Septicemia and meningoencephalitis caused by Listeria monocytogenes in two neonatal llamas.

PubMed

Hawkins, Ian K; Ilha, Marcia; Anis, Eman; Wilkes, Rebecca P

2017-09-01

Listeriosis is a disease of humans and domestic mammals (mainly ruminants) with variable manifestations, primarily encephalitis, septicemia, and abortion. Although Listeria monocytogenes readily causes illness in ruminants, the prevalence among domestic South American camelids (llamas and alpacas) is low and has not been documented in their wild counterparts, the vicuna and guanaco. We describe herein the clinical signs, autopsy findings, and histopathology of septicemia and suppurative meningoencephalitis caused by L. monocytogenes in 2 neonatal llamas ( Llama glama) from the same herd. L. monocytogenes was isolated in pure culture and identified by real-time PCR on fresh and paraffin-embedded tissue samples of the brain from both crias. This presentation of septicemic listeriosis with meningoencephalitis in 2 animals from the same group is unusual, especially among llamas.

Urinary indices in llamas fed different diets.

PubMed

Lackey, M N; Belknap, E B; Salman, M D; Tinguely, L; Johnson, L W

1995-07-01

Indices of renal function and damage were measured in 12 healthy male adult llamas fed a diet of mixed alfalfa/grass hay (mixed hay) and water ad libitum. Using a collection bag fitted over the preputial area, urine samples were collected at 6, 12, and 24 hours. Serum samples were obtained concurrently to determine endogenous creatinine clearance (CL), total (TE) and fractional excretion (FE) of electrolytes (Na, K, Cl, P), electrolyte CL, urine and serum osmolality, urine enzyme activities (gamma-glutamyltransferase and N-acetyl-beta-D-glucosaminidase), and urine protein concentration. Urine production was quantified. Three months later, 10 of the 12 llamas were fed a grass hay diet and water ad libitum. Similar samples were obtained, and similar measurements were made. Urine production was higher when the llamas were fed the mixed hay diet. Total urine volume for llamas fed mixed hay ranged from 628 to 1,760 ml/24 h, with a median of 1,307.5 ml/24h, compared with a range of 620 to 1,380 ml/24 h and a median of 927.50 ml/24h for llamas fed grass hay. Median urine osmolality was higher in llamas fed mixed hay (1,906 mOsm/kg of body weight, with a range of 1,237 to 2,529 mOsm/kg), compared with llamas fed grass hay (1,666 mOsm/kg with a range of 1,163 to 2,044 mOsm/kg). Creatinine CL did not vary significantly over time for either diet.(ABSTRACT TRUNCATED AT 250 WORDS)

Ejaculatory process and related semen characteristics.

PubMed

Bravo, P W; Moscoso, R; Alarcon, V; Ordoñez, C

2002-01-01

South American camelids are dribble ejaculators, and urethral contractions occur throughout copulation, which may last 25 min. The urethral contractions and their association with semen characteristics during copulation were determined in llamas and alpacas. A transrectal probe was held in the rectum of the male while copulating an artificial vagina, which was accessed underneath the dummy through a hole. The semen-collecting tube was changed every 5 min. Semen characteristics, color, volume, consistency, motility, concentration, and percentage of live sperm were determined at 5-min intervals. Urethral contractions were evenly distributed during copulation: 40 in alpacas and 63 in llamas (p < .05), with a general range of 11 to 132. Semen color was milky in 63%, and translucent in 36.5% for alpacas; and creamy (9.9%), milky (47%), and translucent (42%) for llamas. The mean volume of ejaculate was 0.3, 0.4, 0.6, 0.7, 0.6, 0.8, 0.3, and 3.0 mL for 5, 10, 15, 20, 25, and 30 min, respectively. Semen consistency was variable: viscous (65%) and semiviscous (34%) in alpacas; and viscous (57%) and semiviscous (42%) in llamas. Spermatic motility varied between 60 and 80% for the llama, and 40 and 80% for the alpaca. Spermatic concentration varied between 60 and 188 x 10(3)/mm3 in llamas, and 30 and 170 x 10(3)/mm3 in alpacas. Percentage of live sperm varied the least: 81 to 90% in llamas and 65 to 90% in alpacas. The ejaculate of llamas and alpacas is not fractionated, urethral contractions are evenly distributed, during copulation, and semen characteristics are present throughout the copulatory period.

The role of nitric oxide in the cardiopulmonary response to hypoxia in highland and lowland newborn llamas.

PubMed

Reyes, Roberto V; Díaz, Marcela; Ebensperger, Germán; Herrera, Emilio A; Quezada, Sebastián A; Hernandez, Ismael; Sanhueza, Emilia M; Parer, Julian T; Giussani, Dino A; Llanos, Aníbal J

2018-01-25

Perinatal hypoxia causes pulmonary hypertension in neonates, including humans. However, in species adapted to hypoxia, such as the llama, there is protection against pulmonary hypertension. Nitric oxide (NO) is a vasodilatator with an established role in the cardiopulmonary system of many species, but its function in the hypoxic pulmonary vasoconstrictor response in the newborn llama is unknown. Therefore, we studied the role of NO in the cardiopulmonary responses to acute hypoxia in high- and lowland newborn llamas. We show that high- compared to lowland newborn llamas have a reduced pulmonary vasoconstrictor response to acute hypoxia. Protection against excessive pulmonary vasoconstriction in the highland llama is mediated via enhancement of NO pathways, including increased MYPT1 and reduced ROCK expression as well as Ca 2+ desensitization. Blunting of pulmonary hypertensive responses to hypoxia through enhanced NO pathways may be an adaptive mechanism to withstand life at high altitude in the newborn llama. Llamas are born in the Alto Andino with protection against pulmonary hypertension. The physiology underlying protection against pulmonary vasoconstrictor responses to acute hypoxia in highland species is unknown. We determined the role of nitric oxide (NO) in the cardiopulmonary responses to acute hypoxia in high- and lowland newborn llamas. The cardiopulmonary function of newborn llamas born at low (580 m) or high altitude (3600 m) was studied under acute hypoxia, with and without NO blockade. In pulmonary arteries, we measured the reactivity to potassium and sodium nitroprusside (SNP), and in lung we determined the content of cGMP and the expression of the NO-related proteins: BKCa, PDE5, PSer92-PDE5, PKG-1, ROCK1 and 2, MYPT1, PSer695-MYPT1, PThr696-MYPT1, MLC20 and PSer19-MLC20. Pulmonary vascular remodelling was evaluated by morphometry and based on α-actin expression. High- compared to lowland newborn llamas showed lower in vivo pulmonary arterial pressor responses to acute hypoxia. This protection involved enhanced NO function, as NO blockade reverted the effect and the pulmonary arterial dilatator response to SNP was significantly enhanced in highland neonates. The pulmonary expression of ROCK2 and the phosphorylation of MLC20 were lower in high-altitude llamas. Conversely, MYPT1 was up-regulated whilst PSer695-MYPT1 and PThr695-MYPT1 did not change. Enhanced NO-dependent mechanisms were insufficient to prevent pulmonary arterial remodelling. Combined, the data strongly support that in the highland newborn llama reduced ROCK, increased MYPT1 expression and Ca 2+ desensitization in pulmonary tissue allow an enhanced NO biology to limit hypoxic pulmonary constrictor responses. Blunting of hypoxic pulmonary hypertensive responses may be an adaptive mechanism to life at high altitude. © 2018 The Authors. The Journal of Physiology © 2018 The Physiological Society.

Maintenance energy requirement of llamas.

PubMed

Carmean, B R; Johnson, K A; Johnson, D E; Johnson, L W

1992-09-01

Five castrated male llamas (mean body weight, 94 kg) were studied in an energy balance trial to determine maintenance energy requirement of llamas. Llamas were fed a 50% oat hay-50% pelleted concentrate diet (2.43 Mcal of metabolizable energy/kg of diet dry matter) at approximately 1.6% of body weight (BW). An 8-day total collection digestion trial was used to determine fecal and urine energy losses. Heat production and methane emissions were determined via indirect respiration calorimetry measurements on each llama fed at the same level of intake as during the digestion trial and subsequently on days 3 and 4 of a period of nonfeeding. Fecal, urine, and methane energy losses of the llamas fed near-maintenance intake were 32.5, 3.5, and 7.1% of gross energy intake, respectively. The postabsorptive metabolic rate, commonly called nonfed (fasting) heat production, was 59.3 kcal/BW0.75. Using a linear relation between postabsorptive and maintenance energy requirement and efficiency of energy use below maintenance of 0.702, metabolizable energy requirement at maintenance was determined to be 84.5 kcal/BW0.75.

Meat quality, proximate composition and muscle fatty acid profile of young llamas (Lama glama) supplemented with hay or concentrate during the dry season.

PubMed

Mamani-Linares, L W; Gallo, C B

2014-01-01

Thirty llamas were used to study the effect of a 90 day feed supplementation on meat quality, chemical composition and muscle fatty acid profile. Treatments were: GR=llama on native pasture until slaughter; GR+SH=like GR, but with overnight free access to barley/alfalfa hay; and GR+SC=like GR, but with overnight free access to a wheat bran/sorghum grain concentrate. The supplementation had no effect on postmortem pH and temperature decline in the Longissimus lumborum muscle (LLM), cooking losses nor Warner-Bratzler shear force values (P>0.05). Meat from GR+SC llama had higher fat content in LLM (P<0.05) compared to GR and GR+SH llama. Intramuscular fat from GR+SH llama showed higher (P<0.01) proportions of polyunsaturated fatty acids, higher (P<0.05) polyunsaturated fatty acids/saturated fatty acids and desirable fatty acids ratio, lower (P<0.05) omega-6/omega-3 (n-6/n-3) ratio, and higher (P<0.01) conjugated linoleic acid. © 2013.

Platelet aggregation responses in clinically healthy adult llamas.

PubMed

Gilbert, Rosanne M; Bird, Karyn E; Kutzler, Michelle A

2009-03-01

Limited information exists regarding hemostasis in camelids despite the importance of platelet function testing in the accurate identification of platelet disorders. As further importation of llamas to North America is restricted, variability in breeding stock will continue to decrease, potentially leading to an increase in heritable bleeding disorders. The objective of this study was to measure platelet aggregation responses in clinically healthy llamas and provide baseline data to which abnormal platelet function may be compared in the future. Blood samples were collected from 39 healthy adult llamas, citrated, and centrifuged to produce platelet-rich plasma (PRP). Within 4 hours of the blood draw, 20 microL of each agonist reagent were added to 180 microL of PRP. Final concentrations of agonists were 2 x 10(-5) M ADP, 0.19 mg collagen/mL PRP, 1 x 10(-4) M epinephrine, and 500 microg arachidonic acid/mL PRP. Llama platelets were most responsive to ADP and collagen, with a maximum percent aggregation (mean+/-SD) of 71.3+/-18.6% and 55.8+/-19% and aggregation rates of 9.5+/-3.9 and 6.7+/-3.7 cm/min, respectively. Llama platelet aggregation in response to epinephrine and arachidonic acid was minimal to absent. This study is the first of its kind to establish baseline values for platelet aggregation in healthy adult llamas.

Seminal Plasma Induces Ovulation in Llamas in the Absence of a Copulatory Stimulus: Role of Nerve Growth Factor as an Ovulation-Inducing Factor

PubMed Central

Berland, Marco A.; Ulloa-Leal, Cesar; Barría, Miguel; Wright, Hollis; Dissen, Gregory A.; Silva, Mauricio E.; Ojeda, Sergio R.

2016-01-01

Llamas are considered to be reflex ovulators. However, semen from these animals is reported to be rich in ovulation-inducing factor(s), one of which has been identified as nerve growth factor (NGF). These findings suggest that ovulation in llamas may be elicited by chemical signals contained in semen instead of being mediated by neural signals. The present study examines this notion. Llamas displaying a preovulatory follicle were assigned to four groups: group 1 received an intrauterine infusion (IUI) of PBS; group 2 received an IUI of seminal plasma; group 3 was mated to a male whose urethra had been surgically diverted (urethrostomized male); and group 4 was mated to an intact male. Ovulation (detected by ultrasonography) occurred only in llamas mated to an intact male or given an IUI of seminal plasma and was preceded by a surge in plasma LH levels initiated within an hour after coitus or IUI. In both ovulatory groups, circulating β-NGF levels increased within 15 minutes after treatment, reaching values that were greater and more sustained in llamas mated with an intact male. These results demonstrate that llamas can be induced to ovulate by seminal plasma in the absence of copulation and that copulation alone cannot elicit ovulation in the absence of seminal plasma. In addition, our results implicate β-NGF as an important mediator of seminal plasma-induced ovulation in llamas because ovulation does not occur if β-NGF levels do not increase in the bloodstream, a change that occurs promptly after copulation with an intact male or IUI of seminal plasma. PMID:27355492

Seminal Plasma Induces Ovulation in Llamas in the Absence of a Copulatory Stimulus: Role of Nerve Growth Factor as an Ovulation-Inducing Factor.

PubMed

Berland, Marco A; Ulloa-Leal, Cesar; Barría, Miguel; Wright, Hollis; Dissen, Gregory A; Silva, Mauricio E; Ojeda, Sergio R; Ratto, Marcelo H

2016-08-01

Llamas are considered to be reflex ovulators. However, semen from these animals is reported to be rich in ovulation-inducing factor(s), one of which has been identified as nerve growth factor (NGF). These findings suggest that ovulation in llamas may be elicited by chemical signals contained in semen instead of being mediated by neural signals. The present study examines this notion. Llamas displaying a preovulatory follicle were assigned to four groups: group 1 received an intrauterine infusion (IUI) of PBS; group 2 received an IUI of seminal plasma; group 3 was mated to a male whose urethra had been surgically diverted (urethrostomized male); and group 4 was mated to an intact male. Ovulation (detected by ultrasonography) occurred only in llamas mated to an intact male or given an IUI of seminal plasma and was preceded by a surge in plasma LH levels initiated within an hour after coitus or IUI. In both ovulatory groups, circulating β-NGF levels increased within 15 minutes after treatment, reaching values that were greater and more sustained in llamas mated with an intact male. These results demonstrate that llamas can be induced to ovulate by seminal plasma in the absence of copulation and that copulation alone cannot elicit ovulation in the absence of seminal plasma. In addition, our results implicate β-NGF as an important mediator of seminal plasma-induced ovulation in llamas because ovulation does not occur if β-NGF levels do not increase in the bloodstream, a change that occurs promptly after copulation with an intact male or IUI of seminal plasma.

Cow, sheep and llama manure at psychrophilic anaerobic co-digestion with low cost tubular digesters in cold climate and high altitude.

PubMed

Martí-Herrero, J; Alvarez, R; Cespedes, R; Rojas, M R; Conde, V; Aliaga, L; Balboa, M; Danov, S

2015-04-01

The aim of this research is to evaluate the co-digestion of cow and llama manure combined with sheep manure, in psychrophilic conditions and real field low cost tubular digesters adapted to cold climate. Four digesters were monitored in cold climate conditions; one fed with cow manure, a second one with llama manure, the third one with co-digestion of cow-sheep manure and the fourth one was fed with llama-sheep manure. The slurry had a mean temperature of 16.6 °C, the organic load rate was 0.44 kgvs m(-3) d(-1) and the hydraulic retention time was 80 days. After one hundred days biogas production was stable, as was the methane content and the pH of the effluent. The co-digestion of cow-sheep manure results in a biogas production increase of 100% compared to the mono-digestion of cow manure, while co-digestion of llama-sheep manure results in a decrease of 50% in biogas production with respect to mono-digestion of llama manure. Copyright © 2015 Elsevier Ltd. All rights reserved.

Grazing behaviour and dry matter intake of llamas (Lama glama) and German black- head mutton sheep (Ovis orientalis forma aries) under Central European conditions.

PubMed

Stölzl, Anna Maria; Lambertz, Christian; Gauly, Matthias

2015-01-01

The aim of the present study was to assess the behaviour of llamas (Lama glama) and German blackhead mutton sheep (Ovis orientalis forma aries) when kept under Central European grazing conditions. In total, six adult female sheep and six adult female llamas were observed by direct observation during one week, in which each group was observed for a total time of 24 h. The animals were kept on the same pasture, but the species were raised in separate plots. Forage height before and after the experimental period were determined using a rising plate meter to calculate the average daily dry matter intake (DMI). Llamas had a daily DMI of 0.85%/BW and sheep of 1.04%/BW, respectively. The following behaviours were recorded by direct observation: grazing standing up, grazing lying down, ruminating standing up, ruminating lying down, lying down, lying down lateral and standing. Both species grazed for more than 50% of the time. Ruminating was predominantly performed while standing and lying by sheep (about 50% of the night and 12% of the day) and while lying by llamas (54% of the night and 10% of the day). In conclusion, sheep and llamas differed in grazing behaviour and daily biorhythm. These differences indicate that sheep and llamas may not synchronize their behaviour when co-grazed, though particularly in co-grazing studies the observation period should be extended.

Pituitary response to repeated copulation and/or gonadotropin-releasing hormone administration in llamas and alpacas.

PubMed

Bravo, P W; Stabenfeldt, G H; Fowler, M E; Lasley, B L

1992-11-01

The response of the pituitary gland and ovary to repeated copulatory periods and/or gonadotropin-releasing hormone (GnRH, i.v. 1000 micrograms) administration was determined in llamas and alpacas. Eighty adult females (41 llamas and 39 alpacas with ovulatory follicles) were divided into three general groups for each species as follows: copulation (one or two copulations at either 6- or 24-h intervals) GnRH treatment (one or two treatments at either 6- or 24-h intervals), and combined treatment (copulation followed by GnRH treatment, or GnRH followed by copulation at either 6- or 24-h intervals). An additional control (nontreated) group was composed of 4 llamas and 4 alpacas. The first copulation or treatment with GnRH provoked LH release sufficient to cause ovulation in most of the females (alpacas, 89%; llamas, 92%); urinary pregnanediol glucuronide values, used to verify ovulation, were significantly elevated 48 h after copulation and/or GnRH treatment. A second stimulus, copulation or GnRH, provoked no LH response with concentrations similar to those in nontreated controls and in females not ovulating. Llamas and alpacas thus were refractory to a second copulatory or GnRH stimulus with regard to LH release for up to 24 h following an initial ovulatory release of LH.

Measurement of milk intake in suckling llamas (Lama glama) using deuterium oxide dilution.

PubMed

Riek, A; Gerken, M; Moors, E

2007-02-01

The objective of the study was to estimate daily milk intake in llama crias and relate nutrient intakes at peak lactation to growth data. Milk intake in 11 suckling llamas was estimated from water kinetics using deuterium oxide (D2O) at d 17, 66, and 128 postpartum. Daily milk intakes averaged 2.6, 2.3, and 2.0 kg at 17, 66, and 128 d postpartum, respectively. Milk intake decreased with age when expressed as daily amount, percentage of body weight (BW), or per kilogram of metabolic size, but the influence of age was eliminated when expressed per gram of daily gain. Because llamas only have one young per parturition, milk intake was equivalent to the daily milk output of the dam, which ranged from 27.6 to 96.9 g/kg of maternal BW(0.75). Compared with different ruminant species, milk production in llamas appears to lie between wild and domestic ruminants used for meat production. Nutrients (dry matter, fat, protein, and lactose) and energy intakes from the milk calculated by combining milk intake and milk composition data decreased with age when expressed as daily amount or per 100 g of BW, but when expressed per gram of daily gain, no clear trend was observed. Maintenance requirement for suckling llamas at peak lactation (17 d postpartum) was 312 kJ of ME/kg of BW(0.83). Combined with milk composition data, the present milk intake estimations at different stages of the lactation can be used to establish recommendations for nutrient and energy requirements of suckling llamas.

Estradiol production by preimplantation blastocysts and increased serum progesterone following estradiol treatment in llamas.

PubMed

Powell, Susan A; Smith, Bradford B; Timm, Karen I; Menino, Alfred R

2007-11-01

Estradiol is a potential candidate for the blastocyst signal responsible for maternal recognition of pregnancy in the llama (Lama glama). Two experiments were conducted to determine if the llama blastocyst produces estradiol during the presumed period of maternal recognition of pregnancy and if exogenous estradiol can extend the luteal phase. In Experiment 1, llamas were superovulated with eCG and mated 7 days later (Day 0=day of mating). Blastocysts were collected nonsurgically on Days 7, 9, or 11 or at necropsy on Days 13 and 15 post-mating and cultured for 48h. Conditioned medium was recovered, replaced with fresh medium at 24-h intervals, and assayed for estradiol-17beta. Estradiol production (pg/blastocyst) over the 48-h culture increased (P<0.05) by day of gestation where more estradiol (P<0.05) was produced by Day 11 compared to Day 7 blastocysts, Day 13 compared to Days 7-11 blastocysts, and Day 15 compared to Days 7-13 blastocysts. A dramatic increase was observed between Days 11 and 13 when estradiol production by Day 13 blastocysts increased (P<0.05) more than 50-fold. In Experiment 2, 30 females were induced to ovulate with hCG (Day 0=day of hCG injection). Starting on Day 7 and continuing through Day 15, animals received daily injections i.m. of 0 (n=11), 5 (n=7), or 10mg (n=12) estradiol benzoate (EB) dissolved in isopropylmyristate. Sera were collected immediately prior to each injection and on Days 16, 17, 18, 20, and 22 and analyzed for progesterone. Progesterone concentrations were greater (P<0.05) on Days 14, 15, 16, and 17 in llamas treated with 10mg EB compared to llamas treated with 0mg EB. These results demonstrate that llama blastocysts produce estradiol and exogenous estradiol can enhance and transiently extend luteal progesterone production. Estradiol produced by the preimplantation llama blastocyst may play a role in maternal recognition of pregnancy and early luteal support.

Meat quality attributes of the Longissimus lumborum muscle of the Kh'ara genotype of llama (Lama glama) reared extensively in northern Chile.

PubMed

Mamani-Linares, L W; Gallo, C B

2013-05-01

Twenty male llama of the Kh'ara genotype, reared extensively in the north of Chile, were slaughtered at ages between 2 and 4 permanent teeth (2 to 3.5years) and analyses were carried out on the Longissimus lumborum muscle, including composition (moisture, fat, protein, ash, cholesterol, amino acids, fatty acid profile and collagen content) and meat quality parameters (pH, color, water holding capacity and Warner-Bratzler shear-force). Llama meat was characterized by a low cholesterol (39.04mg/100g) and intramuscular fat (1.56%) content, a total collagen content of 6.28mg/g, of which 20.28% was soluble collagen. Amino acid composition and fatty acid profile were similar to those found for beef finished on forage. Llama meat showed a low n-6/n-3 (4.69) and hypocholesterolemic/hypercholesterolemic (1.55) ratio and acceptable values of DFA (65.78%). Quality parameters in llama Longissimus muscle were within the ranges reported for more traditional meats such as beef and lamb. Copyright © 2012 Elsevier Ltd. All rights reserved.

Identification of cattle, llama and horse meat by near infrared reflectance or transflectance spectroscopy.

PubMed

Mamani-Linares, L W; Gallo, C; Alomar, D

2012-02-01

Visible and near infrared reflectance spectroscopy (VIS-NIRS) was used to discriminate meat and meat juices from three livestock species. In a first trial, samples of Longissimus lumborum muscle, corresponding to beef (31) llamas (21) and horses (27), were homogenised and their spectra collected in reflectance (NIRSystems 6500 scanning monochromator, in the range of 400-2500 nm). In the second trial, samples of meat juice (same muscle) from the same species (20 beef, 19 llama and 19 horse) were scanned in folded transmission (transflectance). Discriminating models (PLS regression) were developed against "dummy" variables, testing different mathematical treatments of the spectra. Best models indentified the species of almost all samples by their meat (reflectance) or meat juice (transflectance) spectra. A few (three of beef and one of llama, for meat samples; one of beef and one of horse, for juice samples) were classified as uncertain. It is concluded that NIRS is an effective tool to recognise meat and meat juice from beef, llama and horses. Copyright © 2011 Elsevier Ltd. All rights reserved.

