and 11th icss and upp ...201223729824
nanobodies and their potential applications.nanobodies are recombinant, antigen-specific, single-domain, variable fragments of camelid heavy chain-only antibodies. the innate supremacy of nanobodies as a renewable source of affinity reagents, together with their high production yield in a broad variety of expression systems, minimal size, great stability, reversible refolding and outstanding solubility in aqueous solutions, and ability to specifically recognize unique epitopes with subnanomolar affinity, have combined to make them a usefu ...201323730699
prenatal development of the eye tunics in the dromedary camel (camelus dromedarius).studies of ocular development in the dromedary camel (camelus dromedarius) have not been reported previously. the aim of the present investigation was therefore to document the major landmarks and the time course in the prenatal development of the eye tunics in dromedary camel and its accommodation with the surrounding hard environment of the desert. serial histological sections of dromedary camel embryos and foetuses were used. age estimation was made on the basis of gestational size, crown ver ...201423731123
evaluation of four point-of-care glucose meters in alpacas.hospitalized alpacas are often hyperglycemic requiring frequent blood glucose testing.201623734626
recombinant expression of trypanosome surface glycoproteins in pichia pastoris for the diagnosis of trypanosoma evansi infection.serodiagnosis of surra, which causes vast economic losses in livestock, is still based on native antigens purified from bloodstream form trypanosoma (t.) evansi grown in rodents. to avoid the use of laboratory rodents in antigen preparation we expressed fragments of the invariant surface glycoprotein (isg) 75, cloned from t. brucei gambiense cdna, and the variant surface glycoprotein (vsg) rotat 1.2, cloned from t. evansi gdna, recombinantly in pichia (p.) pastoris. the m5 strain of this yeast h ...201323747105
plasma leptin, glucose and non-esterified fatty acid variations in dromedary camels exposed to prolonged periods of underfeeding or dehydration.the involvement of plasma leptin in the adaptation of dromedary camels to harsh conditions such as food or water shortages was studied through 2 experiments. in experiment 1, fourteen female camels were either fed at 68% of maintenance energy requirements (mer) during 112d (n=4) or overfed at 134% of mer during the first 56d and then underfed at 17% of mer the next 56d (ov-un, n=5), or underfed and then overfed for the same durations and energy intake levels (un-ov, n=5). weekly plasma samples s ...201323747566
proteomic characterization and cross species comparison of mammalian seminal plasma.seminal plasma contains a large protein component which has been implicated in the function, transit and survival of spermatozoa within the female reproductive tract. however, the identity of the majority of these proteins remains unknown and a direct comparison between the major domestic mammalian species has yet to be made. as such, the present study characterized and compared the seminal plasma proteomes of cattle, horse, sheep, pig, goat, camel and alpaca. gelc-ms/ms and shotgun proteomic an ...201323748023
characterization of fusobacterium necrophorum isolated from llama and alpaca.fusobacterium necrophorum, a gram-negative, anaerobic bacterium, is an opportunistic animal and human pathogen that causes a variety of infections termed necrobacillosis. there are 2 subspecies of f. necrophorum (subsp. necrophorum and subsp. funduliforme) that differ morphologically and biochemically and in virulence. leukotoxin, a secreted protein, is considered to be the major virulence factor. in camelids, f. necrophorum causes a variety of infections, generally involving the lips, tongue, p ...201323780933
prevalence of mycobacterium avium subsp. paratuberculosis fecal shedding in alpacas presented to veterinary hospitals in the united states.the prevalence of johne's disease in alpacas in the united states is unknown. the limits of polymerase chain reaction (pcr) detection of mycobacterium avium subsp. paratuberculosis (map) in alpaca feces have not been determined.201623782278
first report on circulation of echinococcus ortleppi in the one humped camel (camelus dromedaries), sudan.echinococcus granulosus (eg) complex, the cause of cystic echinococcosis (ce), infects humans and several other animal species worldwide and hence the disease is of public health importance. ten genetic variants, or genotypes designated as (g1-g10), are distributed worldwide based on genetic diversity. the objective of this study was to provide some sequence data and phylogeny of eg isolates recovered from the sudanese one-humped camel (camelus dromedaries). fifty samples of hydatid cysts were c ...201323800362
a re-description of physocephalus dromedarii stat. nov. (nematoda: spirocercidae), an abomasal nematode of dromedaries (camelus dromedarius).physocephalus dromedarii stat. nov. was found in the abomasum of two adult female dromedaries originating from a farm in dubai. previously, this nematode has been misidentified as a subspecies of ph. sexalatus but morphological differences are striking enough for it to be upgraded to a species. physocephalus dromedarii is larger than ph. sexalatus and has longer spicules. there are 20-22 oblique crests at the ventral surface between the caudal alae of the male compared to 7-8 in the case of ph. ...201423827046
ultrasonographic fetometry and prenatal fetal sex assessment in camels (camelus dromedarius).the aims of this study were to determine the developmental patterns of some fetal parts to achieve a high accuracy level in the assessment of gestational age and to assess the feasibility and accuracy of ultrasonic prenatal fetal sex assessment in camels. serial ultrasonographic examinations were carried out on seven pregnant dromedary camels. a total of 329 ultrasonographic examinations were conducted between the second and the 54th weeks of pregnancy. intrauterine fluid accumulation was detect ...201323830233