Total parenteral nutrition in a neonatal llama.

PubMed

Hovda, L R; McGuirk, S M; Lunn, D P

1990-01-15

Total parenteral nutrition reversed cachexia, dehydration, and electrolyte abnormalities in a neonatal llama suffering from prolonged diarrhea. Complications were not observed during the 8 days that IV-administered fluids and nutritional support were provided.

Role of nitric oxide in in vitro contractile activity of the third compartment of the stomach in llamas.

PubMed

Van Hoogmoed, L; Rakestraw, P C; Snyder, J R; Harmon, F A

1998-09-01

To determine the role of nitric oxide and an apamin-sensitive nonadrenergic-noncholinergic inhibitory transmitter in in vitro contractile activity of the third compartment in llamas. Isolated strips of third compartment of the stomach from 5 llamas. Strips were mounted in tissue baths containing oxygenated Kreb's buffer solution and connected to a polygraph chart recorder to measure contractile activity. Atropine, guanethidine, and indomethacin were added to tissue baths to inhibit muscarinic receptors, adrenoreceptors, and prostaglandin synthesis. Responses to electrical field stimulation following addition of the nitric oxide antagonist Nwo-nitro-L-arginine methyl ester (L-NAME) and apamin were evaluated. Electrical field stimulation (EFS) resulted in a reduction in the amplitude and frequency of contractile activity, followed by rebound contraction when EFS was stopped. Addition of L-NAME resulted in a significant reduction in inhibition of contractile activity. Addition of apamin also resulted in a significant reduction in inhibitory contractile activity at most stimulation frequencies. The combination of L-NAME and apamin resulted in a significant reduction in inhibition at all frequencies. Nitric oxide and a transmitter acting via an apamin-sensitive mechanism appear to be involved in inhibition of contractile activity of the third compartment in llamas. Results suggest that nitric oxide plays an important role in mediating contractile activity of the third compartment in llamas. Use of nitric oxide synthase inhibitors may have a role in the therapeutic management of llamas with lesions of the third compartment.

Lamanema chavezi (Nematoda: Molineidae): epidemiological data of the infection in South American camelids of Northwest Argentina.

PubMed

Cafrune, M M; Marín, R E; Rigalt, F A; Romero, S R; Aguirre, D H

2009-12-23

Faecal samples from llamas (n=708), vicuñas (n=171) and guanacos (n=4) were obtained between December 2004 and May 2009 in three Provinces of Northwest Argentina (Jujuy, Salta and Catamarca) to know the distribution, prevalence and intensity of Lamanema chavezi infection in these South American camelid species (SACs). Faeces were examined by a sedimentation-flotation technique using a Cl(2)Zn+ClNa solution (specific gravity=1.59). Eggs of L. chavezi occurred in 30.3% of 89 llama herds and in 18.5% of 708 llamas sampled with a mean intensity of 271.8 eggs/g (EPG) of faeces (range 20-2120). The highest values for all parameters of the infection were registered in llamas from Catamarca Province. Significant differences (P<0.001, Fisher's exact test) were detected only for the lower prevalence in llamas from Jujuy respect to those from the other two Provinces. The overall individual prevalence of L. chavezi in llamas was lower than in reports from adult domestic camelids of neighbour countries while mean intensity was higher. The individual prevalence of L. chavezi in guanacos was 75.0%, with a mean intensity of 66.0 EPG (range 40-120) while no vicuñas were detected as infected. Most of infected SACs were located at the phytogeographical region of Andean Patagonic Domain with a dispersion ranging between 22 degrees 10' and 26 degrees 40' South latitude.

Urinary and plasma purine derivatives in fed and fasted llamas (Lama glama and L. guanacoe).

PubMed

Bakker, M L; Chen, X B; Kyle, D J; Orskov, E R; Bourke, D A

1996-02-01

The changes in urinary and plasma purine derivatives in response to fasting and level of feeding in llamas were examines. In one experiment, four llamas were gradually deprived of feed within 3 days and then fasted for 6 days. Daily urinary excretion of purine derivatives decreased with feed intake and leveled on the last 3 days of fasting at 177 +/- 26 mumol/kg W0.75. Allantoin and uric acid comprised 71% and 15% of total purine derivatives, respectively, in both fed and fasted states, but hypoxanthine plus xanthine increased from 9% to 36%. Plasma concentration of allantoin declined with feed intake reduction, but those of uric acid (217 mumol/l) and hypoxanthine plus xanthine (27 mumol/l) remained relatively unchanged. Concentration of uric acid was higher than that of allantoin, probably due to a high reabsorption of uric acid in renal tubules, which was measured as over 90%. In a second experiment, the four llamas were fed at 860 and 1740 g dry matter/d in a crossover design. Urinary total purine derivatives excretion responded to feed intake (10.4 vs 14.4 mmol/d), although the observed differences did not reach significance. Compared with some ruminant species, it appears that the llama resembles sheep regarding the magnitude of urinary purine derivatives excretion but is unique in maintaining a high concentration of uric acid in plasma, which could be part of the llama's adaptation to their environment.

Regional brain blood flow and cerebral hemispheric oxygen consumption during acute hypoxaemia in the llama fetus

PubMed Central

Llanos, Aníbal J; Riquelme, Raquel A; Sanhueza, Emilia M; Herrera, Emilio; Cabello, Gertrudis; Giussani, Dino A; Parer, Julian T

2002-01-01

Unlike fetal animals of lowland species, the llama fetus does not increase its cerebral blood flow during an episode of acute hypoxaemia. This study tested the hypothesis that the fetal llama brain maintains cerebral hemispheric O2 consumption by increasing cerebral O2 extraction rather than decreasing cerebral oxygen utilisation during acute hypoxaemia. Six llama fetuses were surgically instrumented under general anaesthesia at 217 days of gestation (term ca 350 days) with vascular and amniotic catheters in order to carry out cardiorespiratory studies. Following a control period of 1 h, the llama fetuses underwent 3 × 20 min episodes of progressive hypoxaemia, induced by maternal inhalational hypoxia. During basal conditions and during each of the 20 min of hypoxaemia, fetal cerebral blood flow was measured with radioactive microspheres, cerebral oxygen extraction was calculated, and fetal cerebral hemispheric O2 consumption was determined by the modified Fick principle. During hypoxaemia, fetal arterial O2 tension and fetal pH decreased progressively from 24 ± 1 to 20 ± 1 Torr and from 7.36 ± 0.01 to 7.33 ± 0.01, respectively, during the first 20 min episode, to 16 ± 1 Torr and 7.25 ± 0.05 during the second 20 min episode and to 14 ± 1 Torr and 7.21 ± 0.04 during the final 20 min episode. Fetal arterial partial pressure of CO2 (Pa,CO2, 42 ± 2 Torr) remained unaltered from baseline throughout the experiment. Fetal cerebral hemispheric blood flow and cerebral hemispheric oxygen extraction were unaltered from baseline during progressive hypoxaemia. In contrast, a progressive fall in fetal cerebral hemispheric oxygen consumption occurred during the hypoxaemic challenge. In conclusion, these data do not support the hypothesis that the fetal llama brain maintains cerebral hemispheric O2 consumption by increasing cerebral hemispheric O2 extraction. Rather, the data show that in the llama fetus, a reduction in cerebral hemispheric metabolism occurs during acute hypoxaemia. PMID:11826180

Fetal brain hypometabolism during prolonged hypoxaemia in the llama

PubMed Central

Ebensperger, Germán; Ebensperger, Renato; Herrera, Emilio A; Riquelme, Raquel A; Sanhueza, Emilia M; Lesage, Florian; Marengo, Juan J; Tejo, Rodrigo I; Llanos, Aníbal J; Reyes, Roberto V

2005-01-01

In this study we looked for additional evidence to support the hypothesis that fetal llama reacts to hypoxaemia with adaptive brain hypometabolism. We determined fetal llama brain temperature, Na+ and K+ channel density and Na+–K+-ATPase activity. Additionally, we looked to see whether there were signs of cell death in the brain cortex of llama fetuses submitted to prolonged hypoxaemia. Ten fetal llamas were instrumented under general anaesthesia to measure pH, arterial blood gases, mean arterial pressure, heart rate, and brain and core temperatures. Measurements were made 1 h before and every hour during 24 h of hypoxaemia (n = 5), which was imposed by reducing maternal inspired oxygen fraction to reach a fetal arterial partial pressure of oxygen (Pa,O2) of about 12 mmHg. A normoxaemic group was the control (n = 5). After 24 h of hypoxaemia, we determined brain cortex Na+–K+-ATPase activity, ouabain binding, and the expression of NaV1.1, NaV1.2, NaV1.3, NaV1.6, TREK1, TRAAK and KATP channels. The lack of brain cortex damage was assessed as poly ADP-ribose polymerase (PARP) proteolysis. We found a mean decrease of 0.56°C in brain cortex temperature during prolonged hypoxaemia, which was accompanied by a 51% decrease in brain cortex Na+–K+-ATPase activity, and by a 44% decrease in protein content of NaV1.1, a voltage-gated Na+ channel. These changes occurred in absence of changes in PARP protein degradation, suggesting that the cell death of the brain was not enhanced in the fetal llama during hypoxaemia. Taken together, these results provide further evidence to support the hypothesis that the fetal llama responds to prolonged hypoxaemia with adaptive brain hypometabolism, partly mediated by decreases in Na+–K+-ATPase activity and expression of NaV channels. PMID:16037083

Evaluation of two platelet-rich plasma processing methods and two platelet-activation techniques for use in llamas and alpacas.

PubMed

Semevolos, Stacy A; Youngblood, Cori D; Grissom, Stephanie K; Gorman, M Elena; Larson, Maureen K

2016-11-01

OBJECTIVE To evaluate 2 processing methods (commercial kit vs conical tube centrifugation) for preparing platelet rich plasma (PRP) for use in llamas and alpacas. SAMPLES Blood samples (30 mL each) aseptically collected from 6 healthy llamas and 6 healthy alpacas. PROCEDURES PRP was prepared from blood samples by use of a commercial kit and by double-step conical tube centrifugation. A CBC was performed for blood and PRP samples. Platelets in PRP samples were activated by means of a freeze-thaw method with or without 23mM CaCl 2 , and concentrations of platelet-derived growth factor-BB and transforming growth factor-β 1 were measured. Values were compared between processing methods and camelid species. RESULTS Blood CBC values for llamas and alpacas were similar. The commercial kit yielded a significantly greater degree of platelet enrichment (mean increase, 8.5 fold vs 2.8 fold) and WBC enrichment (mean increase, 3.7 fold vs 1.9 fold) than did conical tube centrifugation. Llamas had a significantly greater degree of platelet enrichment than alpacas by either processing method. No difference in WBC enrichment was identified between species. Concentrations of both growth factors were significantly greater in PRP samples obtained by use of the commercial kit versus those obtained by conical tube centrifugation. CONCLUSIONS AND CLINICAL RELEVANCE For blood samples from camelids, the commercial kit yielded a PRP product with a higher platelet and WBC concentration than achieved by conical tube centrifugation. Optimal PRP platelet and WBC concentrations for various applications need to be determined for llamas and alpacas.

Genotypic characterization of Chilean llama (Lama glama) and alpaca (Vicugna pacos) pestivirus isolates.

PubMed

Aguirre, I M; Fuentes, R; Celedón, M O

2014-01-31

Llamas and alpacas are domesticated South American camelids (SACs) important to ancestral population in the Altiplano region, and to different communities worldwide where they have been introduced. These ungulates have shown to be susceptible to several livestock viral pathogens such as members of the Pestivirus genus, in particular Bovine Viral Diarrhea (BVDV), but there is little data available on Pestivirus infections in SACs. In this study we aimed to detect and identify Pestivirus genotypes and subgroups infecting SACs in both wild and confined environments. Samples were collected from 136 llamas and 30 alpacas from different areas in the Chilean Altiplano (wild animals), and from 22 llamas and 26 alpacas diagnosed as Pestivirus positive from the Metropolitana region in Chile (confined animals). Seroneutralization tests showed titers lower than 2 in all 166 samples from Chilean Altiplano. These samples were also negative to BVDV isolation, indicating that these animals have not been exposed to Pestivirus. After reactivation of positive samples from the Metropolitana region, the 5' non-codifying region (5'NCR) and E2 glycoprotein were amplified by RT-PCR from the Pestivirus genome. Viral sequences were pairwise compared and phylogenetic trees were constructed. The 5'NCR analysis showed that all 12 sequenced isolates belonged to BVDV-1. Of particular interest, isolates from eight llama and two alpaca were BVDV-1j and two alpacas were BVDV-1b. In agreement with these results, E2 phylogenetic analysis rendered a similar grouping indicating that all 16 isolates belong to BVDV-1. However, the lower availability of E2 sequences determines the creation of a smaller number of sub-groups than the 5'NCR sequences. Based on the E2 sequences, the 5'NCR BVDV 1j group consisting of all the llamas and 3 alpacas are completely included in the E2 BVDV 1e group. Due to the universal availability of the 5'NCR segment, we propose the classification of these Chilean llamas and alpacas Pestivirus isolates as BVDV 1j and BVDV 1b respectively. Thus, this is the first time BVDV-1j is obtained in SACs. In addition, these results indicate Pestivirus infection in llamas and alpacas is associated with bovine population as genotypes and sub-groups are the same as those affecting Chilean livestock. Copyright © 2013 Elsevier B.V. All rights reserved.

[Pregnancy in New World camelids].

PubMed

Gauly, M; Bourke, D

1997-01-01

There is a growing number of llama- and alpacabreeders in Europe. Therefore veterinarians are often asked to intervene in clinical management of different problems, especially reproductive problems. In this review the authors attempt to summarize the different possibilities of pregnancy diagnosis and to give an short overview of reproduction in female llamas and alpacas. South American Camelids are induced ovulators and ovulation will occur within 12 to 24 hours following a single breeding. Llamas and alpacas develop follicular waves. The females will usually not ovulate unless a 7 mm or larger follicle exist on one of the ovaries. Corpus luteum is needed for maintenance of pregnancy.

Evaluation of serum trace mineral, vitamin D, and sex steroid hormone concentration, and survey data in llamas and alpacas with metacarpophalangeal and metatarsophalangeal hyperextension.

PubMed

Semevolos, Stacy A; Reed, Shannon K; Schultz, Loren G

2013-01-01

To characterize serum trace mineral, sex steroid hormone, and vitamin D concentrations and identify factors associated with metacarpophalangeal and metatarsophalangeal hyperextension in llamas and alpacas. Serum samples from 79 llamas and 15 alpacas and owner survey data for 573 llamas and 399 alpacas. Serum samples were stored at -20°C until analysis and were evaluated for trace mineral, vitamin D, estradiol, progesterone, and testosterone concentrations. Information regarding age of onset, number of affected animals in herd, feed and supplements given, type of housing, and management practices was obtained in an owner survey. Higher serum zinc and iron concentrations were associated with metacarpophalangeal and metatarsophalangeal hyperextension in camelids, compared with controls. In summer and fall months, vitamin D concentrations were significantly higher in affected camelids than controls. Overall prevalence was 13.3% in llamas, compared with 0.7% in alpacas. No management factors were found to be predictive of this condition. No other factors examined were associated with metacarpophalangeal and metatarsophalangeal hyperextension. Despite similar supplementation practices and environmental conditions between affected and unaffected animals, an association of high serum zinc, iron, and vitamin D concentrations in affected camelids, compared with controls, may indicate differences of intake or absorption of dietary supplements.

Carotid blood flow changes with behavioral states in the late gestation llama fetus in utero.

PubMed

Blanco, C E; Giussani, D A; Riquelme, R A; Hanson, M A; Llanos, A J

1997-12-19

This study tested the hypothesis that in the llama fetus changes in cerebral blood flow are closely associated with changes in cerebral oxidative metabolism such as occur during transitions between electrocortical states. For the first time reported in any species, instantaneous changes in common carotid blood flow, employed as a continuous index of cerebrovascular perfusion, were related to instantaneous changes in electrocortical activity. Three late gestation fetal llamas were surgically prepared under general anesthesia with vascular catheters, a tracheal and amniotic catheter, and with electrodes implanted to monitor the fetal electrocorticogram (ECoG). In addition, Transonic flow probes were placed around a common carotid artery and a femoral artery. At least 4 days after surgery fetal arterial blood, amniotic and tracheal pressures, carotid and femoral blood flows and the fetal ECoG were recorded continuously. Our results suggest a close association between increases in common carotid blood flow and low voltage ECoG in the llama fetus. Close coupling between instantaneous changes in carotid blood flow and electrocortical states together with the lack of an increase in brain blood flow without increased cerebral oxygen extraction during hypoxemia in the llama fetus supports a fall in cerebral oxidative metabolism in this species during hypoxemic episodes.

Analysis of genetic diversity in Bolivian llama populations using microsatellites.

PubMed

Barreta, J; Gutiérrez-Gil, B; Iñiguez, V; Romero, F; Saavedra, V; Chiri, R; Rodríguez, T; Arranz, J J

2013-08-01

South American camelids (SACs) have a major role in the maintenance and potential future of rural Andean human populations. More than 60% of the 3.7 million llamas living worldwide are found in Bolivia. Due to the lack of studies focusing on genetic diversity in Bolivian llamas, this analysis investigates both the genetic diversity and structure of 12 regional groups of llamas that span the greater part of the range of distribution for this species in Bolivia. The analysis of 42 microsatellite markers in the considered regional groups showed that, in general, there were high levels of polymorphism (a total of 506 detected alleles; average PIC across per marker: 0.66), which are comparable with those reported for other populations of domestic SACs. The estimated diversity parameters indicated that there was high intrapopulational genetic variation (average number of alleles and average expected heterozygosity per marker: 12.04 and 0.68, respectively) and weak genetic differentiation among populations (FST range: 0.003-0.052). In agreement with these estimates, Bolivian llamas showed a weak genetic structure and an intense gene flow between all the studied regional groups, which is due to the exchange of reproductive males between the different flocks. Interestingly, the groups for which the largest pairwise FST estimates were observed, Sud Lípez and Nor Lípez, showed a certain level of genetic differentiation that is probably due to the pattern of geographic isolation and limited communication infrastructures of these southern localities. Overall, the population parameters reported here may serve as a reference when establishing conservation policies that address Bolivian llama populations. © 2012 Blackwell Verlag GmbH.

Diagnostic value of animal-side antibody assays for rapid detection of Mycobacterium bovis or Mycobacterium microti infection in South American camelids.

PubMed

Lyashchenko, Konstantin P; Greenwald, Rena; Esfandiari, Javan; Rhodes, Shelley; Dean, Gillian; de la Rua-Domenech, Ricardo; Meylan, Mireille; Vordermeier, H Martin; Zanolari, Patrik

2011-12-01

Tuberculosis (TB) in South American camelids (SAC) is caused by Mycobacterium bovis or Mycobacterium microti. Two serological methods, rapid testing (RT) and the dual-path platform (DPP) assay, were evaluated using naturally infected SAC. The study population included 156 alpacas and 175 llamas in Great Britain, Switzerland, and the United States. TB due to M. bovis (n = 44) or M. microti (n = 8) in 35 alpacas and 17 llamas was diagnosed by gross pathology examination and culture. Control animals were from herds with no TB history. The RT and the DPP assay showed sensitivities of 71% and 74%, respectively, for alpacas, while the sensitivity for llamas was 77% for both assays. The specificity of the DPP assay (98%) was higher than that of RT (94%) for llamas; the specificities of the two assays were identical (98%) for alpacas. When the two antibody tests were combined, the parallel-testing interpretation (applied when either assay produced a positive result) enhanced the sensitivities of antibody detection to 89% for alpacas and 88% for llamas but at the cost of lower specificities (97% and 93%, respectively), whereas the serial-testing interpretation (applied when both assays produced a positive result) maximized the specificity to 100% for both SAC species, although the sensitivities were 57% for alpacas and 65% for llamas. Over 95% of the animals with evidence of TB failed to produce skin test reactions, thus confirming concerns about the validity of this method for testing SAC. The findings suggest that serological assays may offer a more accurate and practical alternative for antemortem detection of camelid TB.

Effects of nutritional restriction on metabolic, endocrine, and ovarian function in llamas (Lama glama).

PubMed

Norambuena, M C; Silva, M; Urra, F; Ulloa-Leal, C; Fernández, A; Adams, G P; Huanca, W; Ratto, M H

2013-05-01

The objectives of the study were to determine the effects of nutritional restriction on ovarian function in llamas. Mature female llamas were assigned randomly to a Control group, fed 100% of maintenance energy requirements (MER) (n=8), or a Restricted group (n=8) fed from 70% to 40% of MER until a body condition score of 2.5 was attained. Blood samples were taken every-other-day to determine plasma concentrations of LH, estradiol, leptin and metabolic markers, and follicular dynamics were monitored daily by ultrasonography for 30 days (Experiment 1). Llamas were then treated with GnRH to compare the ovulatory response and corpus luteus (CL) development between groups (Experiment 2). Blood samples were taken to measure LH, leptin, progesterone and metabolic markers and ovarian structures were assessed as in Experiment 1. Llamas in the Restricted group had lower body mass and body condition scores than those in the Control group (P<0.001). Plasma concentrations of cholesterol, non-esterified fatty acids, triglycerides, and urea were higher in the Restricted group (P<0.05) than in the Control group. The day-to-day diameter profiles of the dominant follicles were smaller (P<0.05) in the Restricted group than in the Control group but plasma estradiol concentration did not differ. The ovulation rate and LH secretion in response to GnRH did not differ. Day-to-day profiles of CL diameter, plasma progesterone and leptin concentrations were smaller (P<0.01) in the Restricted group. In conclusion, nutritional restriction in llamas was associated with suppressed follicle and CL development, and lower plasma concentrations of progesterone and leptin. Copyright © 2013 Elsevier B.V. All rights reserved.

Diagnostic Value of Animal-Side Antibody Assays for Rapid Detection of Mycobacterium bovis or Mycobacterium microti Infection in South American Camelids▿

PubMed Central

Lyashchenko, Konstantin P.; Greenwald, Rena; Esfandiari, Javan; Rhodes, Shelley; Dean, Gillian; de la Rua-Domenech, Ricardo; Meylan, Mireille; Vordermeier, HMartin; Zanolari, Patrik

2011-01-01

Tuberculosis (TB) in South American camelids (SAC) is caused by Mycobacterium bovis or Mycobacterium microti. Two serological methods, rapid testing (RT) and the dual-path platform (DPP) assay, were evaluated using naturally infected SAC. The study population included 156 alpacas and 175 llamas in Great Britain, Switzerland, and the United States. TB due to M. bovis (n = 44) or M. microti (n = 8) in 35 alpacas and 17 llamas was diagnosed by gross pathology examination and culture. Control animals were from herds with no TB history. The RT and the DPP assay showed sensitivities of 71% and 74%, respectively, for alpacas, while the sensitivity for llamas was 77% for both assays. The specificity of the DPP assay (98%) was higher than that of RT (94%) for llamas; the specificities of the two assays were identical (98%) for alpacas. When the two antibody tests were combined, the parallel-testing interpretation (applied when either assay produced a positive result) enhanced the sensitivities of antibody detection to 89% for alpacas and 88% for llamas but at the cost of lower specificities (97% and 93%, respectively), whereas the serial-testing interpretation (applied when both assays produced a positive result) maximized the specificity to 100% for both SAC species, although the sensitivities were 57% for alpacas and 65% for llamas. Over 95% of the animals with evidence of TB failed to produce skin test reactions, thus confirming concerns about the validity of this method for testing SAC. The findings suggest that serological assays may offer a more accurate and practical alternative for antemortem detection of camelid TB. PMID:22012976

Computed tomographic features of the osseous structures of the external acoustic meatus, tympanic cavity, and tympanic bulla of llamas (Lama glama).

PubMed

Concha-Albornoz, Ismael; Stieger-Vanegas, Susanne M; Cebra, Christopher K

2012-01-01

To evaluate the osseous structures of the external acoustic meatus, tympanic cavity, and tympanic bulla of llamas (Lama glama) by use of computed tomography (CT) and establish measurement values for use in detection of abnormalities associated with the external or middle ear in llamas. 10 adult llama heads without any evidence of ear disease. Heads of 10 healthy llamas euthanized by use of a captive bolt striking the dorsal aspect of the skull were collected. Transverse images of the heads were acquired with 1-mm slice thickness, and images were reconstructed in sagittal and dorsal planes. Measurements of the bony structures of the external and middle ear of each head were obtained. The osseous external acoustic meatus curved ventrally as it tracked medially. Its narrowest portion was located at the level of the tympanic annulus. The tympanic bulla conformation differed widely from the bubble-shaped tympanic bulla in dogs and cats. The bulla was divided by the stylohyoid fossa into a larger caudolateral and a smaller caudomedial process; its interior had a honeycombed structure with pneumatized cells similar to the honeycombed appearance of the human mastoid process. Results provided new information regarding the shape and dimensions of the osseous external and middle ear structures in adult llamas without ear disease. Specific landmarks for location of the external acoustic meatus, tympanic cavity, and tympanic bulla in relation to each other were identified. Knowledge of the CT appearance of normal structures will help clinicians to identify changes attributable to middle ear otitis, external ear canal stenosis, or congenital malformations of the ear in this species.

South American camelid illegal traffic detection by means of molecular markers.

PubMed

Di Rocco, F; Posik, D M; Ripoli, M V; Díaz, S; Maté, M L; Giovambattista, G; Vidal-Rioja, L

2011-11-01

South American camelids comprise the wild species guanaco and vicuña and their respective domestic relatives llama and alpaca. The aim of the present study was to determine by DNA analysis to which of these species belong a herd of camelids confiscated from a llama breeder but alleged to be alpacas by the prosecution, and to evaluate the usefulness of mitochondrial and autosomal DNA markers to solve judicial cases involving camelid taxa. Cytochrome b and cytochrome oxidase I mitochondrial genes and 7 STR were analyzed in 25 confiscated samples. Mitochondrial results were inconclusive because 18 of the sequestered samples presented haplotypes that corresponded to the guanaco haplogroup and the remaining seven belonged to a vicuña linage. Microsatellite data of casework samples and llama reference samples revealed different genetic profiles by the presence of private alleles at two microsatellites suggesting that the confiscated animals could be alpaca, or at least alpaca hybrids instead of pure llama. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

LlamaTags: A Versatile Tool to Image Transcription Factor Dynamics in Live Embryos.

PubMed

Bothma, Jacques P; Norstad, Matthew R; Alamos, Simon; Garcia, Hernan G

2018-06-14

Embryonic cell fates are defined by transcription factors that are rapidly deployed, yet attempts to visualize these factors in vivo often fail because of slow fluorescent protein maturation. Here, we pioneer a protein tag, LlamaTag, which circumvents this maturation limit by binding mature fluorescent proteins, making it possible to visualize transcription factor concentration dynamics in live embryos. Implementing this approach in the fruit fly Drosophila melanogaster, we discovered stochastic bursts in the concentration of transcription factors that are correlated with bursts in transcription. We further used LlamaTags to show that the concentration of protein in a given nucleus heavily depends on transcription of that gene in neighboring nuclei; we speculate that this inter-nuclear signaling is an important mechanism for coordinating gene expression to delineate straight and sharp boundaries of gene expression. Thus, LlamaTags now make it possible to visualize the flow of information along the central dogma in live embryos. Copyright © 2018 Elsevier Inc. All rights reserved.

Cetrorelix suppresses the preovulatory LH surge and ovulation induced by ovulation-inducing factor (OIF) present in llama seminal plasma.

PubMed

Silva, Mauricio E; Smulders, Juan P; Guerra, Monserrat; Valderrama, Ximena P; Letelier, Claudia; Adams, Gregg P; Ratto, Marcelo H

2011-05-30

The purpose of the study was to determine if the effect of llama OIF on LH secretion is mediated by stimulation of the hypothalamus or pituitary gland. Using a 2-by-2 factorial design to examine the effects of OIF vs GnRH with or without a GnRH antagonist, llamas with a growing ovarian follicle greater than or equal to 8 mm were assigned randomly to four groups (n = 7 per group) and a) pre-treated with 1.5 mg of GnRH antagonist (cetrorelix acetate) followed by 1 mg of purified llama OIF, b) pre-treated with 1.5 mg of cetrorelix followed by 50 micrograms of GnRH, c) pre-treated with a placebo (2 ml of saline) followed by 1 mg of purified llama OIF or d) pre-treated with a placebo (2 ml of saline) followed by 50 micrograms of GnRH. Pre-treatment with cetrorelix or saline was given as a single slow intravenous dose 2 hours before intramuscular administration of either GnRH or OIF. Blood samples for LH measurement were taken every 15 minutes from 1.5 hours before to 8 hours after treatment. The ovaries were examined by ultrasonography to detect ovulation and CL formation. Blood samples for progesterone measurement were taken every-other-day from Day 0 (day of treatment) to Day 16. Ovulation rate was not different (P = 0.89) between placebo+GnRH (86%) and placebo+OIF groups (100%); however, no ovulations were detected in llamas pre-treated with cetrorelix. Plasma LH concentrations surged (P < 0.01) after treatment in both placebo+OIF and placebo+GnRH groups, but not in the cetrorelix groups. Maximum plasma LH concentrations and CL diameter profiles did not differ between the placebo-treated groups, but plasma progesterone concentrations were higher (P < 0.05), on days 6, 8 and 12 after treatment, in the OIF- vs GnRH-treated group. Cetrorelix (GnRH antagonist) inhibited the preovulatory LH surge induced by OIF in llamas suggesting that LH secretion is modulated by a direct or indirect effect of OIF on GnRH neurons in the hypothalamus.

Reproductive endocrinology of llamas and alpacas.

PubMed

Bravo, P W

1994-07-01

The physiology of reproduction with emphasis in endocrinology of llamas and alpacas is addressed. Basic concepts of ovarian follicular dynamics, endocrine events associated with induction of ovulation, corpus luteum formation, pregnancy, parturition, postpartum interval, puberty, and sexual behavior on the female are reviewed. Pathologic conditions of the reproductive process are also reviewed.

Peritonitis in a llama caused by Streptococcus equi subsp. zooepidemicus.

PubMed Central

Hewson, J; Cebra, C K

2001-01-01

A 7-month-old, male llama was diagnosed with peritonitis caused by Streptococcus equi subsp. zooepidemicus. Clinical findings, medical treatment, and case outcome are described. Hematogenous dissemination from suspected pneumonia is proposed as the route of infection in this case. Possible transmission of the organism through contact with horses is discussed. PMID:11424579

Ameloblastic odontoma in the mandible of a llama

PubMed Central

Step, Douglas L.; Ritchey, Jerry W.; Drost, William Tod; Bahr, Robert J.

2003-01-01

A 4-year-old llama had an aggressive, multiloculated, expansile bone lesion involving the rostral mandible. The mandibular lesion was imaged using radiography and computed tomography. Antemortem diagnosis of an ossifying fibroma was made histologically. Postmortem findings showed the lesion to be limited to the mandible. Final diagnosis was ameloblastic odontoma. PMID:14601679

Morphofunctional structure of the lingual papillae in three species of South American Camelids: Alpaca, guanaco, and llama.

PubMed

Erdoğan, Serkan; Villar Arias, Silvia; Pérez, William

2016-02-01

The aim of this study was to compare the anatomical and functional characteristics of the lingual papilla among the Camelidae. For this purpose, tongues of alpaca, guanaco, and llama were used. Numerous long and thin filiform papillae were located in the median groove and none were detected on the rest of the dorsal surface of the lingual apex in alpaca. Secondary papillae originated from the base of some filiform papillae on the ventral surface of alpaca tongue. The bases of some filiform papillae of the lateral surface of the lingual apex were inserted into conspicuous grooves in guanaco and tips of filiform papillae on the dorsal surface of the lingual body were ended by bifurcated apex. On the dorsal surface of the lingual apex of llama, there were no filiform papillae but there were numerous filiform papillae on both the lateral margins of the ventral surface of the lingual apex. Fungiform papillae were distributed randomly on dorsal lingual surface and ventral margins of the tongues of all camelid species. Lenticular papillae were located on the lingual torus and varied in size and topographical distribution for each species. Circumvallate papillae had irregular surfaces in llama and alpaca, and smooth surface in guanaco. In conclusion, llama and alpaca tongues were more similar to each other, and tongues of all camelid species displayed more similarities to those of Bactrian and dromedary camels in comparison with other herbivores and ruminants. © 2015 Wiley Periodicals, Inc.

Short communication: milk output in llamas (Lama glama) in relation to energy intake and water turnover measured by an isotope dilution technique.

PubMed

Riek, A; Klinkert, A; Gerken, M; Hummel, J; Moors, E; Südekum, K-H

2013-03-01

Despite the fact that llamas have become increasingly popular as companion and farm animals in both Europe and North America, scientific knowledge on their nutrient requirements is scarce. Compared with other livestock species, relatively little is known especially about the nutrient and energy requirements for lactating llamas. Therefore, we aimed to measure milk output in llama dams using an isotope dilution technique and relate it to energy intakes at different stages of lactation. We also validated the dilution technique by measuring total water turnover (TWT) directly and comparing it with values estimated by the isotope dilution technique. Our study involved 5 lactating llama dams and their suckling young. Milk output and TWT were measured at 4 stages of lactation (wk 3, 10, 18, and 26 postpartum). The method involved the application of the stable hydrogen isotope deuterium ((2)H) to the lactating dam. Drinking water intake and TWT decreased significantly with lactation stage, whether estimated by the isotope dilution technique or calculated from drinking water and water ingested from feeds. In contrast, lactation stage had no effect on dry matter intake, metabolizable energy (ME) intake, or the milk water fraction (i.e., the ratio between milk water excreted and TWT). The ratios between TWT measured and TWT estimated (by isotope dilution) did not differ with lactation stage and were close to 100% in all measurement weeks, indicating that the D(2)O dilution technique estimated TWT with high accuracy and only small variations. Calculating the required ME intakes for lactation from milk output data and gross energy content of milk revealed that, with increasing lactation stage, ME requirements per day for lactation decreased but remained constant per kilogram of milk output. Total measured ME intakes at different stages of lactation were similar to calculated ME intakes from published recommendation models for llamas. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Oestradiol-17β plasma concentrations after intramuscular injection of oestradiol benzoate or oestradiol cypionate in llamas (Lama glama)

PubMed Central

2010-01-01

Background Llamas (Lama glama) are induced ovulators and the process of ovulation depends on dominant follicular size. In addition, a close relationship between behavioural estrus and ovulation is not registered in llamas. Therefore, the exogenous control of follicular development with hormones aims to predict the optimal time to mate. Oestradiol-17β (E2) and its esters are currently used in domestic species, including camelids, in synchronization treatments. But, in llamas, there is no reports regarding the appropriate dosages to be used and most protocols have been designed by extrapolation from those recommended for other ruminants. The aim of the present study was to characterize plasma E2 concentrations in intact female llamas following a single intramuscular (i.m.) injection of two oestradiol esters: oestradiol benzoate (EB) and oestradiol cypionate (ECP). Methods Twelve non pregnant and non lactating sexually mature llamas were i.m. injected on day 0 with 2.5 mg of EB (EB group, n = 6) or ECP (ECP group, n = 6). Blood samples were collected immediately before injection, at 1, 6, 12, 24 h after treatment and then daily until day 14 post injection. Changes in hormone concentrations with time were analyzed in each group by analysis of variance (ANOVA) using a repeated measures (within-SS) design. Plasma E2 concentrations and area under the concentration-time curve (AUC) values were compared between groups by ANOVA. In all cases a Least-Significant Difference test (LSD) was used to determine differences between means. Hormonal and AUC data are expressed as mean ± S.E.M. Results Peak plasma E2 concentrations were achieved earlier and were higher in EB group than in ECP group. Thereafter, E2 returned to physiological concentrations earlier in EB group (day 5) than in ECP group (day 9). Although plasma E2 profiles differed over time among groups there were no differences between them on AUC values. Conclusions The i.m. injection of a single dose of both oestradiol esters resulted in plasma E2 concentrations exceeding physiological values for a variable period. Moreover, the plasma E2 profiles observed depended on the derivative of oestradiol administered. This basic information becomes relevant at defining treatment protocols including oestrogens in llamas. PMID:20149242

Sarcocystis spp. in llamas (Lama glama) in Southern Bolivia: a cross sectional study of the prevalence, risk factors and loss in income caused by carcass downgrades.

PubMed

Rooney, A L; Limon, G; Vides, H; Cortez, A; Guitian, J

2014-10-01

Llamas (Lama glama) are intermediate hosts of the protozoan parasite Sarcocystis spp. This parasite is described as causing economic losses in the production of llama meat in South America. The aim of this study was to estimate prevalence, identify risk factors and explore spatial patterns of Sarcocystis in llamas in an area of the Bolivian High Plateau including estimating financial losses due to carcass downgrades as a result of the presence of Sarcocystis cysts. Information was collected from a local abattoir between 2006 and 2011 on 1196 llamas. Sarcocystis status was determined at meat inspection where any carcasses with one or more visible cysts were deemed Sarcocystis positive. A high prevalence was found, estimated to vary between 23.4% (95% CI 16.6-30.1) in 2007 and 50.3% (95% CI 44.4-56.3) in 2011. Period prevalence between 2006 and 2011 was estimated at 34.1% (95% CI 31.4-36.8). Age, sex and type (analogous to breed) were identified as risk factors for Sarcocystis using a mixed-effects logistic regression model adjusting for clustering by community and owner. Llamas over 4.5 years of age had an increased odds of being Sarcocystis positive (OR 19.31, 95% CI 9.10-40.98) as well as females (OR 1.75, 95% CI 1.13-2.68) and long haired type llamas (OR 1.90, 95% CI 1.26-2.87). An interaction between age and sex was detected indicating that the increased odds of disease from the youngest age group to the 2.5-4.5 years group was much more pronounced in females than in males. Spatial patterns of Sarcocystis were explored at district level by means of Standardised Morbidity Ratios and some spatial heterogeneity was revealed. Estimates of financial loss due to the disease were calculated using the difference in price paid for Sarcocystis positive and negative meat. Loss due to Sarcocystis varied per year but could be up to 20% of the annual income generated through the abattoir by sale of meat. Overall this study shows a high prevalence of Sarcocystis in the study area with some heterogeneity between districts. It also identifies some previously unknown risk factors for Sarcocystis and gives financial estimates of the cost of the disease as a result of carcass downgrades. We hoped these findings will add to the understanding of Sarcocystis in llamas in Southern Bolivia and will be useful when considering if controls are necessary, worthwhile and practical. Copyright © 2013 Elsevier B.V. All rights reserved.

Acute renal failure in 2 adult llamas after exposure to Oak trees (Quercus spp.)

PubMed Central

Chamorro, Manuel F.; Passler, Thomas; Joiner, Kellye; Poppenga, Robert H.; Bayne, Jenna; Walz, Paul H.

2013-01-01

Two adult llamas (Lama glama) previously exposed to oak trees (Quercus spp.) were presented with a history of depression and anorexia. Clinicopathological abnormalities included severe gastroenteritis, acute renal failure, and increased liver enzymes. This is believed to be the first report of oak toxicosis in South American camelids. PMID:23814303

Cutaneous metastases of a mammary carcinoma in a llama.

PubMed Central

Leichner, T L; Turner, O; Mason, G L; Barrington, G M

2001-01-01

An 8-year-old, female llama was evaluated for nonhealing, ulcerative, cutaneous lesions, which also involved the mammary gland. Biopsies of the lesions distant from and within the mammary gland area revealed an aggressive carcinoma. The tumor was confirmed at necropsy to be a mammary gland adenocarcinoma with cutaneous metastasis. Images Figure 1. PMID:11265189

Application of Monoclonal Antibodies in Functional and Comparative Investigations of Heavy-Chain Immunoglobulins in New World Camelids

PubMed Central

Daley, L. P.; Gagliardo, L. F.; Duffy, M. S.; Smith, M. C.; Appleton, J. A.

2005-01-01

Of the three immunoglobulin G (IgG) isotypes described to occur in camelids, IgG2 and IgG3 are distinct in that they do not incorporate light chains. These heavy-chain antibodies (HCAbs) constitute approximately 50% of the IgG in llama serum and as much as 75% of the IgG in camel serum. We have produced isotype-specific mouse monoclonal antibodies (MAbs) in order to investigate the roles of HCAbs in camelid immunity. Seventeen stable hybridomas were cloned, and three MAbs that were specific for epitopes on the γ chains of llama IgG1, IgG2, or IgG3 were characterized in detail. Affinity chromatography revealed that each MAb bound its isotype in solution in llama serum. The antibodies bound to the corresponding alpaca IgGs, to guanaco IgG1 and IgG2, and to camel IgG1. Interestingly, anti-IgG2 MAbs bound three heavy-chain species in llama serum, confirming the presence of three IgG2 subisotypes. Two IgG2 subisotypes were detected in alpaca and guanaco sera. The MAbs detected llama serum IgGs when they were bound to antigen in enzyme-linked immunosorbent assays and were used to discern among isotypes induced during infection with a parasitic nematode. Diseased animals, infected with Parelaphostrongylus tenuis, did not produce antigen-specific HCAbs; rather, they produced the conventional isotype, IgG1, exclusively. Our data document the utility of these MAbs in functional and physiologic investigations of the immune systems of New World camelids. PMID:15753251

Molecular detection of Sarcocystis aucheniae in the blood of llamas from Argentina.

PubMed

Martin, Mara; Decker Franco, Cecilia; Romero, Sandra; Carletti, Tamara; Schnittger, Leonhard; Florin-Christensen, Monica

Sarcocystis aucheniae are apicomplexan protozoa that infect South American camelids (SACs), giving rise to macroscopic cysts similar to rice grains in skeletal muscles. Visual detection of macrocysts in slaughtered animals hampers commercialization of SAC meat, a highly relevant economic exploitation for Andean rural families. Importantly, the consumption of undercooked S. aucheniae-infested meat causes gastroenteritis. A carnivore definitive host, possibly the dog, acquires the parasite when feeding on infected SAC meat, and later eliminates infective oocysts in its feces. The parasite cycle is completed when SACs ingest contaminated water or pastures. We hypothesized that parasite DNA can be detected in SAC blood using molecular methods. In order to test this hypothesis, a seminested PCR format was specifically designed to target the hypervariable 18S rRNA gene region of S. aucheniae. PCR conditions were optimized using genomic DNA extracted from macrocyst bradyzoites. A detection limit of up to 1 parasite in 10μl of llama blood was established based on DNA samples extracted from aliquots of S. aucheniae bradyzoite-spiked non-infected llama blood. The seminested PCR allowed to detect natural infections of S. aucheniae in llama blood samples originating in the Andean flatlands of Argentina. Specific amplification of S. aucheniae DNA was corroborated by amplicon sequencing. This is the first report of S. aucheniae detection in llama blood, which provides a valuable diagnostic tool for epidemiological studies and for the evaluation of the efficacy of control measures for this parasitosis. Copyright © 2016 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

Pathology of Haemonchus contortus in New World camelids in the southeastern United States: a retrospective review.

PubMed

Edwards, Erin E; Garner, Bridget C; Williamson, Lisa H; Storey, Bob E; Sakamoto, Kaori

2016-03-01

Most small ruminant farms in tropical climates are plagued by Haemonchus contortus, a hematophagous, abomasal parasite. Heavy burdens of this parasite can cause anemia, hypoproteinemia, weight loss, and mortality in susceptible animals. Haemonchus contortus is becoming a major health concern in New World camelids as well, namely llamas (Llama glama) and alpacas (Vicugna pacos), yet little research has been conducted regarding its prevalence or pathology in these species. Herein, we present a retrospective review of llamas and alpacas that were admitted to The University of Georgia Veterinary Teaching Hospital and Athens Diagnostic Laboratory between the years 2002 and 2013. Antemortem fecal egg count (FEC) estimates performed on 30 alpacas were negatively correlated with hematocrit, hemoglobin, and red blood cell count. Total protein was not significantly correlated with FEC. On postmortem examination, 55 of 198 camelids, including 2 from the aforementioned antemortem review, were infected with H. contortus, with llamas (42.6%) having a significantly higher infection rate than alpacas (22.2%). In 15.7% of the total cases, the parasite was the major cause of death. Common gross lesions included peritoneal, thoracic, and pericardial effusions, visceral pallor, subcutaneous edema, and serous atrophy of fat. Histologic lesions included centrilobular hepatic necrosis, hepatic atrophy, lymphoplasmacytic inflammation of the mucosa of the third gastric compartment (C3), extramedullary hematopoiesis in both the liver and spleen, and the presence of nematodes in C3. Our study emphasizes the importance of H. contortus diagnosis and herd monitoring in New World camelids, particularly llamas. © 2016 The Author(s).

No detectable carotenoid concentrations in serum of llamas and alpacas.

PubMed

Raila, J; Schweigert, F J; Stanitznig, A; Lambacher, B; Franz, S; Baldermann, S; Wittek, T

2017-08-01

Carotenoids are lipid-soluble pigments and important for a variety of physiological functions. They are major dietary vitamin A precursors and act as lipophilic antioxidants in a variety of tissues and are associated with important health benefits in humans and animals. All animals must acquire carotenoids from their diet, but to our knowledge, there are no studies investigating the intestinal carotenoid absorption and their blood concentrations in New World camelids. The present study aimed to assess the serum concentrations of selected carotenoids in llamas (n = 13) and alpacas (n = 27). Serum carotenoids as well as retinol (vitamin A) and α-tocopherol (vitamin E) were determined by high-performance liquid chromatography coupled with mass spectrometry and these were unable to detect any carotenoids (α- and β-carotene, α- and β-cryptoxanthin, lutein, zeaxanthin, lycopene) in the samples. The concentrations of retinol in alpacas (2.89 ± 1.13 μmol/l; mean ± SD) were higher (p = 0.024) than those found in llamas (2.05 ± 0.87 μmol/l); however, the concentrations of α-tocopherol were not significantly (p = 0.166) different (llamas: 3.98 ± 1.83 μmol/l; alpacas: 4.95 ± 2.14 μmol/l). The results show that both llamas and alpacas are not able to absorb intact carotenoids, but efficiently convert provitamin A carotenoids to retinol. Journal of Animal Physiology and Animal Nutrition © 2016 Blackwell Verlag GmbH.

Assessment of the effects of epinephrine and insulin on plasma and serum biochemical variables in llamas and alpacas.

PubMed

Cebra, Christopher K; Tornquist, Susan J

2004-12-01

To describe the metabolic effects of epinephrine administration in New World camelids and investigate whether these effects are influenced by administration of insulin. 6 llamas and 8 alpacas (all adult castrated males). Prior to each experiment, food was withheld from camelids for 8 hours. On each of 2 consecutive days, alpacas were administered epinephrine (10 mg/kg, IM; time 0); alpacas were randomly assigned to receive regular insulin (0.2 U/kg, IV) immediately after epinephrine administration on one of those days. In llamas, the experiment was performed once after administration of epinephrine only. At 0, 30, 60, 90, 120, 150, 180, 210, and 240 minutes after treatment, blood samples were collected and several serum or plasma biochemical variables were assessed; in addition, plasma samples from llamas were assessed for insulin concentrations. Data were compared between days (alpacas only) and between time points. Administration of epinephrine induced mobilization of glucose, triglycerides, nonesterified fatty acids, and beta-hydroxybutyrate. A small increase in endogenous insulin concentration was detected in epinephrine-treated llamas, compared with baseline values. Overall, insulin administration decreased, negated, or delayed the epinephrine-associated increases in serum or plasma concentrations of circulating energy substrates, except that it augmented the epinephrine-associated increase in concentration of triglycerides. Epinephrine appeared to mobilize energy substrates in camelids and hence may be involved in the pathogenesis of disorders of glucose and fat metabolism. Insulin appeared to antagonize most of these effects, and its administration may have therapeutic value in camelids.