through the eye of an electrospray needle: mass spectrometric identification of the major peptides and proteins in the milk of the one-humped camel (camelus dromedarius).the milk of the one-humped camel (camelus dromedarius) reportedly offers medicinal benefits, perhaps because of its unique bioactive components. milk proteins were determined by (1) two-dimensional gel electrophoresis and peptide mass mapping and (2) liquid chromatography-tandem mass spectrometry (lc-ms/ms) following one-dimensional polyacrylamide gel electrophoresis. over 200 proteins were identified: some known camel proteins including heavy-chain immunoglobulins and others exhibiting regions ...201323832934
neutralizing nanobodies targeting diverse chemokines effectively inhibit chemokine function.chemokine receptors and their ligands play a prominent role in immune regulation but many have also been implicated in inflammatory diseases such as multiple sclerosis, rheumatoid arthritis, allograft rejection after transplantation, and also in cancer metastasis. most approaches to therapeutically target the chemokine system involve targeting of chemokine receptors with low molecular weight antagonists. here we describe the selection and characterization of an unprecedented large and diverse pa ...201323836909
physiological and behavioral responses to different watering intervals in lactating camels (camelus dromedarius).during drought periods camels are watered at long intervals, but effects on body fluid homeostasis of lactating camels are not known. it was hypothesized that camels store water after drinking and minimize water losses by diurnal variation in body temperature, changes in behavior, and release of vasopressin. the aim was to find a sustainable watering interval for lactating camels. seven lactating camels were studied in a cross-over trial in which they were watered once daily (w1), every fourth d ...201323842680
microbiological quality and somatic cell count in bulk milk of dromedary camels (camelus dromedarius): descriptive statistics, correlations, and factors of variation.the objectives of the present study were to monitor the microbiological quality and somatic cell count (scc) of bulk tank milk at the world's first large-scale camel dairy farm for a 2-yr period, to compare the results of 2 methods for the enumeration of scc, to evaluate correlation among milk quality indicators, and to determine the effect of specific factors (year, season, stage of lactation, and level of production) on milk quality indicators. the study was conducted from january 2008 to janu ...201323849636
pharmacokinetics and distribution of ceftazidime to milk after intravenous and intramuscular administration to lactating female dromedary camels (camelus dromedarius).to determine the plasma disposition kinetics, absolute bioavailability, and milk concentrations of ceftazidime in healthy lactating female dromedary camels (camelus dromedarius) following iv and im administration of a single dose of 10 mg/kg (4.5 mg/lb).201323865886
osseous sequestration in alpacas and llamas: 36 cases (1999-2010).to describe clinical findings, treatments, and outcome in camelids treated for osseous sequestration.201323865887
genome sequences of two pathogenic streptococcus agalactiae isolates from the one-humped camel camelus dromedarius.streptococcus agalactiae causes a range of clinical syndromes in camels (camelus dromedarius). we report the genome sequences of two s. agalactiae isolates that induce abscesses in kenyan camels. these genomes provide novel data on the composition of the s. agalactiae "pan genome" and reveal the presence of multiple genomic islands.201323868134
effects of gestational age on physical findings of immaturity, body weight, and survival in neonatal alpacas (2002-2010).gestational duration is highly variable in camelids (reference range 330-360 days), and a definition for prematurity has not been established. anecdotally, it is suggested that crias born outside of this have increased incidence of physical findings of immaturity (tendon laxity, floppy ears, and unerupted teeth) and need increased intensive support in the neonatal period.201623870011
novel g10p[14] rotavirus strain, northern territory, australia.we identified a genotype g10p[14] rotavirus strain in 5 children and 1 adult with acute gastroenteritis from the northern territory, australia. full genome sequence analysis identified an artiodactyl-like (bovine, ovine, and camelid) g10-p[14]-i2-r2-c2-m2-a11-n2-t6-e2-h3 genome constellation. this finding suggests artiodactyl-to-human transmission and strengthens the need to continue rotavirus strain surveillance.201323876354
affinity maturation of a cdr3-grafted vhh using in silico analysis and surface plasmon resonance.a requirement for advancing antibody-based medicine is the development of proteins that can bind with high affinity to a specific epitope related to a critical protein activity site. as a part of generating such proteins, we have succeeded in creating a binding protein without changing epitope by complementarity-determining region 3 (cdr3) grafting (inoue et al., affinity transfer to a human protein by cdr3 grafting of camelid vhh. protein sci. 20, 1971-1981). however, the affinity of the target ...201323902829
a nanobody binding to non-amyloidogenic regions of the protein human lysozyme enhances partial unfolding but inhibits amyloid fibril formation.we report the effects of the interaction of two camelid antibody fragments, generally called nanobodies, namely cab-hul5 and a stabilized and more aggregation-resistant variant cab-hul5g obtained by protein engineering, on the properties of two amyloidogenic variants of human lysozyme, i56t and d67h, whose deposition in vital organs including the liver, kidney, and spleen is associated with a familial non-neuropathic systemic amyloidosis. both nmr spectroscopy and x-ray crystallographic studies ...201323919586