Blood serum concentrations of selenium in female llamas (Lama glama) in relationship to feeding practices, region of United States, reproductive stage, and health of offspring.

PubMed

Herdt, T H

1995-02-01

Serum Se and vitamin E concentrations were determined twice in each of 35 female llamas from a high-Se region and 96 female llamas from a low-Se region of the United States. The first determination was taken at a random time during gestation and the second shortly after parturition. At the time of postpartum sampling, a sample was also taken from the cria. Dietary information was collected from each llama owner and a subjective estimate of strength and vigor was assigned by the owner to each cria studied. The mean blood serum concentrations of Se in llamas during gestation and at parturition were 213 and 203 ng/mL, respectively. The mean serum Se concentration was 113 ng/mL in neonatal cria. Among herds, serum Se means varied for the cria (P < .05) and at both sampling times in the females (P < .01). In adult females, mean serum Se was actually higher during gestation (P = .06) and at parturition (P < .01) for the low-Se region than for the high-Se region, but this effect was removed by covariate adjustment for intake of supplemental Se. Serum Se in females was correlated (P < .05) to supplemental Se consumption. Vigor and (or) viability of the cria were not affected by the Se status of the dams (P > .3) or cria (P > .2). However, there were very few weak or stillborn cria in this study.(ABSTRACT TRUNCATED AT 250 WORDS)

Llama reproduction.

PubMed

Johnson, L W

1989-03-01

The reproductive anatomy, physiology, and breeding behavior of the llama is unique enough to make familiarity with it imperative. Female puberty averages 12 months, while many males are not reproductively functional until after 3 years. Proper management of a breeding pair or herd is necessary to maintain maximum reproductive performance. Proof of pregnancy is suggested by rejection of the male and may/should be confirmed by progesterone assay, rectal palpation, or ultrasound techniques. The postpartum female is notable for breeding back rapidly, with high pregnancy rates resulting. Both male and female factors enter into llama infertility, with each gender having significant incidence of reproductive anatomical abnormalities. Management as well as acquired infertility problems (heat factors, trauma, infection, neoplasia, and hormonal imbalances) contribute to the bulk of infertility cases investigated. Techniques used to diagnose infertility in llamas are quite comparable to the equine species; however, female body size and semen analysis in the male present significant challenges. The approach to therapy has been quite empirical to date, owing to lack of consistent problems and numbers to afford conclusive trials. Alterations of pregnancy include resorption, abortions, and stillbirths. Resorption between 30 to 60 days of gestation is reported regularly. Abortions caused by stress occur regularly. Infection abortions caused by leptospirosis, toxoplasmosis, and chlamydiosis are to be expected. Ponderosa pine-related abortions are suspected. In summary, I find use for a broad background in large animal theriogenology to apply to llama infertility. There no doubt are additional diagnostic techniques and therapeutic regimens that have application, and it is up to us all to keep good records and share the information.

Application of monoclonal antibodies in functional and comparative investigations of heavy-chain immunoglobulins in new world camelids.

PubMed

Daley, L P; Gagliardo, L F; Duffy, M S; Smith, M C; Appleton, J A

2005-03-01

Of the three immunoglobulin G (IgG) isotypes described to occur in camelids, IgG2 and IgG3 are distinct in that they do not incorporate light chains. These heavy-chain antibodies (HCAbs) constitute approximately 50% of the IgG in llama serum and as much as 75% of the IgG in camel serum. We have produced isotype-specific mouse monoclonal antibodies (MAbs) in order to investigate the roles of HCAbs in camelid immunity. Seventeen stable hybridomas were cloned, and three MAbs that were specific for epitopes on the gamma chains of llama IgG1, IgG2, or IgG3 were characterized in detail. Affinity chromatography revealed that each MAb bound its isotype in solution in llama serum. The antibodies bound to the corresponding alpaca IgGs, to guanaco IgG1 and IgG2, and to camel IgG1. Interestingly, anti-IgG2 MAbs bound three heavy-chain species in llama serum, confirming the presence of three IgG2 subisotypes. Two IgG2 subisotypes were detected in alpaca and guanaco sera. The MAbs detected llama serum IgGs when they were bound to antigen in enzyme-linked immunosorbent assays and were used to discern among isotypes induced during infection with a parasitic nematode. Diseased animals, infected with Parelaphostrongylus tenuis, did not produce antigen-specific HCAbs; rather, they produced the conventional isotype, IgG1, exclusively. Our data document the utility of these MAbs in functional and physiologic investigations of the immune systems of New World camelids.

Comparing manager and visitor perceptions of llama use in wilderness

Treesearch

Alan E. Watson; Neal A. Christensen; Dale J. Blahna; Kari S. Archibald

1998-01-01

Llama use in wilderness is projected to increase over the next 5 years. While the greatest concerns about this increase in use are unexpected impacts to native flora, impacts to native fauna, and conflicts with other user types, there is also concern about how prepared managers are to deal with this increasing recreation demand. This research compares manager attitudes...

Infertility associated with persistent hymen in an alpaca and a llama

PubMed Central

Tan, Rachel H.H.; Dascanio, John J.

2008-01-01

Perforation and dilation of the persistent hymen in an alpaca and a llama, detected by vaginal examination and endoscopy, was achieved by use of a sigmoidoscope and incremental dilation using cylindrical instruments to a maximum diameter of 38 mm. Outcome and subsequent fertility are dependent on length of time the obstruction has been present and secondary uterine disease. PMID:19183735

Brainstem auditory evoked responses and ophthalmic findings in llamas and alpacas in eastern Canada

PubMed Central

Cullen, Cheryl L.; Lamont, Leigh A.

2006-01-01

Abstract Seventeen llamas and 23 alpacas of various coat and iris colors were evaluated for: 1) deafness by using brainstem auditory evoked response testing; and 2) for ocular abnormalities via complete ophthalmic examination. No animals were deaf. The most common ocular abnormalities noted were iris-to-iris persistent pupillary membranes and incipient cataracts. PMID:16536233

Prevalence of Mycoplasma haemolamae infection in Peruvian and Chilean llamas and alpacas.

PubMed

Tornquist, Susan J; Boeder, Lisa; Rios-Phillips, Carolina; Alarcon, Virgilio

2010-09-01

Mycoplasma haemolamae is a hemotropic mycoplasma that affects red blood cells of llamas (Lama glama) and alpacas (Lama pacos). It is variably associated with anemia, and most infections are subclinical. Development of a polymerase chain reaction assay has facilitated detection of this infection in llamas and alpacas in the United States and other countries. Whether the infection occurs in camelids in South America has previously been unknown. The current study documents a 15.8% infection rate among 76 Peruvian llamas, a 19.3% infection rate among Peruvian alpacas at one site, and a 9.26% infection rate in 108 Chilean alpacas from selected herds. All of the camelids tested appeared to be clinically healthy. No gender or species predilection was found. Only 1 positive camelid younger than 18 months was found. Infection is not associated with anemia, and the mean packed cell volume (PCV) in positive Peruvian camelids was slightly higher than the mean PCV in negative Peruvian camelids. In the Chilean alpacas, the positive alpacas had a slightly lower PCV than the negative alpacas, although the mean PCV was not in the anemic range in any of the groups.

Antibody responses in New World camelids with tuberculosis caused by Mycobacterium microti.

PubMed

Lyashchenko, K P; Greenwald, R; Esfandiari, J; Meylan, M; Burri, I Hengrave; Zanolari, P

2007-12-15

Antibody responses in New World camelids (NWC) infected with Mycobacterium microti were studied by two serological methods, multiantigen print immunoassay (MAPIA) and lateral-flow-based rapid test (RT). Serum samples were collected during 2004-2006 from 87 animals including 1 alpaca and 7 llamas with confirmed or suspected M. microti infection, 33 potentially exposed but clinically healthy animals from known infected herds, and 46 control NWC from herds where infection had not been previously diagnosed. The serological assays correctly identified infection status in 97% (MAPIA) or 87% (RT) cases. In three llamas with confirmed M. microti infection and one llama with gross pathology suggestive of disease, for which multiple serum samples collected over time were available, the antibody-based tests showed positive results 1-2 years prior to the onset of clinical signs or being found dead. In MAPIA, MPB83 protein was identified to be an immunodominant serological target antigen recognized in NWC infected with M. microti. With the limited number of animals tested in this study, the serological assays demonstrated the potential for convenient, rapid, and accurate diagnosis of M. microti infection in live llamas and alpacas.

Analysis of mitochondrial DNA in Bolivian llama, alpaca and vicuna populations: a contribution to the phylogeny of the South American camelids.

PubMed

Barreta, J; Gutiérrez-Gil, B; Iñiguez, V; Saavedra, V; Chiri, R; Latorre, E; Arranz, J J

2013-04-01

The objectives of this work were to assess the mtDNA diversity of Bolivian South American camelid (SAC) populations and to shed light on the evolutionary relationships between the Bolivian camelids and other populations of SACs. We have analysed two different mtDNA regions: the complete coding region of the MT-CYB gene and 513 bp of the D-loop region. The populations sampled included Bolivian llamas, alpacas and vicunas, and Chilean guanacos. High levels of genetic diversity were observed in the studied populations. In general, MT-CYB was more variable than D-loop. On a species level, the vicunas showed the lowest genetic variability, followed by the guanacos, alpacas and llamas. Phylogenetic analyses performed by including additional available mtDNA sequences from the studied species confirmed the existence of the two monophyletic clades previously described by other authors for guanacos (G) and vicunas (V). Significant levels of mtDNA hybridization were found in the domestic species. Our sequence analyses revealed significant sequence divergence within clade G, and some of the Bolivian llamas grouped with the majority of the southern guanacos. This finding supports the existence of more than the one llama domestication centre in South America previously suggested on the basis of archaeozoological evidence. Additionally, analysis of D-loop sequences revealed two new matrilineal lineages that are distinct from the previously reported G and V clades. The results presented here represent the first report on the population structure and genetic variability of Bolivian camelids and may help to elucidate the complex and dynamic domestication process of SAC populations. © 2012 The Authors, Animal Genetics © 2012 Stichting International Foundation for Animal Genetics.

LLAMA: normal star formation efficiencies of molecular gas in the centres of luminous Seyfert galaxies

NASA Astrophysics Data System (ADS)

Rosario, D. J.; Burtscher, L.; Davies, R. I.; Koss, M.; Ricci, C.; Lutz, D.; Riffel, R.; Alexander, D. M.; Genzel, R.; Hicks, E. H.; Lin, M.-Y.; Maciejewski, W.; Müller-Sánchez, F.; Orban de Xivry, G.; Riffel, R. A.; Schartmann, M.; Schawinski, K.; Schnorr-Müller, A.; Saintonge, A.; Shimizu, T.; Sternberg, A.; Storchi-Bergmann, T.; Sturm, E.; Tacconi, L.; Treister, E.; Veilleux, S.

2018-02-01

Using new Atacama Pathfinder Experiment and James Clerk Maxwell Telescope spectroscopy of the CO 2→1 line, we undertake a controlled study of cold molecular gas in moderately luminous (Lbol = 1043-44.5 erg s-1) active galactic nuclei (AGN) and inactive galaxies from the Luminous Local AGN with Matched Analogs (LLAMA) survey. We use spatially resolved infrared photometry of the LLAMA galaxies from 2MASS, the Wide-field Infrared Survey Explorer the Infrared Astronomical Satellite and the Herschel Space Observatory (Herschel), corrected for nuclear emission using multicomponent spectral energy distribution fits, to examine the dust-reprocessed star formation rates, molecular gas fractions and star formation efficiencies (SFEs) over their central 1-3 kpc. We find that the gas fractions and central SFEs of both active and inactive galaxies are similar when controlling for host stellar mass and morphology (Hubble type). The equivalent central molecular gas depletion times are consistent with the discs of normal spiral galaxies in the local Universe. Despite energetic arguments that the AGN in LLAMA should be capable of disrupting the observable cold molecular gas in their central environments, our results indicate that nuclear radiation only couples weakly with this phase. We find a mild preference for obscured AGN to contain higher amounts of central molecular gas, which suggests connection between AGN obscuration and the gaseous environment of the nucleus. Systems with depressed SFEs are not found among the LLAMA AGN. We speculate that the processes that sustain the collapse of molecular gas into dense pre-stellar cores may also be a prerequisite for the inflow of material on to AGN accretion discs.

Timing of mating and ovarian response in llamas (Lama glama) treated with pFSH.

PubMed

Ratto, M H; Gatica, R; Correa, J E

1997-08-01

The effect of the timing of mating on ovarian response in llamas was evaluated using 20 adult llamas weighing 90-120 kg which had been in oestrus for 5 days and were treated with 20 mg pFSH every 12 h for the following 5 days (total dose: 200 mg of FSH-NIH-P1). They were randomly allocated to Group A (N = 10) and mated immediately at the end of pFSH treatment or to Group B (n = 10) and mated 36 h after the end of pFSH treatment. Llamas of both groups were given hCG (750 iu, i.m.) immediately after mating. A second mating was allowed 12 h later. Ova and embryos were recovered by non-surgical uterine flushing 7 days after the first mating. Ovarian response was immediately evaluated afterwards via laparoscopy. The mean ovulation rate of 4.5 corpora lutea for Group A was significantly lower (P < 0.01) than the mean of 13.8 observed for Group B. The total ovarian response (number of corpora lutea + follicles > 10 mm) was also significantly higher (P < 0.01) in Group B than in Group A. Twenty-seven ova were recovered in each group, corresponding to 60% and 20% (P < 0.01) of the corpora lutea observed in Groups A and B, respectively; however, no significant difference (P > 0.05) in fertilisation rate was observed. The results show that pFSH induces superovulation in llamas treated during oestrus and that a 36-h interval between the end of FSH treatment and mating increases ovulation rate and the total ovarian response but does not affect the number of ova/embryos recovered.

Nitric oxide plays a role in the regulation of adrenal blood flow and adrenocorticomedullary functions in the llama fetus

PubMed Central

Riquelme, Raquel A; Sánchez, Gina; Liberona, Leonel; Sanhueza, Emilia M; Giussani, Dino A; Blanco, Carlos E; Hanson, Mark A; Llanos, Aníbal J

2002-01-01

The hypothesis that nitric oxide plays a key role in the regulation of adrenal blood flow and plasma concentrations of cortisol and catecholamines under basal and hypoxaemic conditions in the llama fetus was tested. At 0.6-0.8 of gestation, 11 llama fetuses were surgically prepared for long-term recording under anaesthesia with vascular and amniotic catheters. Following recovery all fetuses underwent an experimental protocol based on 1 h of normoxaemia, 1 h of hypoxaemia and 1 h of recovery. In nine fetuses, the protocol occurred during fetal i.v. infusion with saline and in five fetuses during fetal i.v. treatment with the nitric oxide synthase inhibitor l-NAME. Adrenal blood flow was determined by the radiolabelled microsphere method during each of the experimental periods during saline infusion and treatment with l-NAME. Treatment with l-NAME during normoxaemia led to a marked fall in adrenal blood flow and a pronounced increase in plasma catecholamine concentrations, but it did not affect plasma ACTH or cortisol levels. In saline-infused fetuses, acute hypoxaemia elicited an increase in adrenal blood flow and in plasma ACTH, cortisol, adrenaline and noradrenaline concentrations. Treatment with l-NAME did not affect the increase in fetal plasma ACTH, but prevented the increments in adrenal blood flow and in plasma cortisol and adrenaline concentrations during hypoxaemia in the llama fetus. In contrast, l-NAME further enhanced the increase in fetal plasma noradrenaline. These data support the hypothesis that nitric oxide has important roles in the regulation of adrenal blood flow and adrenal corticomedullary functions during normoxaemia and hypoxaemia functions in the late gestation llama fetus. PMID:12356897

Pulmonary Arterial Lesions in New World Camelids in Association With Dicrocoelium dendriticum and Fasciola hepatica Infection.

PubMed

Hilbe, M; Robert, N; Pospischil, A; Gerspach, C

2015-11-01

In Switzerland, dicrocoeliasis is regarded as the most significant parasitic infection of llamas and alpacas. Fasciola hepatica infestation is also a problem but less common. The aim of the present retrospective study was to evaluate the lungs of New World camelids (NWCs) for evidence of arterial hypertension in association with liver changes due to liver fluke infestation. The lungs of 20 llamas and 20 alpacas with liver fluke infestation were histologically evaluated. The hematoxylin and eosin and van Gieson (VG)-elastica stains as well as immunohistology for the expression of α-smooth muscle actin (α-SMA) were used to visualize the structures of arterial walls. Parasitology of fecal matter (11 llamas and 17 alpacas) confirmed that most of these animals were infested with both Dicrocoelium dendriticum and other gastrointestinal parasites. In most cases (10/12 llamas, 4/6 alpacas), liver enzyme activity in serum was elevated. Histologically, arteries in the lungs of 9 of 20 llamas (45%) and 3 of 20 alpacas (15%) showed severe intimal and adventitial and slight to moderate medial thickening, which was confirmed with α-SMA and VG-elastica staining. All animals exhibited typical liver changes, such as fibrosis and biliary hyperplasia, in association with the presence of liver flukes. This study shows that liver flukes can induce proliferative changes in lung arteries in NWCs that resemble those seen with pulmonary arterial hypertension due to liver parasites in humans. However, the degree of liver fluke infestation was not correlated with the extent of liver damage, or with the amount of thoracic or abdominal effusion or pulmonary arterial changes. © The Author(s) 2015.

Microbial ecology involved in the ripening of naturally fermented llama meat sausages. A focus on lactobacilli diversity.

PubMed

Fontana, Cecilia; Bassi, Daniela; López, Constanza; Pisacane, Vincenza; Otero, Maria Claudia; Puglisi, Edoardo; Rebecchi, Annalisa; Cocconcelli, Pier Sandro; Vignolo, Graciela

2016-11-07

Llama represents for the Andean regions a valid alternative to bovine and pork meat and thanks to the high proteins and low fat content; it can constitute a good product for the novel food market. In this study, culture-dependent and independent methods were applied to investigate the microbial ecology of naturally fermented llama sausages produced in Northwest Argentina. Two different production technologies of llama sausage were investigated: a pilot-plant scale (P) and an artisanal one (A). Results obtained by High-Throughput Sequencing (HTS) of 16S rRNA amplicons showed that the production technologies influenced the development of microbial communities with a different composition throughout the entire fermentation process. Both sequencing and microbiological counts demonstrated that Lactic Acid Bacteria (LAB) contributed largely to the dominant microbiota. When a total of 230 isolates were approached by RAPD-PCR, presumptive LAB strains from P production exhibited an initial variability in RAPD fingerprints switching to a single profile at the final of ripening, while A production revealed a more heterogeneous RAPD pattern during the whole fermentation process. The constant presence of Lactobacillus sakei along the fermentation in both productions was revealed by HTS and confirmed by species-specific PCR from isolated strains. The technological characterization of Lb. sakei isolates evidenced their ability to grow at 15°C, pH4.5 and 5% NaCl (95%). Most strains hydrolyzed myofibrillar and sarcoplasmic proteins. Bacteriocins encoding genes and antimicrobial resistance were found in 35% and 42.5% of the strains, respectively. An appropriate choice of a combination of autochthonous strains in a starter formulation is fundamental to improve and standardize llama sausages safety and quality. Copyright © 2016 Elsevier B.V. All rights reserved.

The Adventures of Artemis and the Llama: A Case for Imaginary Histories in Art Education

ERIC Educational Resources Information Center

Vallance, Elizabeth

2004-01-01

Artemis is a late Hellenistic Greek marble sculpture of the huntress, running in a flowing garment, now lacking arms, legs, and head, and about three-quarters life-sized. The llama is a remarkable hollow male figure of smooth thin gold, and about two inches tall, and was made by the Inca before the Spanish conquest in 1532. This narrative is just…

Coccidioidomycosis in the llama: case report and epidemiologic survey.

PubMed

Muir, S; Pappagianis, D

1982-12-01

An 8-year-old nongravid female llama with a 1-month history of progressive posterior paresis was referred because of suspected degenerative myelopathy secondary to copper deficiency or plant poisoning. Neurologic examination revealed loss of conscious proprioception and slightly depressed bilateral patellar reflexes. Electromyographic examination of hindlimb flexors and extensors did not elicit evidence of lower motor neuron disease. Possible fragmentation and mottling of the 10th thoracic vertebral body were noted radiographically. Results of a lumbar CSF tap, complete blood count, and fecal flotation were not diagnostic. In the face of poor prognosis, the llama was euthanatized. Postmortem and histologic evaluation revealed, in addition to disseminated visceral granulomas, an extradural pyogranulomatous mass compressing the cord laterally at the level of T-10. Large numbers of Coccidioides immitis were dispersed throughout the granulomas. Complement fixing antibody tests in 11 other herd members showed evidence of C immitis infection in three.

Adaptive functional specialisation of architectural design and fibre type characteristics in agonist shoulder flexor muscles of the llama, Lama glama.

PubMed

Graziotti, Guillermo H; Chamizo, Verónica E; Ríos, Clara; Acevedo, Luz M; Rodríguez-Menéndez, J M; Victorica, C; Rivero, José-Luis L

2012-08-01

Like other camelids, llamas (Lama glama) have the natural ability to pace (moving ipsilateral limbs in near synchronicity). But unlike the Old World camelids (bactrian and dromedary camels), they are well adapted for pacing at slower or moderate speeds in high-altitude habitats, having been described as good climbers and used as pack animals for centuries. In order to gain insight into skeletal muscle design and to ascertain its relationship with the llama's characteristic locomotor behaviour, this study examined the correspondence between architecture and fibre types in two agonist muscles involved in shoulder flexion (M. teres major - TM and M. deltoideus, pars scapularis - DS and pars acromialis - DA). Architectural properties were found to be correlated with fibre-type characteristics both in DS (long fibres, low pinnation angle, fast-glycolytic fibre phenotype with abundant IIB fibres, small fibre size, reduced number of capillaries per fibre and low oxidative capacity) and in DA (short fibres, high pinnation angle, slow-oxidative fibre phenotype with numerous type I fibres, very sparse IIB fibres, and larger fibre size, abundant capillaries and high oxidative capacity). This correlation suggests a clear division of labour within the M. deltoideus of the llama, DS being involved in rapid flexion of the shoulder joint during the swing phase of the gait, and DA in joint stabilisation during the stance phase. However, the architectural design of the TM muscle (longer fibres and lower fibre pinnation angle) was not strictly matched with its fibre-type characteristics (very similar to those of the postural DA muscle). This unusual design suggests a dual function of the TM muscle both in active flexion of the shoulder and in passive support of the limb during the stance phase, pulling the forelimb to the trunk. This functional specialisation seems to be well suited to a quadruped species that needs to increase ipsilateral stability of the limb during the support phase of the pacing gait. Compared with other species, llama skeletal muscles are well suited for greater force generation combined with higher fatigue resistance during exercise. These characteristics are interpreted as being of high adaptive value, given the llama's habitat and its use as a pack animal. © 2012 The Authors. Journal of Anatomy © 2012 Anatomical Society.

Evaluation of insulin secretion and action in New World camelids.

PubMed

Firshman, Anna M; Cebra, Christopher K; Schanbacher, Barbara J; Seaquist, Elizabeth R

2013-01-01

To measure and compare insulin secretion and sensitivity in healthy alpacas and llamas via glucose clamping techniques. 8 llamas and 8 alpacas. Hyperinsulinemic euglycemic clamping (HEC) and hyperglycemic clamping (HGC) were performed on each camelid in a crossover design with a minimum 48-hour washout period between clamping procedures. The HEC technique was performed to measure insulin sensitivity. Insulin was infused IV at 6 mU/min/kg for 4 hours, and an IV infusion of glucose was adjusted to maintain blood glucose concentration at 150 mg/dL. Concentrations of blood glucose and plasma insulin were determined throughout. The HGC technique was performed to assess insulin secretion in response to exogenous glucose infusion. An IV infusion of glucose was administered to maintain blood glucose concentration at 320 mg/dL for 3 hours, and concentrations of blood glucose and plasma insulin were determined throughout. Alpacas and llamas were not significantly different with respect to whole-body insulin sensitivity during HEC or in pancreatic β-cell response during HGC. Alpacas and llamas had markedly lower insulin sensitivity during HEC and markedly lower pancreatic β-cell response during HGC, in comparison with many other species. New World camelids had lower glucose-induced insulin secretion and marked insulin resistance in comparison with other species. This likely contributes to the disorders of fat and glucose metabolism that are common to camelids.

Toxoplasma gondii and Neospora caninum seroprevalences in domestic South American camelids of the Peruvian Andes.

PubMed

Chávez-Velásquez, Amanda; Aguado-Martínez, Adriana; Ortega-Mora, Luis M; Casas-Astos, Eva; Serrano-Martínez, Enrique; Casas-Velásquez, Gina; Ruiz-Santa-Quiteria, Jose A; Alvarez-García, Gema

2014-10-01

The objective of this study was to investigate the presence of Toxoplasma gondii- and Neospora caninum-specific antibodies in domestic South American camelids (SAC) (llamas and alpacas) from the Peruvian Andes through a cross-sectional study. A wide panel of serum samples collected from 1,845 llamas and 2,874 alpacas from the two main SAC production areas of Peru was selected. Immunofluorescence antibody technique was employed to detect and titrate specific anti-T. gondii and anti-N. caninum immunoglobulins G in serum samples. The association between T. gondii and N. caninum seroprevalence and the geographical origin (Central and South Peruvian Andes) was evaluated. Anti-T. gondii antibodies were found in 460 (24.9 %) llamas and 706 (24.6 %) alpacas, whereas anti-N. caninum antibodies were detected in 153 (8.3 %) llamas and 425 (14.8 %) alpacas. Toxoplasma gondii infection was strongly associated with the South Peruvian Andes where moderate climate conditions, larger human population, compared to the Central region, and the presence of wildlife definitive hosts could favor horizontal transmission to SAC. In contrast, N. caninum infection was not associated with the geographical region. These results indicate that T. gondii and N. caninum infections are highly and moderately widespread, respectively, in both species of domestic SAC studied in the sampled areas and appropriate control measures should be undertaken to reduce the prevalence of both parasitic infections.

Direct colloid osmometry in healthy New World camelids.

PubMed

Quesada, Rolando J; Gorman, Maria Elena; Cebra, Christopher K; Verdugo, Claudio; Mosley, Craig A

2011-06-01

Direct colloid osmometry provides an objective assessment of the oncotic effects of crystalloid or colloidal fluid therapy, which is especially useful in monitoring fluid therapy of critically ill camelids due to their tendency toward nonspecific hypoproteinemia with increased risk of developing edema and ascites. The aims of this study were to measure colloid osmotic pressure (COP) of alpacas and llamas, determine its correlation with concentrations of total protein (TP) and total solids (TS), as well as both albumin (A) and globulin (G) concentrations in the same model (A+G), and evaluate the effects of sample type and storage conditions on COP. Blood was collected from clinically healthy alpacas (n=23) and llamas (n=22) into heparin tubes. COP of fresh whole blood (COP(FB) ) and plasma (COP(FP) ) was determined using a membrane osmometer. For 20 alpacas, COP of refrigerated whole blood (COP(RB) ) and frozen plasma (COP(FrP) ) was also measured. Correlations between COP(FB) and TS, TP, and A+G concentrations were assessed by simple and multiple regression analysis to model potential predictors. Median COP(FB) from alpacas (24.6 mmHg, range 19.3-28.1) was not significantly different from that of llamas (25.3 mmHg, range 22.5-33.7). Sample type or storage conditions did not affect COP. Measured COP had a strong positive linear correlation with TS, TP, and A+G concentrations in alpacas (r(2) =.7, .74, and .88, respectively). In llamas, COP correlated best with TS concentration (r(2) =.59), whereas correlation with TP and A+G concentrations was poor (r(2) =.19 and .25, respectively). COP can be measured using heparinized whole blood or plasma, either fresh or stored. Direct measurement is recommended whenever quantitative knowledge of COP is required in clinical or research setting. Further studies are needed to verify if the poor association of COP with TP found in this study can be generalized to llamas. ©2011 American Society for Veterinary Clinical Pathology.

Nutritional requirements and assessing nutritional status in camelids.

PubMed

Van Saun, Robert J

2009-07-01

It has been nearly 30 years since the first imported llamas and alpacas have been commercially raised in the United States. Nutritional requirements for these animals have not been well understood and most feeding practices were based on extrapolated and experiential information. Only recently has a National Research Council committee reviewed the available published information relative to nutrient requirements of llamas and alpacas. This article summarizes current nutrient requirement recommendations and provides some practical feeding recommendations and methods to assess nutritional status.

Third compartment ulcers in the llama.

PubMed

Smith, B B; Pearson, E G; Timm, K I

1994-07-01

The diagnosis of third compartment ulcers in the llama and alpaca is largely one of exclusion. Clinical signs may include mild to severe colic, inappetence, decreased fecal output, bruxism, and depression. Abdominocentesis results are usually unremarkable if C3 perforation has not occurred but reflective of a generalized peritonitis if full thickness ulceration has occurred. The H-2 receptor antagonists cimetidine and ranitidine do not suppress C3 acid production for a significant period of time and are of questionable efficacy in the management of C3 ulcers.

Estimating total body water content in suckling and lactating llamas (Lama glama) by isotope dilution.

PubMed

Riek, Alexander; Gerken, Martina

2010-08-01

Total body water (TBW) in 17 suckling and six lactating llamas was estimated from isotope dilution at three different post natum and lactation stages using both (18)O and deuterium oxide (D(2)O). In total, 69 TBW measurements were undertaken. While TBW in lactating dams, expressed in kilogram, remained stable during the three measurement periods (91.8 +/- 15.0 kg), the body water fraction (TBW expressed in percent of body mass) increased slightly (P = 0.042) from 62.9% to 65.8%. In contrast, TBW (kilogram) in suckling llamas increased significantly (P < 0.001) with age and decreased slightly when expressed as a percentage of body mass (P = 0.016). Relating TBW to body mass across all animals yielded a highly significant regression equation (TBW in kilogram = 2.633 + 0.623 body mass in kilogram, P < 0.001, n = 69) explaining 99.5% of the variation. The water fraction instead decreased in a curve linear fashion with increasing body mass (TBW in percent of body mass = 88.23 body mass in kilogram(-0.064), P < 0.001, R (2) = 0.460). The present results on TBW can serve as reference values for suckling and lactating llamas, e.g., for the evaluation of fluid losses during disease. Additionally, the established regression equations can be used to predict TBW from body mass, providing that the body masses fall inside the range of masses used to derive the equations.

Hepatic lipidosis in llamas and alpacas: 31 cases (1991-1997).

PubMed

Tornquist, S J; Van Saun, R J; Smith, B B; Cebra, C K; Snyder, S P

1999-05-01

To identify factors associated with hepatic lipidosis (HL) in llamas and alpacas. Retrospective case series. 30 llamas and 1 alpaca. Medical records were searched to identify llamas or alpacas in which a histologic diagnosis of HL was made. Information was retrieved on signalment, history, clinical and laboratory findings, and results of necropsy or examination of biopsy specimens. Data were analyzed using descriptive statistics and chi 2 analyses. Females were affected more often than males; however, the sex distribution was not different from that of the camelid population in the diagnostic laboratory's database. Fifty-four percent of the females were pregnant, and 46% were lactating. Most affected camelids were 6 to 10 years old. Anorexia and recent weight loss were common (51.6% of camelids). An infective agent was found in only one ilama, and toxins and mineral deficiencies were not identified. The most common abnormalities on serum biochemical analysis were a high concentration of bile acids, high activities of gamma-glutamyltransferase (GGT) and aspartate aminotransferase (AST), and hypoproteinemia. Concentrations of nonesterified fatty acids (NEFA) and beta-hydroxybutyrate (beta-HB) were high in those camelids in which these compounds were assayed. Twenty-nine camelids did not survive. Sick camelids should be considered at risk for developing HL, especially those with anorexia or the metabolic demands of pregnancy and lactation. Other stresses also appear to contribute. High concentrations of NEFA, beta-HB, and bile acids; high activities of GGT and AST; and hypoproteinemia may indicate that HL has developed.

Characterization of Fusobacterium necrophorum isolated from llama and alpaca.

PubMed

Kumar, Amit; Anderson, David; Amachawadi, Raghavendra G; Nagaraja, Tiruvoor G; Narayanan, Sanjeev K

2013-07-01

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, pharynx, interdigital spaces, foot pad, larynx, mandible, or maxillary bones. The objective of the current study was to characterize the presumptive Fusobacterium isolates from a variety of necrotic infections in llama (Lama glama) and alpaca (Vicugna pacos) and determine whether the strains possess leukotoxin activities. A total of 7 isolates from alpaca and 2 isolates from llama were characterized. Based on growth characteristics in broth culture, and biochemical and polymerase chain reaction analyses, all 9 isolates belonged to subsp. necrophorum and possessed the putative hemagglutinin gene. Western blot analysis with antileukotoxin antibodies raised in rabbit showed the presence of leukotoxin protein in the culture supernatant of all isolates. Furthermore, flow cytometry of the culture supernatants demonstrated cytotoxicity to bovine and alpaca polymorphonuclear leukocytes (PMNs). The extent of cytotoxicity to either alpaca or bovine PMNs differed among camelid strains. The cytotoxicity of many of the camelid strains was higher (P < 0.05) toward alpaca PMNs compared to bovine PMNs. Fusobacterium necrophorum isolates from llama and alpaca are similar to bovine isolates, and leukotoxin may be a major virulence factor.

Contrast-enhanced computed tomography of the gastrointestinal tract in clinically normal alpacas and llamas.

PubMed

Stieger-Vanegas, Susanne M; Cebra, Christopher K

2013-01-15

To assess the feasibility and usefulness of CT enterography to evaluate the gastrointestinal tract in clinically normal llamas and alpacas. Prospective observational study. 7 clinically normal alpacas and 8 clinically normal llamas. The imaging protocol included orogastric administration of iodinated contrast material mixed with water. Three hours later, helical CT scanning was performed of the entire abdomen with transverse and multiplanar sagittal and dorsal projections before and after IV iodinated contrast agent injection. Both oral and IV contrast agents were well tolerated, and no adverse reactions were observed. Transverse images depicted the gastrointestinal tract and pancreas in the short axis; however, dorsal and sagittal projections aided in localizing and differentiating the various gastrointestinal segments, including the pancreas. In all camelids, the wall of the gastrointestinal tract was well differentiated. In all but 2 camelids, all gastrointestinal segments were well visualized and differentiated. In those 2 animals, the cecum was difficult to identify. Good distention of the small intestine was achieved by use of the oral contrast agent. The dorsal projections were useful to identify the pancreas in its entire length. The present study supplied new information about gastrointestinal wall thickness, intestinal diameter, and location of the pancreas and ileocecocolic junction in alpacas and llamas. Multiplanar contrast-enhanced CT was useful to reveal the various segments of the gastrointestinal tract, pancreas, and abdominal lymph nodes. The shorter time delay before imaging, compared with the delay with conventional barium studies, makes this technique complementary or superior to conventional radiographic or ultrasonographic studies for evaluation of the gastrointestinal tract.

Nutritional support for treatment of hepatic lipidosis in a llama.

PubMed

Van Saun, R J; Callihan, B R; Tornquist, S J

2000-11-15

A 3-year-old female llama that was 3 months into her first lactation and 10 weeks pregnant was evaluated for anorexia of 24 hours' duration. On physical examination, the llama was in lateral recumbency, bradycardic, tachypneic, and hyperthermic. Palpation per rectum confirmed the presence of a possible dry fecal mass in the spiral colon. A tissue biopsy specimen of the liver was obtained, and histologic examination revealed moderate diffuse lipid accumulation within the hepatocytes. Lactated Ringer's solution was administered for rehydration, and partial parenteral nutrition was then initiated. Hepatic lipidosis is a disease characterized by abnormal accumulation of lipid in the liver and is associated with high mortality in camelids. Anorexia associated with hepatic lipidosis promotes further lipid mobilization and fatty infiltration of the liver. Partial parenteral nutrition with enteral supplementation may be used to maintain adequate energy intake and minimize further lipid mobilization. The distinctive metabolism of camelids may require higher amino acid supplementation relative to nonprotein calories in parenteral solutions than those traditionally provided to other species. Treatment with insulin may be effective

Comparison of the effect of natural mating, LH, and GnRH on interval to ovulation and luteal function in llamas.

PubMed

Ratto, Marcelo; Huanca, Wilfredo; Singh, Jaswant; Adams, Gregg P

2006-02-01

Gonadotropins and GnRH have been used to electively induce ovulation in llamas and alpacas, but critical evaluation of the natural interval to ovulation after mating has not been performed nor has a direct comparison of the effects of natural mating versus hormone treatments on this interval and subsequent luteal development. The objectives of this study were to compare the effects of hormonal treatments and natural mating on ovulation induction, interval to ovulation, and luteal development in llamas. The ovaries of llamas were examined by transrectal ultrasonography once daily. Llamas with a large follicle were assigned randomly to be: (1) mated with an intact male (mated; n=10); (2) given 5 mg of LH im (LH; n=11); or (3) 50 microg of GnRH im (GnRH; n=10). Ultrasound examinations were performed every 4h from treatment (day 0) to ovulation and thereafter once daily for 15 consecutive days to monitor CL growth and regression (n=5 per group). Plasma progesterone concentrations were measured at days 0, 3, 6, 9, and 12 after treatment to evaluate CL function. The size of the largest preovulatory follicle at the time of treatment did not differ among groups (11+/-0.6, 10.5+/-0.8, 11.8+/-0.9 mm, for mated, LH, and GnRH groups, respectively; P=0.6). No differences were detected among groups (mated, LH, and GnRH) in ovulation rate (80%, 91%, 80%, respectively; P=0.6), or interval from treatment to ovulation (30.0+/-0.5, 29.3+/-0.6, 29.3+/-0.7h, respectively; P=0.9). Similarly, no differences were detected among groups (mated, LH, and GnRH) in maximum CL diameter (14.2+/-0.3, 13.2+/-0.5, and 13.0+/-0.7 mm, respectively; P=0.5), the day of maximum CL diameter (7.6+/-0.2, 7.6+/-0.2, and 7.4+/-0.4 mm, respectively; P=0.6), or the day on which the CL began to regress (12.3+/-0.3 [non-pregnant, n=3], 11.8+/-0.6, 12.2+/-0.4, respectively; P=0.4). The diameter of the CL and plasma progesterone concentrations changed over days (P<0.0001) but the profiles did not differ among groups. In summary, ovulation rate, interval to ovulation, and luteal development were similar among llamas that were mated naturally or treated with LH or GnRH. We conclude that both hormonal preparations are equally reliable for inducing ovulation and suitable for synchronization for artificial insemination or embryo transfer program.

Assessment of clinical application of pulse oximetry probes in llamas and alpacas.

PubMed

Grubb, Tamara L; Anderson, David E

2017-08-01

The placement and accuracy of pulse oximeter probes can vary markedly among species. For our study, we aimed to assess the accuracy of pulse oximetry and to determine the most clinically useful sites for probe placement in llamas and alpacas. The objectives included an analysis of pulse oximetry probes for accurate assessment of llamas and alpacas and to determine the best placement of the probes to achieve accurate readings. For study 1, saturation of haemoglobin with oxygen was measured in 184 arterial blood gas samples (SaO 2 ) using a co-oximeter and compared to saturation of haemoglobin with oxygen simultaneously measured using a pulse oximeter (S p O 2 ). The bias and precision for the SpO 2 -S a O 2 difference was calculated and plotted on a Bland-Altman plot. For study 2, S p O 2 data was collected 624 times from a variety of sites [tongue (T), nasal septum (NS), lip (L), vulva (V), prepuce (P), ear (E), and scrotum (S)] and recorded based upon a percentage of successful readings. Results for study 1 revealed that S p O 2 was consistently 0 to -6% points different than S a O 2 . The bias and precision of the S p O 2 -S a O 2 difference was -2.6 ± 1.7%. Results for study 2 uncovered that 540 recordings were successful readings and were obtained from the tongue and nasal septum with 97% accuracy, the lip 80%, vulva 62%, prepuce 59%, ear and scrotum < 50%. We concluded that pulse oximetry probes provide reliable estimates of arterial haemoglobin oxygen saturation in llamas and alpacas and is most accurately read when placed on the nasal septum or tongue.

Y-chromosome and mtDNA variation confirms independent domestications and directional hybridization in South American camelids.

PubMed

Marín, J C; Romero, K; Rivera, R; Johnson, W E; González, B A

2017-10-01

Investigations of genetic diversity and domestication in South American camelids (SAC) have relied on autosomal microsatellite and maternally-inherited mitochondrial data. We present the first integrated analysis of domestic and wild SAC combining male and female sex-specific markers (male specific Y-chromosome and female-specific mtDNA sequence variation) to assess: (i) hypotheses about the origin of domestic camelids, (ii) directionality of introgression among domestic and/or wild taxa as evidence of hybridization and (iii) currently recognized subspecies patterns. Three male-specific Y-chromosome markers and control region sequences of mitochondrial DNA are studied here. Although no sequence variation was found in SRY and ZFY, there were seven variable sites in DBY generating five haplotypes on the Y-chromosome. The haplotype network showed clear separation between haplogroups of guanaco-llama and vicuña-alpaca, indicating two genetically distinct patrilineages with near absence of shared haplotypes between guanacos and vicuñas. Although we document some examples of directional hybridization, the patterns strongly support the hypothesis that llama (Lama glama) is derived from guanaco (Lama guanicoe) and the alpaca (Vicugna pacos) from vicuña (Vicugna vicugna). Within male guanacos we identified a haplogroup formed by three haplotypes with different geographical distributions, the northernmost of which (Peru and northern Chile) was also observed in llamas, supporting the commonly held hypothesis that llamas were domesticated from the northernmost populations of guanacos (L. g. cacilensis). Southern guanacos shared the other two haplotypes. A second haplogroup, consisting of two haplotypes, was mostly present in vicuñas and alpacas. However, Y-chromosome variation did not distinguish the two subspecies of vicuñas. © 2017 Stichting International Foundation for Animal Genetics.

Cutaneous angiomatosis in a llama (Lama glama).

PubMed

Luppi, M M; Malta, M C C; Ocarino, N M; França, S A; Serakides, R

2010-01-01

Cutaneous angiomatosis was diagnosed in an adult female llama (Lama glama). Lesions were raised or plaque-like, erythematous, firm to soft in consistency and were observed on the face and skin of the axillary, abdominal, perineal and inguinal regions. The lesions were not painful or pruritic. Microscopical examination revealed an irregular parakeratotic lamellar hyperkeratosis associated with diffuse proliferation of arterioles and venules in the superficial dermis. Immunohistochemical analysis determined that the cells forming these vessels and perivascular cells expressed factor VIII-related antigen, vascular endothelial growth factor (VEGF), CD31 and smooth-muscle alpha-actin. These studies confirmed the diagnosis of cutaneous angiomatosis. Copyright 2009 Elsevier Ltd. All rights reserved.

Llama oviductal sperm reservoirs: involvement of bulbourethral glands.

PubMed

Apichela, S A; Argañaraz, M E; Giuliano, S; Zampini, R; Carretero, I; Miragaya, M; Miceli, D C

2014-04-01

The aim of this study was to elucidate the role of llama seminal plasma in the formation of oviductal sperm reservoirs. Female llamas with follicles in the mature phase were mated with a bulbourethral glands-removed male. Females mated with nonbulbourethral glands-removed males were used as control. Oviducts were obtained by surgery 24 h after mating. The uterotubal junction and isthmus were examined by scanning electron microscopy, and mucopolysaccharides were identified by Alcian blue staining. To know the proteins probably involved in sperm reservoir formation, SDS-PAGE of seminal plasma (8% and 18% resolving gel) was made. Spermatozoa only adhered to the oviductal mucosa surface of uterotubal junction of females mated with nonbulbourethral glands-removed males confirming that seminal plasma and, in particular, bulbourethral secretions are related with the oviductal sperm reservoir formation. Histological sections showed sperm in the lumen, immersed in substance, positive for acid mucopolysaccharides. Alcian blue staining of seminal plasma proteins SDS-PAGE showed a band of high molecular weight containing mucopolysaccharides, only present in nonbulbourethral glands-removed males. Bulbourethral glands would secrete at least eight different proteins that most likely participate in the process of sperm storage in the oviduct. © 2013 Blackwell Verlag GmbH.

Serology and longevity of immunity against Echinococcus granulosus in sheep and llama induced by an oil-based EG95 vaccine.

PubMed

Poggio, T V; Jensen, O; Mossello, M; Iriarte, J; Avila, H G; Gertiser, M L; Serafino, J J; Romero, S; Echenique, M A; Dominguez, D E; Barrios, J R; Heath, D

2016-08-01

An oil-based formulation of the EG95 vaccine to protect grazing animals against infection with Echinococcus granulosus was formulated in Argentina. The efficacy of the vaccine was monitored by serology in sheep and llama (Lama glama) and was compared to the serology in sheep previously published using a QuilA-adjuvanted vaccine. Long-term efficacy was also tested in sheep by challenging with E. granulosus eggs of the G1 strain 4 years after the beginning of the trial. The serological results for both sheep and llama were similar to those described previously, except that there was a more rapid response after the first vaccination. A third vaccination given after 1 year resulted in a transient boost in serology that lasted for about 12 months, which was similar to results previously described. Sheep challenged after 4 years with three vaccinations presented 84·2% reduction of live cysts counts compared with control group, and after a fourth vaccination prior to challenge, this reduction was 94·7%. The oil-based vaccine appeared to be bio-equivalent to the QuilA vaccine. © 2016 John Wiley & Sons Ltd.

Evaluation of insulin resistance in two kinds of South American camelids: llamas and alpacas.

PubMed

Araya, A V; Atwater, I; Navia, M A; Jeffs, S

2000-10-01

Insulin resistance was evaluated in South American camelids, llamas and alpacas, by use of the minimal model test and the insulin tolerance test. Animals were catheterized for long-term studies and tamed to minimize stress during evaluation. Results indicated a low insulin sensitivity index (SI) = 0 to 0.97, median = 0.39 x 10(-4) min/uIU x ml, about a fifth the value in other mammals and humans. The KITT was between 1.43 and 3.19 %/min, also significantly lower than that reported for humans. Glycosylated hemoglobin concentration was 6%, and HbAlc concentration was 5.5%; red blood cell lifetime, as measured by use of the 51Cr method, was 120 days, similar to the value in humans. We concluded that llamas and alpacas have naturally higher blood glucose concentration than do humans and other mammals during the glucose tolerance test. Using the same mathematical tools to evaluate glucose metabolism as those used in people, South American camelids appear to be resistant to insulin. Thus, the South American camelid may be a useful new animal model for the study of sugar metabolism and various facets of diabetes mellitus, especially protection from the deleterious effects of glycosylation.