ultrasonography of the lungs and pleura in healthy camels (camelus dromedarius).this study describes ultrasonography of the lungs and pleura in healthy camels (camelus dromedarius). the different layers of the thoracic wall appeared as narrow bands of variable echogenicity. reverberation artefacts appeared as lines of variable echogenicity that ran parallel to the pulmonary surface medial to the pleura. because of its air content, the pulmonary parenchyma was not visualised in all camels. on the right side, the pulmonary surface was seen in the 5th through the 10th intercos ...201323921343
middle east respiratory syndrome coronavirus neutralising serum antibodies in dromedary camels: a comparative serological study.a new betacoronavirus-middle east respiratory syndrome coronavirus (mers-cov)-has been identified in patients with severe acute respiratory infection. although related viruses infect bats, molecular clock analyses have been unable to identify direct ancestors of mers-cov. anecdotal exposure histories suggest that patients had been in contact with dromedary camels or goats. we investigated possible animal reservoirs of mers-cov by assessing specific serum antibodies in livestock.201323933067
a nanobody-based system using fluorescent proteins as scaffolds for cell-specific gene manipulation.fluorescent proteins are commonly used to label cells across organisms, but the unmodified forms cannot control biological activities. using gfp-binding proteins derived from camelid antibodies, we co-opted gfp as a scaffold for inducing formation of biologically active complexes, developing a library of hybrid transcription factors that control gene expression only in the presence of gfp or its derivatives. the modular design allows for variation in key properties such as dna specificity, trans ...201323953120
use of the single cell gel electrophoresis (comet assay) for comparing apoptotic effect of conventional antibodies versus nanobodies.the large molecular size of antibodies is considered one major factor preventing them from becoming more efficient therapeutically. it is well established that all camelids have unique antibodies circulating in their blood called heavy-chain antibodies (hcabs). unlike antibodies from other species, these hcabs contain a single variable domain and two constant domains (ch2 and ch3). hcabs are a novel type of immunoglobulin-like, antigen binding protein with beneficial pharmacokinetic properties t ...201223960797
effect of age, pubertal stage and season on testosterone concentration in male dromedary camel.the present study was conducted in the laboratory of animal physiology and biotechnology, department of animal production, faculty of agriculture, mansoura university, egypt. the present investigation aimed at studying effects of ages, pubertal stages and seasons of the year on testosterone concentrations in blood plasma and tissue homogenate of the testes. the testes used in the current study were collected from a total of 104 one-humped male camels (camelus dromedarius). samples were taken fro ...201023961082
effect of ovary preservation period on recovery rate and categories of dromedary camel oocytes.the aim of this study was to evaluate the effect of different periods of ovary preservation at 25-30 °c for 5, 6, 7, 9, 12 and 24 h on recovery rate and oocyte categories of dromedary camel oocytes. camel ovaries were collected from el-bassatein slaughterhouse, cairo. the collected ovaries were placed immediately after slaughtering into thermos in saline solution (0.9% nacl) supplemented with antibiotics (100 iu penicillin and 100 μg streptomycin/ml) at 25-30 °c and transported to the laboratory ...201023961083
viral and bacterial infections associated with camel (camelus dromedarius) calf diarrhea in north province, saudi arabia.diarrhea and deaths in new-born camel calves were noticed by veterinary investigators and pastoralist in saudi arabia to be very high. hence, it is thought to be necessary to investigate this problem from the virological and bacteriological point of view. the role of pathogenic bacteria and viruses in six different towns of north province (al-assafia, arar, domat aljandal, hail, skaka and khoa) in saudi arabia was studied. survey was conducted in diarrheic camel calves aged 12 months or younger. ...201123961160
generation of vhh antibodies against the arabidopsis thaliana seed storage proteins.antibodies and antibody derived fragments are excellent tools for the detection and purification of proteins. however, only few antibodies targeting arabidopsis seed proteins are currently available. here, we evaluate the process to make antibody libraries against crude protein extracts and more particularly to generate a vhh phage library against native arabidopsis thaliana seed proteins. after immunising a dromedary with a crude arabidopsis seed extract, we cloned the single-domain antigen-bin ...201423963604
amblyomma parvitarsum (acari: ixodidae): localities, hosts and host-parasite ecology.only a few aspects of the biology of amblyomma parvitarsum neumann are known. adults of this hard tick species are parasites of south american camelids in the andean plateau of argentina, bolivia, peru and chile and also in the argentine patagonia, but they have been also rarely found on other artyodactils and two species of birds. the larva has been collected from reptiles in northern chile, whereas the hosts for the nymph remain unknown. on nine localities included within altitude tropical and ...201423979654
generation of a phage-display library of single-domain camelid vh h antibodies directed against chlamydomonas reinhardtii antigens, and characterization of vh hs binding cell-surface antigens.single-domain antibodies (sdabs) are powerful tools for the detection, quantification, purification and subcellular localization of proteins of interest in biological research. we have generated camelid (lama pacos) heavy chain-only variable vh domain (vh h) libraries against antigens in total cell lysates from chlamydomonas reinhardtii. the sdabs in the sera from immunized animals and vh h antibody domains isolated from the library show specificity to c. reinhardtii and lack of reactivity to an ...201323980604
a transversal study on antibodies against selected pathogens in dromedary camels in the canary islands, spain.the canary islands contain the most important dromedary camel (camelus dromedarius) population in the european union and are the main export point of dromedaries to continental europe and latin america. we investigated the presence of antibodies against relevant disease agents in 100 canarian camel sera. selected blood samples of the same animals were also tested by pcr. sera were tested for antibodies against bluetongue virus (btv; 0%), bovine viral diarrhoea virus (bvdv; 0%), camelpox virus (c ...201323992795
population dynamics of ticks infesting the one-humped camel (camelus dromedarius) in central tunisia.a tick population was monitored on 30 camels (camelus dromedarius) over one year in kairouan region, central tunisia. a total of 1630 ticks was collected and identified resulting in an estimate of different parasitological indicators. the ticks belonged to 2 genera and 5 species: hyalomma impeltatum (53%) and hyalomma dromedarii (45%) were the dominant species followed by hyalomma excavatum (1%), hyalomma marginatum (0.5%), and rhipicephalus turanicus (0.5%) (p<0.001). mean infestation prevalenc ...201323999226
the effect of tick infestation on the serum concentrations of the cardiac biomarker troponin i, acid-base balance and haematobiochemical profiles in camels (camelus dromedarius).the aim of the present study was to evaluate the serum concentration of cardiac troponin i (ctni) in camels with tick infestation as a marker of potential myocardial injury and its prognostication. the effects of acid-base and electrolyte balance and haematobiochemical profiles were also investigated. twenty-three camels (camelus dromedarius) with tick infestation and suffering from anorexia, incoordination of movement, unsteady gait, recumbency, opisthotonus, anaemia and reduced production were ...201424002539
ejaculate fractioning effect on llama sperm head morphometry as assessed by the isas(®) casa system.south american camelid sperm characteristics are poorly known compared with those of other domestic animals. the long-term duration of ejaculation makes difficult to gather all the seminal fluid, implying possible ejaculation portion losses. thus, the aim of this research was to evaluate the characteristics of the morphology and morphometry of the spermatozoa change during ejaculation. the morphometric characterization was tested on nine specimens of the lanuda breed, using a special artificial ...201424007277
molecular genotyping of echinococcus granulosus from dromedaries (camelus dromedarius) in eastern iran.with the aim of genotyping echinococcus granulosus cysts found in iranian dromedaries (camelus dromedarius), 50 cysts of e. granulosus were collected from five geographical regions in iran. cysts were characterized using polymerase chain reaction-restriction fragment length polymorphism (pcr-rflp) analysis of the internal transcribed spacer 1 (its1) gene and sequencing fragments of the genes coding for mitochondrial cytochrome c oxidase subunit 1 (cox1). morphological criteria using rostellar ho ...201524018210
adrenaline-activated structure of β2-adrenoceptor stabilized by an engineered nanobody.g-protein-coupled receptors (gpcrs) are integral membrane proteins that have an essential role in human physiology, yet the molecular processes through which they bind to their endogenous agonists and activate effector proteins remain poorly understood. so far, it has not been possible to capture an active-state gpcr bound to its native neurotransmitter. crystal structures of agonist-bound gpcrs have relied on the use of either exceptionally high-affinity agonists or receptor stabilization by mu ...201324056936
t cells expressing vhh-directed oligoclonal chimeric her2 antigen receptors: towards tumor-directed oligoclonal t cell therapy.adoptive cell therapy with engineered t cells expressing chimeric antigen receptors (cars) originated from antibodies is a promising strategy in cancer immunotherapy. several unsuccessful trials, however, highlight the need for alternative conventional binding domains and the better combination of costimulatory endodomains for car construction to improve the effector functions of the engineered t cells. camelid single-domain antibodies (vhhs), which are the smallest single domain antibodies, can ...201424076235
seroepidemiology for mers coronavirus using microneutralisation and pseudoparticle virus neutralisation assays reveal a high prevalence of antibody in dromedary camels in egypt, june 2013.we describe a novel spike pseudoparticle neutralisation assay (ppnt) for seroepidemiological studies on middle east respiratory syndrome coronavirus (merscov) and apply this assay together with conventional microneutralisation (mn) tests to investigate 1,343 human and 625 animal sera. the sera were collected in egypt as a region adjacent to areas where mers has been described, and in hong kong, china as a control region. sera from dromedary camels had a high prevalence of antibody reactive to me ...201324079378