In Vitro Neutralisation of Rotavirus Infection by Two Broadly Specific Recombinant Monovalent Llama-Derived Antibody Fragments

PubMed Central

Aladin, Farah; Einerhand, Alexandra W. C.; Bouma, Janneke; Bezemer, Sandra; Hermans, Pim; Wolvers, Danielle; Bellamy, Kate; Frenken, Leon G. J.; Gray, Jim; Iturriza-Gómara, Miren

2012-01-01

Rotavirus is the main cause of viral gastroenteritis in young children. Therefore, the development of inexpensive antiviral products for the prevention and/or treatment of rotavirus disease remains a priority. Previously we have shown that a recombinant monovalent antibody fragment (referred to as Anti-Rotavirus Proteins or ARP1) derived from a heavy chain antibody of a llama immunised with rotavirus was able to neutralise rotavirus infection in a mouse model system. In the present work we investigated the specificity and neutralising activity of two llama antibody fragments, ARP1 and ARP3, against 13 cell culture adapted rotavirus strains of diverse genotypes. In addition, immunocapture electron microscopy (IEM) was performed to determine binding of ARP1 to clinical isolates and cell culture adapted strains. ARP1 and ARP3 were able to neutralise a broad variety of rotavirus serotypes/genotypes in vitro, and in addition, IEM showed specific binding to a variety of cell adapted strains as well as strains from clinical specimens. These results indicated that these molecules could potentially be used as immunoprophylactic and/or immunotherapeutic products for the prevention and/or treatment of infection of a broad range of clinically relevant rotavirus strains. PMID:22403728

Llama medicine. Nutrition.

PubMed

Johnson, L W

1989-03-01

The NWC are capable of assimilating a much lower plane of nutrition than we are accustomed to offer conventional domestic herbivores. Their unique gastric anatomy and physiology no doubt contribute to this apparent superiority, which is most evident when compared with sheep under lesser planes of nutrition. Llamas tend to be "cafeteria style" eaters, with some preference being given to taller, coarser forages. Absolute maintenance requirements for caloric, protein, fiber, and mineral content of NWC rations are yet to be finalized using North American feedstuffs, but some guidelines are offered. Proper assessment of body condition, acknowledgment of any nutritional problems, ration analysis, and assessment of management procedures likely will keep NWC nutrition on an even plane here in their new-found environment.

Fish and mammals in the economy of an ancient Peruvian kingdom

PubMed Central

Marcus, Joyce; Sommer, Jeffrey D.; Glew, Christopher P.

1999-01-01

Fish and mammal bones from the coastal site of Cerro Azul, Peru shed light on economic specialization just before the Inca conquest of A.D. 1470. The site devoted itself to procuring anchovies and sardines in quantity for shipment to agricultural communities. These small fish were dried, stored, and eventually transported inland via caravans of pack llamas. Cerro Azul itself did not raise llamas but obtained charqui (or dried meat) as well as occasional whole adult animals from the caravans. Guinea pigs were locally raised. Some 20 species of larger fish were caught by using nets; the more prestigious varieties of these show up mainly in residential compounds occupied by elite families. PMID:10339628

Quantitative determination of H2-utilizing acetogenic and sulfate-reducing bacteria and methanogenic archaea from digestive tract of different mammals.

PubMed

Morvan, B; Bonnemoy, F; Fonty, G; Gouet, P

1996-03-01

Total number of bacteria, cellulolytic bacteria, and H2-utilizing microbial populations (methanogenic archaea, acetogenic and sulfate-reducing bacteria) were enumerated in fresh rumen samples from sheep, cattle, buffaloes, deer, llamas, and caecal samples from horses. Methanogens and sulfate reducers were found in all samples, whereas acetogenes were not detected in some samples of each animal. Archaea methanogens were the largest H2-utilizing populations in all animals, and a correlation was observed between the numbers of methanogens and those of cellulolytic microorganisms. Higher counts of acetogens were found in horses and llamas (1 x 10(4) and 4 x 10(4) cells ml-1 respectively).

Using llama derived single domain antibodies to target botulinum neurotoxins

NASA Astrophysics Data System (ADS)

Swain, Marla D.; Anderson, George P.; Bernstein, Rachael D.; Liu, Jinny L.; Goldman, Ellen R.

2010-04-01

Llama serum contains both conventional IgG as well as unique forms of antibody that contain only heavy chains where antigen binding is mediated through a single variable domain. These variable domains can be expressed recombinantly and are referred to as single domain antibodies (sdAb). SdAb are among the smallest known naturally derived antigen binding fragments, possess good solubility, thermal stability, and can refold after heat and chemical denaturation. Llamas were immunized with either BoNT A or B toxoid and phage display libraries prepared. Single domain antibodies (sdAb) that were able to detect botulinum neurotoxin (BoNT) serotypes A and B were selected from their respective libraries. Here, the binders obtained by panning the BoNT B library on either BoNT B toxoid or BoNT B complex toxoid coated plates or BoNT B toxin coupled microspheres are described. Using these panning methods, we selected for binders that showed specificity for BoNT B. Phage displayed binders were screened, moved to a protein expression vector and soluble sdAb was produced. Using a Luminex flow cytometer binders were evaluated in direct binding assays. We have exploited the unique properties of sdAb and used them as biological recognition elements in immuno-based sensors that can detect BoNT B.

Isolation and characterization of a thermally stable recombinant anti-caffeine heavy-chain antibody fragment.

PubMed

Ladenson, Ruth C; Crimmins, Dan L; Landt, Yvonne; Ladenson, Jack H

2006-07-01

We have isolated and characterized a caffeine-specific, heavy-chain-only antibody fragment (V(HH)) from llama that is capable of being utilized to analyze caffeine in hot and cold beverages. Camelid species (llama and camel) were selected for immunization because of their potential to make heat-stable, heavy-chain-only antibodies. Llamas and camels were immunized with caffeine covalently linked to keyhole limpet hemocyanin, and recombinant antibody techniques were used to create phage displayed libraries of variable region fragments of the heavy-chain antibodies. Caffeine-specific V(HH) fragments were selected by their ability to bind to caffeine/bovine serum albumin (BSA) and confirmed by a positive reaction in a caffeine enzyme-linked immunosorbent assay (caffeine ELISA). One of these V(HH) fragments (VSA2) was expressed as a soluble protein and shown to recover its reactivity after exposure to temperatures up to 90 degrees C. In addition, VSA2 was able to bind caffeine at 70 degrees C. A competition caffeine ELISA was developed for the measurement of caffeine in beverages, and concentrations of caffeine obtained for coffee, Coca-Cola Classic, and Diet Coke agreed well with high performance liquid chromatography (HPLC) determination and literature values. VSA2 showed minimal cross reactivity with structurally related methylxanthines.

Ovarian function in South American camelids (alpacas, llamas, vicunas, guanacos).

PubMed

Vaughan, Jane

2011-04-01

Ultrasound technology and hormone assays have provided a better understanding of folliculogenesis and ovulation in South American camelids in the last two decades. Females exhibit waves of ovarian follicular growth and are induced ovulators and therefore do not exhibit oestrous cycles in the manner of spontaneously ovulating species such as sheep and cattle. There is much variation in inter-wave interval among camelid species (alpaca/llama 10-22 days, vicuna 4-11 days), within species and within individual animals as the range of each phase of follicular growth is wide. Ovulation occurs 24-30h after mating and luteolysis occurs approximately 10 days later if conception fails to occur. Copyright © 2010 Elsevier B.V. All rights reserved.

Evaluation of intra-abdominal vasectomy in llamas and alpacas.

PubMed

Bravo, P W; Sumar, J

1991-11-01

A method of intra-abdominal vasectomy (deferentectomy), using a laparoscopic technique in llamas and alpacas, was evaluated. Food was withheld from 14 animals for 24 hours before sedation and laparoscopy. The ductus deferens was located close to the bladder, and a 2- to 3-cm section of ductus deferens was resected with the aid of a pair of hook scissors. The procedure was completed in 5 to 8 minutes, and animals did not have any adverse effects. Forty-five days after vasectomy, animals were used for detecting sexually receptive females or inducing ovulation during reproductive physiologic studies. The procedure did not impair libido and had no effect on male sexual behavior for many years. The technique is simpler, safer, and faster than the traditional external approach.

Collection and analysis of peritoneal fluid from healthy llamas and alpacas.

PubMed

Cebra, Christopher K; Tornquist, Susan J; Reed, Shannon K

2008-05-01

To describe a technique for abdominocentesis in camelids and report peritoneal fluid biochemical and cytologic findings from healthy llamas and alpacas. Prospective study. Animals-17 adult llamas and 5 adult alpacas. Right paracostal abdominocentesis was performed. Peritoneal fluid was collected by gravity flow into tubes containing potassium-EDTA for cell count and cytologic evaluation and lithium heparin for biochemical analysis. Blood samples were collected via jugular venipuncture into heparinized tubes at the same time. Cytologic components were quantified. Fluid pH and concentrations of total carbon dioxide, sodium, potassium, chloride, lactate, and glucose were compared between peritoneal fluid and venous blood. All but 3 camelids had peritoneal fluid cell counts of < 3,000 nucleated cells/microL, with < 2,000 neutrophils/microL and < 1,040 large mononuclear cells/microL. All but 1 had peritoneal fluid protein concentrations of > or = 2.5 g/dL. Peritoneal fluid of camelids generally contained slightly less glucose, lactate, and sodium and roughly equal concentrations of potassium and chloride as venous blood. Peritoneal fluid was collected safely from healthy camelids. Compared with blood, peritoneal fluid usually had a low cell count and protein concentration, but some individuals had higher values. Electrolyte concentrations resembled those found in blood. High cell counts and protein concentrations found in peritoneal fluid of some healthy camelids may overlap with values found in diseased camelids, complicating interpretation of peritoneal fluid values.

Oleander intoxication in New World camelids: 12 cases (1995-2006).

PubMed

Kozikowski, Tania A; Magdesian, K Gary; Puschner, Birgit

2009-08-01

To characterize the clinical and clinicopathologic effects and evaluate outcome associated with oleander toxicosis in New World camelids. Retrospective case series. 11 llamas and 1 alpaca. Medical records from a veterinary medical teaching hospital from January 1, 1995, to December 31, 2006, were reviewed. Records of all New World camelids that had detectable amounts of oleandrin in samples of serum, urine, or gastrointestinal fluid were included in the study. Descriptive statistics were used to evaluate the history, physical examination findings, clinicopathologic data, and outcome of affected camelids. 11 llamas and 1 alpaca met the inclusion criteria of the study. Either oleander plants were present where the camelids resided (n = 7) or oleander plant material was identified in the hay fed to the camelids (5). One llama was dead on arrival at the hospital, and another was euthanized upon admission because of financial concerns. Of the 10 treated camelids, 9 had evidence of acute renal failure, 7 had gastrointestinal signs, and 4 had cardiac dysrhythmias on initial evaluation. The overall mortality rate was 25%, but the mortality rate for the 10 camelids that were medically treated was 10%. In New World camelids, oleander intoxication was associated with a triad of clinical effects (ie, renal, gastrointestinal, and cardiovascular dysfunction). Oleander intoxication often represented a herd problem but carried a fair to good prognosis if treated promptly. Oleander toxicosis should be considered a differential diagnosis in sick camelids.

Reproductive physiology of the male camelid.

PubMed

Bravo, P W; Johnson, L W

1994-07-01

The physiology of reproduction with emphasis on endocrinology of llamas and alpacas is addressed. Information regarding male anatomy, puberty, testicular function, semen description, and sexual behavior is also included.

Efficient heterologous expression and secretion in Aspergillus oryzae of a llama variable heavy-chain antibody fragment V(HH) against EGFR.

PubMed

Okazaki, Fumiyoshi; Aoki, Jun-ichi; Tabuchi, Soichiro; Tanaka, Tsutomu; Ogino, Chiaki; Kondo, Akihiko

2012-10-01

We have constructed a filamentous fungus Aspergillus oryzae that secretes a llama variable heavy-chain antibody fragment (V(HH)) that binds specifically to epidermal growth factor receptor (EGFR) in a culture medium. A major improvement in yield was achieved by fusing the V(HH) with a Taka-amylase A signal sequence (sTAA) and a segment of 28 amino acids from the N-terminal region of Rhizopus oryzae lipase (N28). The yields of secreted, immunologically active anti-EGFR V(HH) reached 73.8 mg/1 in a Sakaguchi flask. The V(HH) fragments were released from the sTAA or N28 proteins by an indigenous A. oryzae protease during cultivation. The purified recombinant V(HH) fragment was specifically recognized and could bind to the EGFR with a high affinity.

Llama-derived single domain antibodies specific for Abrus agglutinin.

PubMed

Goldman, Ellen R; Anderson, George P; Zabetakis, Dan; Walper, Scott; Liu, Jinny L; Bernstein, Rachael; Calm, Alena; Carney, James P; O'Brien, Thomas W; Walker, Jennifer L; Garber, Eric A E

2011-11-01

Llama derived single domain antibodies (sdAb), the recombinantly expressed variable heavy domains from the unique heavy-chain only antibodies of camelids, were isolated from a library derived from llamas immunized with a commercial abrin toxoid preparation. Abrin is a potent toxin similar to ricin in structure, sequence and mechanism of action. The selected sdAb were evaluated for their ability to bind to commercial abrin as well as abrax (a recombinant abrin A-chain), purified abrin fractions, Abrus agglutinin (a protein related to abrin but with lower toxicity), ricin, and unrelated proteins. Isolated sdAb were also evaluated for their ability to refold after heat denaturation and ability to be used in sandwich assays as both capture and reporter elements. The best binders were specific for the Abrus agglutinin, showing minimal binding to purified abrin fractions or unrelated proteins. These binders had sub nM affinities and regained most of their secondary structure after heating to 95 °C. They functioned well in sandwich assays. Through gel analysis and the behavior of anti-abrin monoclonal antibodies, we determined that the commercial toxoid preparation used for the original immunizations contained a high percentage of Abrus agglutinin, explaining the selection of Abrus agglutinin binders. Used in conjunction with anti-abrin monoclonal and polyclonal antibodies, these reagents can fill a role to discriminate between the highly toxic abrin and the related, but much less toxic, Abrus agglutinin and distinguish between different crude preparations.

Llama-Derived Single Domain Antibodies Specific for Abrus Agglutinin

PubMed Central

Goldman, Ellen R.; Anderson, George P.; Zabetakis, Dan; Walper, Scott; Liu, Jinny L.; Bernstein, Rachael; Calm, Alena; Carney, James P.; O’Brien, Thomas W.; Walker, Jennifer L.; Garber, Eric A. E.

2011-01-01

Llama derived single domain antibodies (sdAb), the recombinantly expressed variable heavy domains from the unique heavy-chain only antibodies of camelids, were isolated from a library derived from llamas immunized with a commercial abrin toxoid preparation. Abrin is a potent toxin similar to ricin in structure, sequence and mechanism of action. The selected sdAb were evaluated for their ability to bind to commercial abrin as well as abrax (a recombinant abrin A-chain), purified abrin fractions, Abrus agglutinin (a protein related to abrin but with lower toxicity), ricin, and unrelated proteins. Isolated sdAb were also evaluated for their ability to refold after heat denaturation and ability to be used in sandwich assays as both capture and reporter elements. The best binders were specific for the Abrus agglutinin, showing minimal binding to purified abrin fractions or unrelated proteins. These binders had sub nM affinities and regained most of their secondary structure after heating to 95 °C. They functioned well in sandwich assays. Through gel analysis and the behavior of anti-abrin monoclonal antibodies, we determined that the commercial toxoid preparation used for the original immunizations contained a high percentage of Abrus agglutinin, explaining the selection of Abrus agglutinin binders. Used in conjunction with anti-abrin monoclonal and polyclonal antibodies, these reagents can fill a role to discriminate between the highly toxic abrin and the related, but much less toxic, Abrus agglutinin and distinguish between different crude preparations. PMID:22174977

Mating induces production of MMP2 in the llama oviduct: Analysis of MMP2 effect on semen.

PubMed

Zampini, Renato; Sari, Luciana M; Argañaraz, Martin E; Fumuso, Fernanda G; Barraza, Daniela E; Carretero, María I; Apichela, Silvana A

2018-05-01

Ovulation of South American Camelids is induced by mating. After copulation, sperm are stored into the oviduct to be released near ovulation time. To study whether copulation induces matrix metalloproteinase-2 (MMP2) secretion in the oviduct, the occurrence of MMP2 in oviductal tissue and oviductal fluid (OF) from 24 h post-mated was compared with non-mated llama females. There was an incremental increase of MMP2 in the oviductal epithelial cells, and MMP2 activity in OF after copulation. Additionally, MMP2 activator (MMP14), inducer (EMMPRIN) and inhibitor (TIMP2) were present in the oviductal epithelial cells of both non-mated and post-mated females. A post-mating segment-specific regulation occurred because relative abundance of TIMP2 mRNA was greater in the utero tubal-junction which was accompanied with a reduced amount of MMP14 in the ampulla in comparison with the non-mated females. To examine the effect of MMP2 on semen liquefaction and sperm physiology, the effects of addition of recombinant human MMP2 was evaluated. The MMP2 had no effect on semen thread formation and seminal plasma protein profile. Sperm viability and plasma membrane function were not influenced by the enzyme treatment either. In summary, in llamas the oviductal microenvironment changes in response to stimuli induced by copulation, increasing the production and secretion of MMP2. Copyright © 2018 Elsevier B.V. All rights reserved.

16 CFR 300.18 - Use of name of specialty fiber.

Code of Federal Regulations, 2013 CFR

2013-01-01

... designation permitted under this rule: 55% Alpaca—45% Camel Hair 50% Recycled Camel Hair—50% Wool 60% Recycled Alpaca—40% Rayon 35% Recycled Llama—35% Recycled Vicuna—30% Cotton 60% Cotton—40% Recycled Llama. (b...

16 CFR 300.18 - Use of name of specialty fiber.

Code of Federal Regulations, 2014 CFR

2014-01-01

... designation permitted under this rule: 55% Alpaca—45% Camel Hair 50% Recycled Camel Hair—50% Wool 60% Recycled Alpaca—40% Rayon 35% Recycled Llama—35% Recycled Vicuna—30% Cotton 60% Cotton—40% Recycled Llama. (b...

16 CFR 300.18 - Use of name of specialty fiber.

Code of Federal Regulations, 2010 CFR

2010-01-01

... designation permitted under this rule: 55% Alpaca—45% Camel Hair 50% Recycled Camel Hair—50% Wool 60% Recycled Alpaca—40% Rayon 35% Recycled Llama—35% Recycled Vicuna—30% Cotton 60% Cotton—40% Recycled Llama. (b...

16 CFR 300.18 - Use of name of specialty fiber.

Code of Federal Regulations, 2012 CFR

2012-01-01

... designation permitted under this rule: 55% Alpaca—45% Camel Hair 50% Recycled Camel Hair—50% Wool 60% Recycled Alpaca—40% Rayon 35% Recycled Llama—35% Recycled Vicuna—30% Cotton 60% Cotton—40% Recycled Llama. (b...

[South American camelids in Switzerland. II. Reference values for blood parameters].

PubMed

Hengrave Burri, I; Tschudi, P; Martig, J; Liesegang, A; Meylan, M

2005-08-01

In order to establish reference values for blood parameters of South American camelids in Switzerland, 273 blood samples were collected from 141 llamas and 132 alpacas. These animals were classified in three categories (young animals < six months, adult females and males). Forty-one parameters were measured (red blood cell count, white blood cell count, electrolytes, metabolites and enzymes). Significant differences between llamas and alpacas were evident for 26 parameters. This study also showed that differences between young animals, females and males must be taken into consideration. A comparison of blood values with the results of fecal analysis for parasite eggs showed that an infestation with Dicrocoelium dendriticum was associated with elevated activity of two liver enzymes (GLDH and gamma-GT) in the serum. In contrast, no differences were found in the results of blood analyses between animals shedding eggs of gastrointestinal strongyles or not.

Characterization of hypertriglyceridemia and response to treatment with insulin in llamas and alpacas: 31 cases (1995-2005).

PubMed

Waitt, Laura H; Cebra, Christopher K

2008-05-01

To evaluate camelids with hypertriglyceridemia with regard to signalment, clinical features of disease, and response to treatment with insulin. Retrospective case series. 23 alpacas and 8 llamas with hypertriglyceridemia. For analysis of medical record data, 20 hypertriglyceridemic camelids with multiple recorded measurements of serum or plasma triglycerides concentration were classified as follows: those with an initial triglycerides concentration > 60 to > or = 500 mg/dL that were or were not treated with insulin (HT-I and HT-N camelids, respectively) and those with an initial triglycerides concentration > 500 mg/dL that were treated with insulin (lipemic [LIP-I] camelids). Only 1 recorded triglycerides concentration was available for an additional 11 hypertriglyceridemic camelids; data from those records were included in the characterization of signalment and clinical features of disease. Compared with the general population of hospitalized camelids, hypertriglyceridemic camelids did not differ significantly with respect to age or sex. Of 22 female camelids, only 7 were lactating or pregnant. Serum or plasma triglycerides concentrations in HT-N and HT-I camelids did not differ significantly at admission, but triglycerides concentrations in HT-I camelids decreased significantly after insulin treatment. Posttreatment triglycerides concentrations in HT-I camelids were significantly lower than those in HT-N camelids. During the period of hospitalization, triglycerides concentrations in HT-N camelids increased, whereas those in LIP-I camelids decreased significantly. Results indicated that hypertriglyceridemia affects llamas and alpacas of all ages and both sexes. Insulin treatment may reduce serum or plasma triglycerides concentrations in camelids with hypertriglyceridemia.

The postpartum llama: fertility after parturition.

PubMed

Bravo, P W; Fowler, M E; Lasley, B L

1994-12-01

Fertility was evaluated at various times during the postpartum period in the llama. Fifty-six parous female llamas chosen at random were bred at 10, 20, and 30 days postpartum with six intact males. Half of the females copulated only once and the other half twice within an interval of 24 h. Ovarian activity was monitored by ultrasonography and analysis of urinary estrone sulfate and pregnanediol glucuronide (PdG). At the time of copulation, all females had developed ovulatory-size follicles. Ovulation was confirmed by the presence of a CL at 8 days after breeding and PdG concentrations > 1 ng/mg creatinine (Cr). Conception was defined as PdG concentrations > 1 ng/mg Cr at 15 days after breeding and the presence of a CL. Pregnancy was defined as the presence of a CL, PdG concentrations > 1 ng/mg Cr and an embryonic vesicle 20 days postbreeding. There were no significant differences in the proportion of females ovulating after breeding at different times postpartum; however, conception and pregnancy were significantly greater in females bred at Day 20 or 30. Ovarian follicle size was significantly larger at 30 days (9.1 mm) than at 10 and 20 days (7.9 and 8.8 mm, respectively) of breeding, with no difference in concentrations of estrone sulfate. The proportion of females conceiving as a result of breeding at 10 days postpartum (6 out of 10) was significantly less than for females bred at 20 (13 out of 15) or 30 days (16 out of 18).(ABSTRACT TRUNCATED AT 250 WORDS)

Sarcocystis masoni, n. sp. (Apicomplexa: Sarcocystidae), and redescription of Sarcocystis aucheniae from llama (Lama glama), guanaco (Lama guanicoe) and alpaca (Vicugna pacos).

PubMed

Moré, Gastón; Regensburger, Cristian; Gos, M Laura; Pardini, Lais; Verma, Shiv K; Ctibor, Juliana; Serrano-Martínez, Marcos Enrique; Dubey, Jitender P; Venturini, M Cecilia

2016-04-01

There is considerable confusion concerning the species of Sarcocystis in South American camelids (SAC). Several species names have been used; however, proper descriptions are lacking. In the present paper, we redescribe the macroscopic sarcocyst forming Sarcocystis aucheniae and describe and propose a new name, Sarcocystis masoni for the microscopic sarcocyst forming species. Muscles samples were obtained from llamas (Lama glama) and guanacos (Lama guanicoe) from Argentina and from alpacas (Vicugna pacos) and llamas from Peru. Individual sarcocysts were processed by optical and electron microscopy, and molecular studies. Microscopic sarcocysts of S. masoni were up to 800 µm long and 35-95 µm wide, the sarcocyst wall was 2·5-3·5 µm thick, and had conical to cylindrical villar protrusions (vp) with several microtubules. Each vp had 11 or more rows of knob-like projections. Seven 18S rRNA gene sequences obtained from sarcocysts revealed 95-96% identity with other Sarcocystis spp. sequences reported in the GenBank. Sarcocysts of S. aucheniae were macroscopic, up to 1·2 cm long and surrounded by a dense and laminar 50 µm thick secondary cyst wall. The sarcocyst wall was up to 10 µm thick, and had branched vp, appearing like cauliflower. Comparison of the 11 sequences obtained from individual macroscopic cysts evidenced a 98-99% of sequence homology with other S. aucheniae sequences. In conclusion, 2 morphologically and molecularly different Sarcocystis species, S. masoni (microscopic cysts) and S. aucheniae (macroscopic cysts), were identified affecting different SAC from Argentina and Peru.