clotting factor viii (fviii) and thrombin generation in camel plasma: a comparative study with humans.the objective of this study was to characterize the highly elevated levels of clotting factor viii (fviii) in camel plasma. whole blood was collected from healthy camels and factor viii clotting activity (fviii:c) assays were conducted using both the clotting and the chromogenic techniques. the anticoagulant citrate phosphate dextrose adenine (cpda) produced the highest harvest of fviii:c, the level of plasma factor viii, compared to heparin:saline and heparin:cpda anticoagulants. camel fviii ca ...201324082408
performance evaluation of affinity ligands for depletion of abundant plasma proteins.human plasma is a commonly used diagnostic fluid in clinical chemistry. in-depth plasma proteomic analysis is performed to search for disease biomarkers, however the large dynamic range of protein abundance in plasma presents a substantial analytical challenge. removal of abundant plasma proteins using antibody capture approaches is a common and attractive means to reduce sample complexity and to aid the analysis of lower abundance proteins of interest. a novel class of heavy chain camelid-deriv ...201324090752
molecular detection of rickettsia africae, rickettsia aeschlimannii, and rickettsia sibirica mongolitimonae in camels and hyalomma spp. ticks from israel.in this study, we aimed to identify and genetically characterize spotted fever group (sfg) rickettsiae in ticks, domestic one-humped camels, and horses from farms and bedouin communities in southern israel. a total of 618 ixodid ticks (hyalomma dromedarii, hyalomma turanicum, hyalomma excavatum, and hyalomma impeltatum) collected from camels and horses, as well as 152 blood samples from 148 camels and four horses were included in the study. initial screening for rickettsiae was carried out by ta ...201324107206
novel therapy based on camelid nanobodies.nanobodies (nbs) are small antibody fragments derived from camelid heavy chain antibodies through recombinant gene technology. their exceptional physicochemical properties, possibility of humanization and unique antigen recognition properties make them excellent candidates for targeted delivery of biologically active components. several different therapeutic approaches based on the novel camelid nbs have been developed to treat a wide range of diseases ranging from immune, bone, blood and neurol ...201324116915
evaluation of sexual behavior of housed male camels (camelus dromedarius) through female parades: correlation with climatic parameters.camels are seasonal breeders, and their sexual behavior is influenced by environmental conditions, but the relationship between climatic factors and sexual behavior has been poorly described in the available literature. nowadays, the male camel living habit is shifting towards captivity; thus, this study was carried out to evaluate the sexual behavior of housed male dromedary camel through female's parades and to correlate it with climatic parameters. four housed sires, reared for semen collecti ...201424122649
characteristics of residency training associated with first-time pass rate on the american college of veterinary ophthalmologists certifying examination.the aim of this study was to examine the association of various aspects of veterinary ophthalmology residency training with the first-time pass rate (ftpr) of the american college of veterinary ophthalmologists (acvo) examination, as well as the individual written, image recognition, animal examination, and surgical sections of the examination.201424131796
mycobacterium bovis infections in domesticated non-bovine mammalian species. part 2: a review of diagnostic methods.despite the large host range of mycobacterium bovis, ante-mortem diagnostic tests for the infection mostly lack sensitivity/specificity and/or remain unvalidated in non-bovine species. the epidemiology and importance of m. bovis infection in these species are discussed in the first part of this two-part review. this second part focuses on the diagnostic options available to identify infected species such as sheep, goats, dogs, cats, and camelids, and highlights the significant challenges posed, ...201324135547
isolation and characterization of serum albumin from camelus dromedarius.serum albumin constitutes 35-50 mg/ml of plasma proteins and performs various physiological activities including the regulation of osmotic pressure on blood, maintaining buffering of the blood ph, carrying different fatty acids and other small molecules, such as bilirubin, hormones, drugs and metal ions, as well as participating in immunological responses. serum albumin is an extensively used protein in biotechnological and pharmaceutical industries. the camel (camelus dromedarius) is well tailo ...201324137219
contributions of the complementarity determining regions to the thermal stability of a single-domain antibody.single domain antibodies (sdabs) are the recombinantly-expressed variable domain from camelid (or shark) heavy chain only antibodies and provide rugged recognition elements. many sdabs possess excellent affinity and specificity; most refold and are able to bind antigen after thermal denaturation. the sdab a3, specific for the toxin staphylococcal enterotoxin b (seb), shows both sub-nanomolar affinity for its cognate antigen (0.14 nm) and an unusually high melting point of 85°c. understanding the ...201324143255
evaluation of antidiphtheria toxin nanobodies.nanobodies are the smallest fragments of naturally occurring single-domain antibodies that have evolved to be fully functional in the absence of a light chain. conventional antibodies are glycoproteins comprising two heavy and two light chains. surprisingly, all members of the camelidae family possess a fraction of antibodies devoid of both light chains and the first constant domain. these types of antibodies are known as heavy-chain antibody (hcab) nanobodies. there are three subclasses of igg ...201024198468