Bioavailability and pharmacokinetics of oral meloxicam in llamas.

PubMed

Kreuder, Amanda J; Coetzee, Johann F; Wulf, Larry W; Schleining, Jennifer A; KuKanich, Butch; Layman, Lori L; Plummer, Paul J

2012-06-21

South American camelids in the United States have rapidly developed into an important agricultural industry in need of veterinary services. Pain management is challenging in camelids because there are no drugs currently approved by the U.S. Food and Drug Administration for use in these species. Dosage regimens used for many therapeutic drugs have been extrapolated from other ruminants; however, the pharmacokinetics, in camelids, may differ from those of other species. Studies investigating the pharmacokinetics of cyclooxygenase-2 (COX-2) selective non-steroidal anti-inflammatory drugs in camelids are deficient in the published literature. Six adult llamas (121- 168 kg) were administered either a 1 mg/kg dose of oral or a 0.5 mg/kg dose of IV meloxicam in a randomized cross-over design with an 11 day washout period between treatments. Plasma samples collected up to 96 hours post-administration were analyzed by high pressure liquid chromatography and mass spectrometry detection (HPLC-MS) followed by non-compartmental pharmacokinetic analysis. A mean peak plasma concentration (CMAX) of 1.314 μg/mL (Range: 0.826 - 1.776 μg/mL) was recorded at 21.4 hours (Range: 12.0 - 24.0 hours) with a half-life (T ½ λz) of 22.7 hours (Range: 18.0 - 30.8 hours) after oral meloxicam administration. In comparison, a half-life (T ½ λz) of 17.4 hours (Range: 16.2 - 20.7 hours) was demonstrated with IV meloxicam administration. The oral bioavailability (F) of meloxicam (dose normalized) was 76% (Range: 48 - 92%). No adverse effects associated with either treatment modality were observed in the llamas. The mean bioavailability (F) of oral meloxicam was 76% indicating a high degree of gastrointestinal absorption. Plasma meloxicam concentrations >0.2 μg/mL were maintained for up to 72 h after oral administration; >0.2 μg/mL is considered to be the concentration of meloxicam required for analgesic effects in other species such as the horse. These data suggest that a single dosage of oral meloxicam at 1 mg/kg could potentially maintain therapeutic concentrations in plasma for up to 3 days in adult llamas.

Adaptive functional specialisation of architectural design and fibre type characteristics in agonist shoulder flexor muscles of the llama, Lama glama

PubMed Central

Graziotti, Guillermo H; Chamizo, Verónica E; Ríos, Clara; Acevedo, Luz M; Rodríguez-Menéndez, J M; Victorica, C; Rivero, José-Luis L

2012-01-01

Like other camelids, llamas (Lama glama) have the natural ability to pace (moving ipsilateral limbs in near synchronicity). But unlike the Old World camelids (bactrian and dromedary camels), they are well adapted for pacing at slower or moderate speeds in high-altitude habitats, having been described as good climbers and used as pack animals for centuries. In order to gain insight into skeletal muscle design and to ascertain its relationship with the llama’s characteristic locomotor behaviour, this study examined the correspondence between architecture and fibre types in two agonist muscles involved in shoulder flexion (M. teres major – TM and M. deltoideus, pars scapularis – DS and pars acromialis – DA). Architectural properties were found to be correlated with fibre-type characteristics both in DS (long fibres, low pinnation angle, fast-glycolytic fibre phenotype with abundant IIB fibres, small fibre size, reduced number of capillaries per fibre and low oxidative capacity) and in DA (short fibres, high pinnation angle, slow-oxidative fibre phenotype with numerous type I fibres, very sparse IIB fibres, and larger fibre size, abundant capillaries and high oxidative capacity). This correlation suggests a clear division of labour within the M. deltoideus of the llama, DS being involved in rapid flexion of the shoulder joint during the swing phase of the gait, and DA in joint stabilisation during the stance phase. However, the architectural design of the TM muscle (longer fibres and lower fibre pinnation angle) was not strictly matched with its fibre-type characteristics (very similar to those of the postural DA muscle). This unusual design suggests a dual function of the TM muscle both in active flexion of the shoulder and in passive support of the limb during the stance phase, pulling the forelimb to the trunk. This functional specialisation seems to be well suited to a quadruped species that needs to increase ipsilateral stability of the limb during the support phase of the pacing gait. Compared with other species, llama skeletal muscles are well suited for greater force generation combined with higher fatigue resistance during exercise. These characteristics are interpreted as being of high adaptive value, given the llama’s habitat and its use as a pack animal. PMID:22625659

25 CFR 161.1 - What definitions do I need to know?

Code of Federal Regulations, 2014 CFR

2014-04-01

... following: (1) One adult sheep or goat is equivalent to one-fifth (0.20) of an AU; (2) One adult horse, mule..., cattle, sheep, goats, mules, burros, donkeys, and llamas. Management Unit is a subdivision of a...

25 CFR 161.1 - What definitions do I need to know?

Code of Federal Regulations, 2012 CFR

2012-04-01

... following: (1) One adult sheep or goat is equivalent to one-fifth (0.20) of an AU; (2) One adult horse, mule..., cattle, sheep, goats, mules, burros, donkeys, and llamas. Management Unit is a subdivision of a...

25 CFR 161.1 - What definitions do I need to know?

Code of Federal Regulations, 2011 CFR

2011-04-01

... following: (1) One adult sheep or goat is equivalent to one-fifth (0.20) of an AU; (2) One adult horse, mule..., cattle, sheep, goats, mules, burros, donkeys, and llamas. Management Unit is a subdivision of a...

Clinical pathology of llamas and alpacas.

PubMed

Tornquist, Susan J

2009-07-01

Clinical laboratory data including hematology, hemostasis, biochemical, and cytologic findings contribute to diagnosis and monitoring of numerous conditions in camelids. Establishment of reference intervals and descriptions of normal components of fluids in these species have improved our ability to interpret test results.

The fine structure of the stomach mucosa of the Llama (Llama guanacoe). II. The fundic region of the hind stomach.

PubMed

Luciano, L; Reale, E; von Engelhardt, W

1980-01-01

The epithelium of the fundic region mucosa of the hind stomach in the Llama guanacoe has been studied using morphological and histochemical methods. Morphology suggests that solute and water absorption may occur in the epithelium of the surface and of the foveolae, although this absorption can not be estimated because of the extensive secretion of the gastric glands. The same cells of the surface and foveolar epithelium show numerous secretory granules. The glands reveal neck cells, chief cells, a large number of oxyntic cells, four types of endocrine cells (A-like, ECL, D and EC), brush cells and wandering cells. PAS and Alcian blue reactions for light microscopy suggest a secretion of neutral and acidic mucosubstances in the surface and foveolar epithelium, of neutral mucosubstances only in the neck cells. Periodic acid-thiocarbohydrazide silver proteinate (PA-TCH-SP) reaction for electron microscopy confirms the presence of neutral mucosubstances within the secretory granules of the surface, foveolar and neck epithelial cells. In all these cells, the reaction product is also evident within sacculi and vesicles of the maturing surface of the Golgi apparatus. A positive PA-TCH-SP reaction also occurs on the membrane (and not on the contents) of the Golgi apparatus (maturing surface) and of the secretory granules of the chief cells as well as on the membrane of the Golgi apparatus and of apical vesicles and tubules of the oxyntic cells. In addition, silver granules slightly enhance the electron desity of the contents of the secretory granules in the endocrine cells. Morphological and histochemical findings are discussed and compared with results described by others for monogastric mammals.

Molecular Characterization of the Llamas (Lama glama) Casein Cluster Genes Transcripts (CSN1S1, CSN2, CSN1S2, CSN3) and Regulatory Regions

PubMed Central

Pauciullo, Alfredo; Erhardt, Georg

2015-01-01

In the present paper, we report for the first time the characterization of llama (Lama glama) caseins at transcriptomic and genetic level. A total of 288 casein clones transcripts were analysed from two lactating llamas. The most represented mRNA populations were those correctly assembled (85.07%) and they encoded for mature proteins of 215, 217, 187 and 162 amino acids respectively for the CSN1S1, CSN2, CSN1S2 and CSN3 genes. The exonic subdivision evidenced a structure made of 21, 9, 17 and 6 exons for the αs1-, β-, αs2- and κ-casein genes respectively. Exon skipping and duplication events were evidenced. Two variants A and B were identified in the αs1-casein gene as result of the alternative out-splicing of the exon 18. An additional exon coding for a novel esapeptide was found to be cryptic in the κ-casein gene, whereas one extra exon was found in the αs2-casein gene by the comparison with the Camelus dromedaries sequence. A total of 28 putative phosphorylated motifs highlighted a complex heterogeneity and a potential variable degree of post-translational modifications. Ninety-six polymorphic sites were found through the comparison of the lama casein cDNAs with the homologous camel sequences, whereas the first description and characterization of the 5’- and 3’-regulatory regions allowed to identify the main putative consensus sequences involved in the casein genes expression, thus opening the way to new investigations -so far- never achieved in this species. PMID:25923814

Molecular Characterization of the Llamas (Lama glama) Casein Cluster Genes Transcripts (CSN1S1, CSN2, CSN1S2, CSN3) and Regulatory Regions.

PubMed

Pauciullo, Alfredo; Erhardt, Georg

2015-01-01

In the present paper, we report for the first time the characterization of llama (Lama glama) caseins at transcriptomic and genetic level. A total of 288 casein clones transcripts were analysed from two lactating llamas. The most represented mRNA populations were those correctly assembled (85.07%) and they encoded for mature proteins of 215, 217, 187 and 162 amino acids respectively for the CSN1S1, CSN2, CSN1S2 and CSN3 genes. The exonic subdivision evidenced a structure made of 21, 9, 17 and 6 exons for the αs1-, β-, αs2- and κ-casein genes respectively. Exon skipping and duplication events were evidenced. Two variants A and B were identified in the αs1-casein gene as result of the alternative out-splicing of the exon 18. An additional exon coding for a novel esapeptide was found to be cryptic in the κ-casein gene, whereas one extra exon was found in the αs2-casein gene by the comparison with the Camelus dromedaries sequence. A total of 28 putative phosphorylated motifs highlighted a complex heterogeneity and a potential variable degree of post-translational modifications. Ninety-six polymorphic sites were found through the comparison of the lama casein cDNAs with the homologous camel sequences, whereas the first description and characterization of the 5'- and 3'-regulatory regions allowed to identify the main putative consensus sequences involved in the casein genes expression, thus opening the way to new investigations -so far- never achieved in this species.

A review of coccidiosis in South American camelids

USDA-ARS?s Scientific Manuscript database

Camelids (llama, alpaca, vicuñas, guanacos) are important for the economy of South America and Eimeria infections are important as cause of mortality in camelids. Of the five valid species of Eimeria in South American camelids, Eimeria macusaniensis, Eimeria lamae, Eimeria alpacae, Eimeria punoensis...

7 CFR 760.304 - Covered livestock.

Code of Federal Regulations, 2010 CFR

2010-01-01

..., adult or non-adult dairy cattle, alpacas, deer, elk, emus, equine, goats, llamas, poultry, reindeer... current production year; (4) Been maintained for commercial use as part of the producer's farming... for reasons other than commercial use as part of a farming operation. Such excluded uses include, but...

High production of llama variable heavy-chain antibody fragment (VHH) fused to various reader proteins by Aspergillus oryzae.

PubMed

Hisada, Hiromoto; Tsutsumi, Hiroko; Ishida, Hiroki; Hata, Yoji

2013-01-01

Llama variable heavy-chain antibody fragment (VHH) fused to four different reader proteins was produced and secreted in culture medium by Aspergillus oryzae. These fusion proteins consisted of N-terminal reader proteins, VHH, and a C-terminal his-tag sequence which facilitated purification using one-step his-tag affinity chromatography. SDS-PAGE analysis of the deglycosylated purified fusion proteins confirmed that the molecular weight of each corresponded to the expected sum of VHH and the respective reader proteins. The apparent high molecular weight reader protein glucoamylase (GlaB) was found to be suitable for efficient VHH production. The GlaB-VHH-His protein bound its antigen, human chorionic gonadotropin, and was detectable by a new ELISA-based method using a coupled assay with glucoamylase, glucose oxidase, peroxidase, maltose, and 3,3',5,5'-tetramethylbenzidine as substrates. Addition of potassium phosphate to the culture medium induced secretion of 0.61 mg GlaB-VHH-His protein/ml culture medium in 5 days.

Clinical, ultrasonographic, and laboratory findings in 12 llamas and 12 alpacas with malignant round cell tumors

PubMed Central

Martin, Jeanne M.; Valentine, Beth A.; Cebra, Christopher K.

2010-01-01

Clinical signs, duration of illness, clinicopathologic findings, and ultrasonographic findings were evaluated in 12 llamas and 12 alpacas with malignant round cell tumors (MRCT). All but 1 animal died or was euthanized. Common clinical findings were anorexia, recumbency or weakness, and weight loss or poor growth. Peripheral lymphadenomegaly occurred in only 7 animals and was detected more often at necropsy than during physical examination. Common clinicopathologic abnormalities were hypoalbuminemia, acidosis, azotemia, anemia, hyperglycemia, and neutrophilia. Ultrasonography detected tumors in 4/6 animals. Cytologic evaluation of fluid or tissue aspirates or histopathology of biopsy tissue was diagnostic in 5/6 cases. A clinical course of 2 wk or less prior to death or euthanasia was more common in animals ≤ 2 y of age (9/11) than in older animals (6/13). Regular examination of camelids to include clinical pathology and evaluation of peripheral lymph nodes may result in early detection of MCRT. PMID:21358931

Zygomycetes From Herbivore Dung in the Ecological Reserve of Dois IrmÃOs, Northeast Brazil

PubMed Central

de Azevedo Santiago, André Luiz Cabral Monteiro; Botelho Trufem, Sandra Farto; Malosso, Elaine; dos Santos, Paulo Jorge Parreira; de Queiroz Cavalcanti, Maria Auxiliadora

2011-01-01

Thirty-eight taxa of Zygomycetes distributed in 15 genera were recorded from tapir (Tapirus terrestris), camel (Camelus bactrianus), horse (Equus caballus), deer (Cervus elaphus), agouti (Dasyprocta aguti), donkey (Equus asinus), llama (Llama glama) and waterbuck (Kobus ellipsiprymnus) dung collected at the Reserva Ecológica de Dois Irmãos located in Recife, State of Pernambuco, Northeast Brazil. The samples were collected on a monthly basis from June 2005 to May 2006, taken to the laboratory and incubated in moist chambers. Higher number of taxa was observed in the excrements of tapir, followed by deer and donkey. The highest number of species was detected for Mucor, followed by Pilobolus. Statistical analyses showed significant differences in richness of Zygomycetes taxa between the herbivore dung types. Differences of species composition, however, were weak. Seasonality influenced the Zygomycetes species composition but not its richness. Variations in taxa composition between ruminants and non-ruminants dung were non significant. PMID:24031609

Zygomycetes From Herbivore Dung in the Ecological Reserve of Dois IrmÃOs, Northeast Brazil.

PubMed

de Azevedo Santiago, André Luiz Cabral Monteiro; Botelho Trufem, Sandra Farto; Malosso, Elaine; Dos Santos, Paulo Jorge Parreira; de Queiroz Cavalcanti, Maria Auxiliadora

2011-01-01

Thirty-eight taxa of Zygomycetes distributed in 15 genera were recorded from tapir (Tapirus terrestris), camel (Camelus bactrianus), horse (Equus caballus), deer (Cervus elaphus), agouti (Dasyprocta aguti), donkey (Equus asinus), llama (Llama glama) and waterbuck (Kobus ellipsiprymnus) dung collected at the Reserva Ecológica de Dois Irmãos located in Recife, State of Pernambuco, Northeast Brazil. The samples were collected on a monthly basis from June 2005 to May 2006, taken to the laboratory and incubated in moist chambers. Higher number of taxa was observed in the excrements of tapir, followed by deer and donkey. The highest number of species was detected for Mucor, followed by Pilobolus. Statistical analyses showed significant differences in richness of Zygomycetes taxa between the herbivore dung types. Differences of species composition, however, were weak. Seasonality influenced the Zygomycetes species composition but not its richness. Variations in taxa composition between ruminants and non-ruminants dung were non significant.

No Rhyme or Reason.

ERIC Educational Resources Information Center

Flannery, Maura C.

1995-01-01

Contains a summary of the latest research findings related to the topics of eye development, mimicry, evolution of llamas, hybridization between Arctic wolves and huskies, ultraviolet light and algae, acid rock, orchids and funerals, enzymes that can withstand heat, fungus-killing compounds, chaos theory and the treatment of epileptics, sleep and…

19 CFR 10.244 - Certificate of Origin.

Code of Federal Regulations, 2011 CFR

2011-04-01

... assembled from U.S. formed, dyed, printed and finished fabrics or fabric components, or U.S. formed knit-to... llama, alpaca or vicuña fabrics, fabric components, or knit-to-shape components, from Andean yarns 10... more yarns, fabrics, fabric components, or knit-to-shape components described in preference groups A...

19 CFR 10.244 - Certificate of Origin.

Code of Federal Regulations, 2010 CFR

2010-04-01

... assembled from U.S. formed, dyed, printed and finished fabrics or fabric components, or U.S. formed knit-to... llama, alpaca or vicuña fabrics, fabric components, or knit-to-shape components, from Andean yarns 10... more yarns, fabrics, fabric components, or knit-to-shape components described in preference groups A...

Single-Domain Antibodies: Rugged Recognition Element

DTIC Science & Technology

2007-01-01

and spiny dogfish shark (60 million representatives), which are displayed on filamentous phage M13 . We selected the llama- derived library for binding...subject to a penalty for failing to comply with a collection of information if it does not display a currently valid OMB control number. 1. REPORT DATE

Imaging diagnosis--pulmonary metastases in New World camelids.

PubMed

Gall, David A; Zekas, Lisa J; Van Metre, David; Holt, Timothy

2006-01-01

The radiographic appearance of pulmonary metastatic disease from carcinoma is described in a llama and an alpaca. In one, a diffuse miliary pattern was seen. In the other, a more atypical unstructured interstitial pattern was recognized. Metastatic pulmonary neoplasia in camelids may assume a generalized miliary or unstructured pattern.

Llama Herders and Urban Elites: Interdisciplinary Readings of Early Colonial Narratives in the Americas

ERIC Educational Resources Information Center

Beaule, Christine D.; Quintana, Benito

2017-01-01

We argue for an interdisciplinary pedagogical approach that we call the Integration of Research and Education in the Classroom, which highlights and crosses disciplinary boundaries to challenge each field's assumptions, limitations, conceptual and interpretive purview. We use a set of examples that center on problematizing various aspects of the…

7 CFR 760.204 - Eligible livestock, honeybees, and farm-raised fish.

Code of Federal Regulations, 2010 CFR

2010-01-01

..., goats, llamas, poultry, reindeer, sheep, or swine; (2) Be livestock that would normally have been... livestock that has been maintained for commercial use as part of the producer's farming operation on the... been produced and maintained for reasons other than commercial use as part of a farming operation; and...

25 CFR 161.1 - What definitions do I need to know?

Code of Federal Regulations, 2010 CFR

2010-04-01

... following: (1) One adult sheep or goat is equivalent to one-fifth (0.20) of an AU; (2) One adult horse, mule..., cattle, sheep, goats, mules, burros, donkeys, and llamas. Management Unit is a subdivision of a... management plan is a statement of management objectives for grazing, farming, or other agriculture management...

Antibody engineering reveals the important role of J segments in the production efficiency of llama single-domain antibodies in Saccharomyces cerevisiae.

PubMed

Gorlani, A; Hulsik, D Lutje; Adams, H; Vriend, G; Hermans, P; Verrips, T

2012-01-01

Variable domains of llama heavy-chain antibodies (VHH) are becoming a potent tool for a wide range of biotechnological and medical applications. Because of structural features typical of their single-domain nature, they are relatively easy to produce in lower eukaryotes, but it is not uncommon that some molecules have poor secretion efficiency. We therefore set out to study the production yield of VHH. We computationally identified five key residues that are crucial for folding and secretion, and we validated their importance with systematic site-directed mutations. The observation that all key residues were localised in the V segment, in proximity of the J segment of VHH, led us to study the importance of J segment in secretion efficiency. Intriguingly, we found that the use of specific J segments in VHH could strongly influence the production yield. Sequence analysis and expression experiments strongly suggested that interactions with chaperones, especially with the J segment, are a crucial aspect of the production yield of VHH.

Oxygen transfer properties and dimensions of red blood cells in high-altitude camelids, dromedary camel and goat.

PubMed

Yamaguchi, K; Jürgens, K D; Bartels, H; Piiper, J

1987-01-01

To estimate the advantage of the small red blood cells (RBC) of high-altitude camelids for O2 transfer, the kinetics of O2 uptake into and release from the RBC obtained from llama, vicuña and alpaca were investigated at 37 degrees C with a stopped-flow technique. O2 transfer conductance of RBC (G) was estimated from the rate of O2 saturation change and the corresponding O2 pressure difference between medium and hemoglobin. For comparison, O2 kinetics for the RBC of a low-altitude camelid (dromedary camel) and the pygmy goat were determined and previously measured values for human RBC were used. O2 transfer of RBC was found to be strongly influenced by extracellular diffusion, except with O2 release into dithionite solutions of sufficiently high concentration (greater than 30 mM). The G values measured in these 'standard' conditions, Gst (in mmol X min-1 X Torr-1 X (ml RBC)-1) were: high-altitude camelids, 0.58 (averaged for llama, alpaca and vicuña since there were no significant interspecific differences); camel 0.42; goat, 0.42; man, 0.39. The differences can in part be attributed to expected effects of the size and shape of the RBC (volume, surface area, mean thickness), as well as to the intracellular O2 diffusivity which depends on the concentration of cellular hemoglobin. The high Gst of RBC of high-altitude camelids may be considered to enhance O2 transfer in lungs and tissues. But the O2 transfer conductance of blood, theta, equal to Gst multiplied by hematocrit (in mmol X min-1 X Torr-1 X (ml blood)-1), was only slightly higher as compared to other species: 0.20 (llama, alpaca, vicuña), 0.14 (camel), 0.18 (goat), 0.17 (man).