stepwise engineering of heterodimeric single domain camelid vhh antibodies that passively protect mice from ricin toxin.in an effort to engineer countermeasures for the category b toxin ricin, we produced and characterized a collection of epitopic tagged, heavy chain-only antibody vh domains (vhhs) specific for the ricin enzymatic (rta) and binding (rtb) subunits. among the 20 unique ricin-specific vhhs we identified, six had toxin-neutralizing activity: five specific for rta and one specific for rtb. three neutralizing rta-specific vhhs were each linked via a short peptide spacer to the sole neutralizing anti-rt ...201324202178
l-plastin nanobodies perturb matrix degradation, podosome formation, stability and lifetime in thp-1 macrophages.podosomes are cellular structures acting as degradation 'hot-spots' in monocytic cells. they appear as dot-like structures at the ventral cell surface, enriched in f-actin and actin regulators, including gelsolin and l-plastin. gelsolin is an ubiquitous severing and capping protein, whereas l-plastin is a leukocyte-specific actin bundling protein. the presence of the capping protein capg in podosomes has not yet been investigated. we used an innovative approach to investigate the role of these p ...201324236012
effects of dehydration and blockade of angiotensin ii at1 receptor on stress hormones and anti-oxidants in the one-humped camel.the objective of this study was to provide for the first time data on plasma catecholamines, cortisol, glutathione and malondialdehyde after long term dehydration (20 days) in the presence and absence of angiotensin ii (ang ii) at1 receptor blocker (losartan) versus levels in time-matched, non-dehydrated control camels and to record the responses of glutathione and malondialdehyde activity in liver and kidney homogenates in control, dehydrated-losartan treated and dehydrated camels. eighteen mal ...201324252635
activation and allosteric modulation of a muscarinic acetylcholine receptor.despite recent advances in crystallography and the availability of g-protein-coupled receptor (gpcr) structures, little is known about the mechanism of their activation process, as only the β2 adrenergic receptor (β2ar) and rhodopsin have been crystallized in fully active conformations. here we report the structure of an agonist-bound, active state of the human m2 muscarinic acetylcholine receptor stabilized by a g-protein mimetic camelid antibody fragment isolated by conformational selection us ...201324256733
unveiling current guanaco distribution in chile based upon niche structure of phylogeographic lineages: andean puna to subpolar forests.niche description and differentiation at broad geographic scales have been recent major topics in ecology and evolution. describing the environmental niche structure of sister taxa with known evolutionary trajectories stands out as a useful exercise in understanding niche requirements. here we model the environmental niche structure and distribution of the recently resolved phylogeography of guanaco (lama guanicoe) lineages on the western slope of the southern andes. using a maximum entropy fram ...201324265726
seasonal plasticity of the pituitary pars intermedia of the one-humped camel (camelus dromedarius).the pituitary pars intermedia of camelus dromedarius is well developed and completely surrounds the pars nervosa. two major groups of cells are present: endocrine (ec) and glial-like cells (glc). the ec group is composed of three morphologically distinct cell types. type i, or polyhedral light cells (lc-i) and type ii, or polyhedral dark cells (dc-ii), have secretory granules of heterogeneous electron density whose size ranges from 170 to 300nm. type iii cells are elongated with homogeneous elec ...201424295886
identification and in vitro characterization of phage-displayed vhhs targeting vegf.vascular endothelial growth factor (vegf) is a potential target for cancer treatment because of its role in angiogenesis and its overexpression in most human cancers. currently, anti-vegf antibodies have been shown to be promising tools for therapeutic applications. however, large size, poor tumor penetration, immunogenicity, and production in cost- and labor-intensive conditions are major drawbacks of such agents. the antigen-binding regions of camelid single-chain antibodies (vhhs), due to the ...201424297044
structural basis for antibody recognition in the receptor-binding domains of toxins a and b from clostridium difficile.clostridium difficile infection is a serious and highly prevalent nosocomial disease in which the two large, rho-glucosylating toxins tcda and tcdb are the main virulence factors. we report for the first time crystal structures revealing how neutralizing and non-neutralizing single-domain antibodies (sdabs) recognize the receptor-binding domains (rbds) of tcda and tcdb. surprisingly, the complexes formed by two neutralizing antibodies recognizing tcda do not show direct interference with the pre ...201424311789
regulation of β2-adrenergic receptor function by conformationally selective single-domain intrabodies.the biologic activity induced by ligand binding to orthosteric or allosteric sites on a g protein-coupled receptor (gpcr) is mediated by stabilization of specific receptor conformations. in the case of the β2 adrenergic receptor, these ligands are generally small-molecule agonists or antagonists. however, a monomeric single-domain antibody (nanobody) from the camelid family was recently found to allosterically bind and stabilize an active conformation of the β2-adrenergic receptor (β2ar). here, ...201424319111
retinal ganglion cell distribution and visual acuity in alpacas (vicugna pacos).to investigate the distribution of retinal ganglion cells (rgcs) and visual acuity in alpacas (vicugna pacos) through brn-3a immunofluorescent labeling.201524320877
ultrasonographic characterization of follicle deviation in follicular waves with single dominant and codominant follicles in dromedary camels (camelus dromedarius).follicular wave emergence was synchronized by treating camels with gnrh when a dominant follicle (df) was present in the ovaries. animals were scanned twice a day from day 0 (day of gnrh treatment) to day 10, to characterize emergence and deviation of follicles during the development of the follicular wave. follicle deviation in individual animals was determined by graphical method. single dfs were found in 16, double dfs in 9 and triple dfs in two camels. the incidence of codominant (double and ...201424325149
feasibility of utilising an infrared-thermographic technique for early detection of subclinical mastitis in dairy camels (camelus dromedarius).despite the proven ability of infrared thermography (irt) technology for early detection of subclinical mastitis in dairy cows/sheep/goats, studies on its diagnostic feasibility in dairy camels are lacking. sixty-five lactating camels in mid lactation, machine milked twice-daily and managed under intensive conditions were screened to evaluate the feasibility of utilising irt compared with other routine indicators in detecting subclinical mastitis. immediately before the morning milking, a portab ...201424331516
chemical composition and structural characteristics of arabian camel (camelus dromedarius) m. longissimus thoracis.saudi arabian camels of four breeds (6 animals per breed) were used to evaluate characteristics and quality of their meat. chemical composition, fibre cross sectional area, collagen content, muscle metabolism, cooking loss, ph at 24 h post mortem, colour values (except redness) and shear force of longissimus thoracis (lt) muscle did not differ between the breeds. elevated ph values and short sarcomeres reduced overall tenderisation, with a difference between myofibril fragmentation index (p<0.00 ...201424334045
middle east respiratory syndrome coronavirus (mers-cov) serology in major livestock species in an affected region in jordan, june to september 2013.between june and september 2013, sera from 11 dromedary camels, 150 goats, 126 sheep and 91 cows were collected in jordan, where the first human middle-east respiratory syndrome (mers) cluster appeared in 2012. all sera were tested for mers-coronavirus (mers-cov) specific antibodies by protein microarray with confirmation by virus neutralisation. neutralising antibodies were found in all camel sera while sera from goats and cattle tested negative. although six sheep sera reacted with mers-cov an ...201324342516
middle east respiratory syndrome (mers) coronavirus seroprevalence in domestic livestock in saudi arabia, 2010 to 2013.in saudi arabia, including regions of riyadh and al ahsa, pseudoparticle neutralisation (ppnt) and microneutralisation (mnt) tests detected no antibodies to middle east respiratory syndrome coronavirus (mers-cov) in sheep (n= 100), goats (n= 45), cattle (n= 50) and chickens (n= 240). dromedary camels however, had a high prevalence of mers-cov antibodies. bovine coronavirus (bcov) infected sera from cattle had no cross-reactivity in mers-cov ppnt or mnt, while many dromedary camels’ sera reacted ...201324342517
camelus dromedarius brucellosis and its public health associated risks in the afar national regional state in northeastern ethiopia.a cross-sectional study was carried out in four districts of the afar region in ethiopia to determine the prevalence of brucellosis in camels, and to identify risky practices that would facilitate the transmission of zoonoses to humans. this study involved testing 461 camels and interviewing 120 livestock owners. the modified rose bengal plate test (mrbpt) and complement fixation test (cft) were used as screening and confirmatory tests, respectively. spss 16 was used to analyze the overall preva ...201324344729
middle east respiratory syndrome coronavirus in dromedary camels: an outbreak investigation.middle east respiratory syndrome coronavirus (mers-cov) causes severe lower respiratory tract infection in people. previous studies suggested dromedary camels were a reservoir for this virus. we tested for the presence of mers-cov in dromedary camels from a farm in qatar linked to two human cases of the infection in october, 2013.201424355866
identification of mers-cov in dromedary camels. 201424355867
[first serological study of the prevalence of anaplasma phagocytophilum in dromedary (camelus dromedarius) in tunisia].the aim of the study was to evaluate the seroprevalence of anaplasma phagocytophilum in dromedary (camelus dromedarius). sera of 226 healthy dromedaries from three regions of tunisia (sidi bouzid, bouficha and douz) were tested by indirect immunofluorescence (ifa). the overall infection rate was estimated at 29.2%. the study of risk factors showed that region, age, gender, presence of ticks and types of breeding had no influence on the seroprevalence of a. phagocytophilum. this study indicates f ...201424363017
retroviral transduction of fluonanobody and the variable domain of camelid heavy-chain antibodies to chicken embryonic cells.single domain antibodies from camel heavy chain antibodies (vhh or nanobody), are advantages due to higher solubility, stability, high homology with human antibody, lower immunogenicity and low molecular weight. these criteria make them candidates for production of engineered antibody fragments particularly in transgenic animals.201324375066
computed tomographic and radiographic examination of dental structures in south american camelid specimen of different ages.tooth root problems and periodontal diseases are common in south american camelids (sac). the objective was to evaluate and optimize the imaging technique for dental radiography in sac and to describe the radiographic and computed tomographic (ct) anatomy of normal teeth at different ages. in this study, the heads of 20 healthy sac slaughtered for meat production or euthanized for reasons not related to dental problems included 7 female and 10 male llamas and 3 male alpacas. using a standardized ...201424393365
identification of lamanema chavezi becklund 1963 infection in a llama (lama glama) in the united states.infection with lamanema chavezi, a parasitic nematode of new world camelids, was diagnosed by examination of feces and formalin-fixed liver from a 14-month-old female llama (lama glama) that died after a 6-week illness. infection with l. chavezi was initially suspected when a granuloma containing an unidentified nematode was detected microscopically in the hepatic parenchyma from a necropsy specimen. the subsequent diagnosis of l. chavezi infection was based on the morphologic features of 2 imma ...201424398908
a general strategy for antibody library screening via conversion of transient target binding into permanent reporter deposition.we report here a generally applicable method for the selective covalent attachment of a reporter molecule to a replicating entity that allows one to obtain specific binders from a single round of library screening. we show that selective biotinylation of phage particles displaying a binder to any given target can be achieved by application of a coupled enzyme reaction on the surface of the target-binding phage particles that includes a peroxidase, an oxidase and a catalase. due to the covalent l ...201424402333
stratifying fascin and cortactin function in invadopodium formation using inhibitory nanobodies and targeted subcellular delocalization.invadopodia are actin-rich protrusions arising through the orchestrated regulation of precursor assembly, stabilization, and maturation, endowing cancer cells with invasive properties. using nanobodies (antigen-binding domains of camelid heavy-chain antibodies) as perturbators of intracellular functions and/or protein domains at the level of the endogenous protein, we examined the specific contribution of fascin and cortactin during invadopodium formation in mda-mb-231 breast and pc-3 prostate c ...201424414419
use of fourier-transform infrared spectroscopy to quantify immunoglobulin g concentrations in alpaca serum.rapid, economical, and quantitative assays for measurement of camelid serum immunoglobulin g (igg) are limited. in camelids, failure of transfer of maternal immunoglobulins has a reported prevalence of up to 20.5%. an accurate method for quantifying serum igg concentrations is required.201724417433
lipid profile and thyroid hormone status in the last trimester of pregnancy in single-humped camels (camelus dromedarius).changes in lipid metabolism have been shown to occur during pregnancy. the thyroid hormones affect lipid metabolism. the present study was carried out to find out whether the last trimester of pregnancy affects thyroid hormones, thyroid-stimulating hormone (tsh), lipid, and lipoprotein profile in healthy dromedary camels. twenty clinical healthy dromedary camels aged between 4-5 years were divided into two equal groups: (1) pregnant camels in their last trimester of pregnancy and (2) non-pregnan ...201424429808
prevalence and distribution patterns of sarcocystis in camels (camelus dromedarius) in yazd province, iran.sarcocystis spp., are zoonotic cyst-forming coccidian parasites that cause sarcocystosis. this study was conducted to investigate the prevalence as well as distribution patterns of sarcocystis spp. infection in slaughtered one-humped camels of yazd province, iran. muscles of 130 camels were investigated for either macroscopic or microscopic sarcocysts during summer 2009. no macroscopic cyst was observed in the animals at naked eye inspection. of examined camels, 67 (51.5 %) were positive for bra ...201324431561
common gastrointestinal parasites of indigenous camels (camelus dromedarius) with traditional husbandry management (free-ranging system) in central deserts of iran.gastrointestinal parasites of domesticated animals, especially in animals with traditional husbandry management cause reduction in production and performance of them. aim of our study was to determine prevalence of gastrointestinal parasites in indigenous camels, with traditional husbandry management that have no direct contact with other livestock. this study was carried out between kerman and yazd provinces of iran, midsummer 2011. fecal samples of 100 randomly selected camels of all age and s ...201324431575
analysis of heavy and light chain sequences of conventional camelid antibodies from camelus dromedarius and camelus bactrianus species.camel antibodies have been widely investigated, but work has focused upon the unique heavy chain antibodies found across camelid species. these are homodimers, devoid of light chains and the first constant heavy chain domain. camelid species also display conventional hetero-tetrameric antibodies with identical pairs of heavy and light chains; in camelus dromedarius these constitute 25% of circulating antibodies. few investigations have been made on this subset of antibodies and complete conventi ...201424444705
middle east respiratory syndrome coronavirus antibody reactors among camels in dubai, united arab emirates, in 2005.we tested, using a low starting dilution, sequential serum samples from dromedary camels, sheep and horses collected in dubai from february/april to october of 2005 and from dromedary camels for export/import testing between canada and usa in 2000-2001. using a standard middle east respiratory syndrome coronavirus (mers-cov) neutralization test, serial sera from three sheep and three horses were all negative while sera from 9 of 11 dromedary camels from dubai were positive for antibodies support ...201424456414
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