Llama Antibody Fragments Recognizing Various Epitopes of the CD4bs Neutralize a Broad Range of HIV-1 Subtypes A, B and C

PubMed Central

Aasa-Chapman, Marlèn; Gorlani, Andrea; Forsman Quigley, Anna; Hulsik, David Lutje; Chen, Lei; Weiss, Robin; de Haard, Hans; Verrips, Theo

2012-01-01

Many of the neutralising antibodies, isolated to date, display limited activities against the globally most prevalent HIV-1 subtypes A and C. Therefore, those subtypes are considered to be an important target for antibody-based therapy. Variable domains of llama heavy chain antibodies (VHH) have some superior properties compared with classical antibodies. Therefore we describe the application of trimeric forms of envelope proteins (Env), derived from HIV-1 of subtype A and B/C, for a prolonged immunization of two llamas. A panel of VHH, which interfere with CD4 binding to HIV-1 Env were selected with use of panning. The results of binding and competition assays to various Env, including a variant with a stabilized CD4-binding state (gp120Ds2), cross-competition experiments, maturation analysis and neutralisation assays, enabled us to classify the selected VHH into three groups. The VHH of group I were efficient mainly against viruses of subtype A, C and B′/C. The VHH of group II resemble the broadly neutralising antibody (bnmAb) b12, neutralizing mainly subtype B and C viruses, however some had a broader neutralisation profile. A representative of the third group, 2E7, had an even higher neutralization breadth, neutralizing 21 out of the 26 tested strains belonging to the A, A/G, B, B/C and C subtypes. To evaluate the contribution of certain amino acids to the potency of the VHH a small set of the mutants were constructed. Surprisingly this yielded one mutant with slightly improved neutralisation potency against 92UG37.A9 (subtype A) and 96ZM651.02 (subtype C). These findings and the well-known stability of VHH indicate the potential application of these VHH as anti-HIV-1 microbicides. PMID:22438910

A Single-Domain Llama Antibody Potently Inhibits the Enzymatic Activity of Botulinum Neurotoxin by Binding to the Non-Catalytic [alpha]-Exosite Binding Region

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dong, Jianbo; Thompson, Aaron A.; Fan, Yongfeng

2010-08-13

Ingestion or inhalation of botulinum neurotoxin (BoNT) results in botulism, a severe and frequently fatal disease. Current treatments rely on antitoxins, which, while effective, cannot reverse symptoms once BoNT has entered the neuron. For treatments that can reverse intoxication, interest has focused on developing inhibitors of the enzymatic BoNT light chain (BoNT Lc). Such inhibitors typically mimic substrate and bind in or around the substrate cleavage pocket. To explore the full range of binding sites for serotype A light chain (BoNT/A Lc) inhibitors, we created a library of non-immune llama single-domain VHH (camelid heavy-chain variable region derived from heavy-chain-only antibody)more » antibodies displayed on the surface of the yeast Saccharomyces cerevisiae. Library selection on BoNT/A Lc yielded 15 yeast-displayed VHH with equilibrium dissociation constants (K{sub d}) from 230 to 0.03 nM measured by flow cytometry. Eight of 15 VHH inhibited the cleavage of substrate SNAP25 (synaptosome-associated protein of 25,000 Da) by BoNT/A Lc. The most potent VHH (Aa1) had a solution K{sub d} for BoNT/A Lc of 1.47 x 10{sup -10} M and an IC{sub 50} (50% inhibitory concentration) of 4.7 x 10{sup -10} M and was resistant to heat denaturation and reducing conditions. To understand the mechanism by which Aa1 inhibited catalysis, we solved the X-ray crystal structure of the BoNT/A Lc-Aa1 VHH complex at 2.6 {angstrom} resolution. The structure reveals that the Aa1 VHH binds in the {alpha}-exosite of the BoNT/A Lc, far from the active site for catalysis. The study validates the utility of non-immune llama VHH libraries as a source of enzyme inhibitors and identifies the BoNT/A Lc {alpha}-exosite as a target for inhibitor development.« less

Sarcocystis masoni, n. sp. (Apicomplexa: Sarcocystidae), and redescription of Sarcocystis aucheniae from llama (Lama glama), guanaco (Lama guanicoe) and alpaca (Vicugna pacos)

USDA-ARS?s Scientific Manuscript database

There is considerable confusion concerning the species of Sarcocystis in South American camelids (SAC). Several species names have been used, however, proper descriptions are lacking. In the present paper we redescribe the macroscopic sarcocyst forming Sarcocystis aucheniae and describe and name the...

A new extant species of Electribius Crowson from Honduras (Coleoptera: Elateroidea: Artematopodidae).

PubMed

Gimmel, Matthew L; Bocakova, Milada

2015-03-05

A new extant species of Electribius Crowson, E. llamae sp.n., is described from Cusuco National Park, Cortés Province, Honduras. This new species lacks one of the supposed autapomorphies of the genus; therefore the definition of the genus requires modification. A revised key to the known extant species is presented, and their known distributions are mapped.

16 CFR 300.8 - Use of fiber trademark and generic names.

Code of Federal Regulations, 2011 CFR

2011-01-01

... is not the case. (g) The term fur fiber may be used to describe the hair or fur fiber or mixtures..., llama and vicuna. If the name, symbol, or depiction of any animal producing the hair or fur fiber is..., the percentage by weight of such hair or fur fiber in the total fiber weight of the wool product shall...

16 CFR 300.8 - Use of fiber trademark and generic names.

Code of Federal Regulations, 2010 CFR

2010-01-01

... is not the case. (g) The term fur fiber may be used to describe the hair or fur fiber or mixtures..., llama and vicuna. If the name, symbol, or depiction of any animal producing the hair or fur fiber is..., the percentage by weight of such hair or fur fiber in the total fiber weight of the wool product shall...

16 CFR 300.8 - Use of fiber trademark and generic names.

Code of Federal Regulations, 2012 CFR

2012-01-01

... is not the case. (g) The term fur fiber may be used to describe the hair or fur fiber or mixtures..., llama and vicuna. If the name, symbol, or depiction of any animal producing the hair or fur fiber is..., the percentage by weight of such hair or fur fiber in the total fiber weight of the wool product shall...

16 CFR 300.8 - Use of fiber trademark and generic names.

Code of Federal Regulations, 2014 CFR

2014-01-01

... is not the case. (g) The term fur fiber may be used to describe the hair or fur fiber or mixtures..., llama and vicuna. If the name, symbol, or depiction of any animal producing the hair or fur fiber is..., the percentage by weight of such hair or fur fiber in the total fiber weight of the wool product shall...

16 CFR 300.8 - Use of fiber trademark and generic names.

Code of Federal Regulations, 2013 CFR

2013-01-01

... is not the case. (g) The term fur fiber may be used to describe the hair or fur fiber or mixtures..., llama and vicuna. If the name, symbol, or depiction of any animal producing the hair or fur fiber is..., the percentage by weight of such hair or fur fiber in the total fiber weight of the wool product shall...

Effects of Sound, Vocabulary, and Grammar Learning Aptitude on Adult Second Language Speech Attainment in Foreign Language Classrooms

ERIC Educational Resources Information Center

Saito, Kazuya

2017-01-01

This study examines the relationship between different types of language learning aptitude (measured via the LLAMA test) and adult second language (L2) learners' attainment in speech production in English-as-a-foreign-language (EFL) classrooms. Picture descriptions elicited from 50 Japanese EFL learners from varied proficiency levels were analyzed…

Stabilization of an immunoglobulin fold domain by an engineered disulfide bond at the buried hydrophobic region.

PubMed

Hagihara, Yoshihisa; Mine, Shouhei; Uegaki, Koichi

2007-12-14

We report for the first time the stabilization of an immunoglobulin fold domain by an engineered disulfide bond. In the llama single-domain antibody, which has human chorionic gonadotropin as its specific antigen, Ala49 and Ile70 are buried in the structure. A mutant with an artificial disulfide bond at this position showed a 10 degrees C higher midpoint temperature of thermal unfolding than that without the extra disulfide bond. The modified domains exhibited an antigen binding affinity comparable with that of the wild-type domain. Ala49 and Ile70 are conserved in camel and llama single-domain antibody frameworks. Therefore, domains against different antigens are expected to be stabilized by the engineered disulfide bond examined here. In addition to the effect of the loop constraints in the unfolded state, thermodynamic analysis indicated that internal interaction and hydration also control the stability of domains with disulfide bonds. The change in physical properties resulting from mutation often causes unpredictable and destabilizing effects on these interactions. The introduction of a hydrophobic cystine into the hydrophobic region maintains the hydrophobicity of the protein and is expected to minimize the unfavorable mutational effects.

Bicarbonate secretion and solute absorption in forestomach of the llama.

PubMed

Rübsamen, K; Engelhardt, W V

1978-07-01

Bicarbonate appearance in the lumen and its relationship to solute absorption were studied in a Pavlov pouch in the cardiac region of the first compartment of the llama forestomach. HCO3- appearance showed no diurnal variation. HCO3- accumulation was highly dependent on the pH of the solution used. The HCO3- ion probably is formed from CO2 diffusing into the lumen from the serosal side, as a result of cell metabolism and of OH- ions. HCO3- accumulation was closely related to volatile fatty acid (VFA) absorption. The ratio of HCO3- appearance to VFA absorption depended on the pH of the solution. At a pH of 6.6, about 0.1 mol HCO3- and, at a pH of 7.8, 0.9 mol HCO3- appeared per mole absorbed VFA, indicating that at slightly alkaline pH nearly all H+ ions required for the nonionic absorption of VFA appeared to be delivered from the dissociation of H2CO3. Bicarbonate gain and VFA absorption were increased when animals were not fed for 48 h. Sodium absorption was related to VFA as well as water absorption.

Bluetongue disease and seroprevalence in South American camelids from the northwestern region of the United States.

PubMed

Allen, Andrew J; Stanton, James B; Evermann, James F; Fry, Lindsay M; Ackerman, Melissa G; Barrington, George M

2015-03-01

In late summer/early fall of 2013, 2 South American camelids from central Washington were diagnosed with fatal bluetongue viral disease, an event which is rarely reported. A 9-year-old intact male llama (Lama glama), with a 1-day history of anorexia, recumbency, and dyspnea before death. Abundant foam discharged from the mouth and nostrils, and the lungs were severely edematous on postmortem examination. Histologically, there was abundant intra-alveolar edema with fibrin. Hemorrhage and edema disrupted several other organs. Bluetongue viral RNA was detected by reverse transcription polymerase chain reaction (RT-PCR), and serotype 11 was identified by sequencing a segment of the VP2 outer capsid gene. Approximately 1 month later, at a site 150 miles north of the index case, a 2-year-old female alpaca with similar, acutely progressive clinical signs was reported. A postmortem examination was performed, and histologic lesions from the alpaca were similar to those of the llama, and again serotype 11 was detected by PCR. The occurrence of bluetongue viral infection and disease is described in the context of seasonal Bluetongue virus activity within the northwestern United States and southwestern Canada. © 2015 The Author(s).

[The occurrence of "Candidatus Mycoplasma haemolamae" infections in clinically asymptomatic South American Camelids in Austria].

PubMed

Franz, Sonja; Spergser, Joachim; Schwendenwein, Ilse; Stanitznig, Anna; Lambacher, Bianca; Tichy, Alexander; Wittek, Thomas

2016-01-01

Reports of CMhl infections in South American Camelids in Europe are only available from the United Kingdom and Switzerland. Knowing that CMhl infections can lead to severe disease resulting in death if combined with other diseases or stress, it was the aim of this study to assess prevalence data from camelids in Austria. In comparison to the previous studies a representative number of camelids was investigated nationwide. Data were assessed due to differences in geographical region, age, sex, species, and origin. A relatively high prevalence of 25.8% was recorded. CMhl was detected significantly more often in alpacas (Vicunja pacos) than in llamas (Lama glama) and more frequently in animals younger than 2 years. Additionally regional differences have been observed, which might be due to climatic differences and/or variations in insect vectors. In this study apperantly clinical healthy animals were shown to be infected with CMhl. Camelids infected with CMhl are a pathogen reservoir. The results of this study indicate different risk levels of infection between llamas and alpacas and between younger and older animals. The data presented underline the necessity of further studies on CMhlI infections in South American Camelids.

Isolation of Llama Antibody Fragments for Prevention of Dandruff by Phage Display in Shampoo

PubMed Central

Dolk, Edward; van der Vaart, Marcel; Lutje Hulsik, David; Vriend, Gert; de Haard, Hans; Spinelli, Silvia; Cambillau, Christian; Frenken, Leon; Verrips, Theo

2005-01-01

As part of research exploring the feasibility of using antibody fragments to inhibit the growth of organisms implicated in dandruff, we isolated antibody fragments that bind to a cell surface protein of Malassezia furfur in the presence of shampoo. We found that phage display of llama single-domain antibody fragments (VHHs) can be extended to very harsh conditions, such as the presence of shampoo containing nonionic and anionic surfactants. We selected several VHHs that bind to the cell wall protein Malf1 of M. furfur, a fungus implicated in causing dandruff. In addition to high stability in the presence of shampoo, these VHHs are also stable under other denaturing conditions, such as high urea concentrations. Many of the stable VHHs were found to contain arginine at position 44. Replacement of the native amino acid at position 44 with arginine in the most stable VHH that lacked this arginine resulted in a dramatic further increase in the stability. The combination of the unique properties of VHHs together with applied phage display and protein engineering is a powerful method for obtaining highly stable VHHs that can be used in a wide range of applications. PMID:15640220

SdAb heterodimer formation using leucine zippers

NASA Astrophysics Data System (ADS)

Goldman, Ellen R.; Anderson, George P.; Brozozog-Lee, P. Audrey; Zabetakis, Dan

2013-05-01

Single domain antibodies (sdAb) are variable domains cloned from camel, llama, or shark heavy chain only antibodies, and are among the smallest known naturally derived antigen binding fragments. SdAb derived from immunized llamas are able to bind antigens with high affinity, and most are capable of refolding after heat or chemical denaturation to bind antigen again. We hypothesized that the ability to produce heterodimeric sdAb would enable reagents with the robust characteristics of component sdAb, but with dramatically improved overall affinity through increased avidity. Previously we had constructed multimeric sdAb by genetically linking sdAb that bind non-overlapping epitopes on the toxin, ricin. In this work we explored a more flexible approach; the construction of multivalent binding reagents using multimerization domains. We expressed anti-ricin sdAb that recognize different epitopes on the toxin as fusions with differently charged leucine zippers. When the initially produced homodimers are mixed the leucine zipper domains will pair to produce heterodimers. We used fluorescence resonance energy transfer to confirm heterodimer formation. Surface plasmon resonance, circular dichroism, enzyme linked immunosorbent assays, and fluid array assays were used to characterize the multimer constructs, and evaluate their utility in toxin detection.

Surgical repair of femoral fractures in New World camelids: five cases (1996-2003).

PubMed

Shoemaker, R W; Wilson, D G

2007-04-01

Five New World camelids were admitted to the Western College of Veterinary Medicine between 1996 and 2003 for evaluation of femoral fractures. There were three alpacas and two llamas. Four of the animals were female and three were less than 3 months of age. Fracture configurations consisted of distal physeal fractures (three), a comminuted diaphyseal/metaphyseal fracture, and a transverse diaphyseal fracture. Fractures were diagnosed with a combination of physical examination and radiographs in all cases. All five fractures were repaired with internal fixation and three animals were discharged from the hospital with fractures that healed. One cria underwent successful internal fixation but died from pulmonary oedema during recovery from anaesthesia. Postoperative complications were rare and limited to inadequate fracture stability in one alpaca and prolonged recovery to weight bearing in another. One llama with a comminuted metaphyseal fracture, repaired with a 4.5 mm dynamic compression plate, subsequently had catastrophic failure of the bone 17 days after surgery. Overall the clients were pleased with the outcome of discharged animals. Although femoral fractures are considered rare, they pose a unique opportunity for the large animal veterinarian to successfully achieve fracture union with the aid of internal fixation.

Blood typing South American camelids.

PubMed

Miller, W J; Hollander, P J; Franklin, W L

1985-01-01

Preliminary blood typing tests were made on New World camelids, guanacos, llamas, and two hybrids. Erythrocyte samples were tested against a battery of cattle blood typing reagents. Three different reagents were prepared from rabbit anti-erythrocyte sera. Transferrin variation and lectin polymorphism also were observed. No naturally occurring isoantibodies were found. Blood typing tests of New World camelids were shown to be feasible for studies of taxonomic relationships.

First cloned Bactrian camel (Camelus bactrianus) calf produced by interspecies somatic cell nuclear transfer: A step towards preserving the critically endangered wild Bactrian camels.

PubMed

Wani, Nisar Ahmad; Vettical, Binoy S; Hong, Seung B

2017-01-01

Studies were conducted to explore the possibility of employing dromedary camel (Camelus dromedarius) oocytes as recipient cytoplasts for the development of interspecies somatic cell nuclear transfer (iSCNT) embryos using skin fibroblast cells of an adult Bactrian camel (Camelus bactrianus) and Llama (Llama glama) as donor nuclei. Also, the embryos reconstructed with Bactrian cells were transferred into the uterus of synchronized dromedary camel recipients to explore the possibility of using them as surrogate mothers. Serum-starved skin fibroblast cells were injected into the perivitelline space of enucleated mature oocytes, collected from super-stimulated dromedary camels, and fused using an Eppendorf electroporator. After activation with 5μM ionomycin and 6-dimethylaminopurine, they were cultured at 38.5°C in an atmosphere of 5% CO2, 5% O2, and 90% N2 in air. In experiment 1, Day 7 blastocysts were stained with Hoechst to count their cell numbers, while in experiment 2, they were transferred to synchronized dromedary recipients. A lower number (P < 0.05) of blastocysts were obtained from reconstructs utilizing fibroblast cells from Llama when compared with those reconstructed with dromedary and Bactrian fibroblast cells. However, no difference was observed in their cell numbers. In experiment 2, a higher (P < 0.05) proportion of blastocysts were obtained from the cleaved embryos reconstructed with Bactrian fibroblast cells when compared to those reconstructed with dromedary cells. Twenty-six Day 7 blastocysts reconstructed with Bactrian cells were transferred to 23 synchronized dromedary recipients with 5 pregnancies established on Day 30, however, only one of the pregnancies developed to term and a healthy calf weighing 33 kgs was born after completing 392 days of gestation. Unfortunately, the calf died on day 7 due to acute septicemia. In conclusion, the present study reports, for the first time, birth of a cloned Bactrian calf by iSCNT using dromedary camel as a source for oocytes as well as a surrogate for carrying the pregnancy to term.

First cloned Bactrian camel (Camelus bactrianus) calf produced by interspecies somatic cell nuclear transfer: A step towards preserving the critically endangered wild Bactrian camels

PubMed Central

Vettical, Binoy S.; Hong, Seung B.

2017-01-01

Studies were conducted to explore the possibility of employing dromedary camel (Camelus dromedarius) oocytes as recipient cytoplasts for the development of interspecies somatic cell nuclear transfer (iSCNT) embryos using skin fibroblast cells of an adult Bactrian camel (Camelus bactrianus) and Llama (Llama glama) as donor nuclei. Also, the embryos reconstructed with Bactrian cells were transferred into the uterus of synchronized dromedary camel recipients to explore the possibility of using them as surrogate mothers. Serum-starved skin fibroblast cells were injected into the perivitelline space of enucleated mature oocytes, collected from super-stimulated dromedary camels, and fused using an Eppendorf electroporator. After activation with 5μM ionomycin and 6-dimethylaminopurine, they were cultured at 38.5°C in an atmosphere of 5% CO2, 5% O2, and 90% N2 in air. In experiment 1, Day 7 blastocysts were stained with Hoechst to count their cell numbers, while in experiment 2, they were transferred to synchronized dromedary recipients. A lower number (P < 0.05) of blastocysts were obtained from reconstructs utilizing fibroblast cells from Llama when compared with those reconstructed with dromedary and Bactrian fibroblast cells. However, no difference was observed in their cell numbers. In experiment 2, a higher (P < 0.05) proportion of blastocysts were obtained from the cleaved embryos reconstructed with Bactrian fibroblast cells when compared to those reconstructed with dromedary cells. Twenty-six Day 7 blastocysts reconstructed with Bactrian cells were transferred to 23 synchronized dromedary recipients with 5 pregnancies established on Day 30, however, only one of the pregnancies developed to term and a healthy calf weighing 33 kgs was born after completing 392 days of gestation. Unfortunately, the calf died on day 7 due to acute septicemia. In conclusion, the present study reports, for the first time, birth of a cloned Bactrian calf by iSCNT using dromedary camel as a source for oocytes as well as a surrogate for carrying the pregnancy to term. PMID:28545049

An Assessment of ELINT Exploitation for Situational Awareness Visualisations on Operator Situational Awareness

DTIC Science & Technology

2006-10-01

C L A S I F I C A T I O N An Assessment of ELINT Exploitation for Situational Awareness Visualisations on Operator Situational...environment. The An Assessment of ELINT Exploitation for Situational Awareness Visualisations on Operator Situational Awareness EXECUTIVE SUMMARY...1. ELEXSA’s Process of Sequential Enrichment. 2 Figure 2. Sample ELEXSA Visualisation . 3 Figure 3. Example Llama/Cheetah Network Layout. 5

Reproduction in female South American domestic camelids.

PubMed

Sumar, J B

1999-01-01

Alpacas and llamas are induced ovulators. They show marked reproductive seasonality in the Andean region, but under Northern Hemisphere conditions of feeding and management, they are non-seasonal breeders. Puberty is attained when they reach 50% of adult body weight. When they are not exposed to a male, females show successive waves of follicular maturation and atresia. Growth, maintenance and regression of a follicle each require an average of 4 and 6 days in alpacas and llamas, respectively. After sterile mating, progesterone concentrations in blood were increased from day 5, reached maximum concentrations on day 7-8, and declined rapidly at 9-10 days after mating. A fertile mating results in formation of a corpus luteum that remains functional throughout gestation. The duration of gestation is 340-346 days. Almost all fetuses were found to occupy the left uterine horn, even though ovulation occurs from both ovaries with equal frequency. Several methods of pregnancy diagnosis have been described. Mating is recommended within 15-20 days after parturition to obtain good fertility rates and one offspring per year. The factors that contribute to high rates of embryonic mortality are unknown. Reproductive technologies, such as AI, superovulation, embryo transfer and IVF, have not been used very extensively in these species but can be successfully applied.

Comparison of two cooling protocols for llama semen: with and without collagenase and seminal plasma in the medium.

PubMed

Carretero, M I; Giuliano, S M; Arraztoa, C C; Santa Cruz, R C; Fumuso, F G; Neild, D M

2017-08-01

Seminal plasma (SP) of South American Camelids could interfere with the interaction of spermatozoa with the extenders; therefore it becomes necessary to improve semen management using enzymatic treatment. Our objective was to compare two cooling protocols for llama semen. Twelve ejaculates were incubated in 0.1% collagenase and then were divided into two aliquots. One was extended in lactose and egg yolk (LEY) (Protocol A: collagenase and SP present). The other aliquot was centrifuged, and the pellet was resuspended in LEY (Protocol B: collagenase and SP absent). Both samples were maintained at 5°C during 24 hr. Routine and DNA evaluations were carried out in raw and cooled semen. Both cooling protocols maintained sperm viability, membrane function and DNA fragmentation, with Protocol A showing a significantly lowered total and progressive motility (p < .05) and Protocol B showing a significant increase in chromatin decondensation (p < .05). Protocol A avoids centrifugation, reducing processing times and making application in the field simpler. However, as neither protocol showed a significant superiority over the other, studies should be carried out in vivo to evaluate the effect on pregnancy rates of the presence of collagenase and SP in semen samples prior to either cooling or freeze-thawing. © 2016 Blackwell Verlag GmbH.

Prevalence of Mycobacterium avium subspecies paratuberculosis and hepatitis E in New World camelids in Austria.

PubMed

Stanitznig, A; Khol, J L; Lambacher, B; Franz, S; Wittek, T; Kralik, P; Slana, I; Vasickova, P

2017-07-07

Mycobacterium avium subspecies paratuberculosis (MAP) is the causative agent of paratuberculosis in domestic ruminants and New World Camelids (NWC). Hepatitis E virus (HEV) is an important public health concern worldwide. The virus has been identified in several species, some of them serving as a reservoir for zoonotic HEV strains. Husbandry and breeding of llamas and alpacas have increased in Austria in recent years. Therefore, the aim of the present study was to evaluate the prevalence of MAP and HEV in NWC in Austria. Altogether 445 animals, originating from 78 farms were enrolled in the study. Of the animals sampled, 184 (41.35%) were llamas and 261 (58.65%) were alpacas. 443 blood samples for MAP-ELISA and 399 faecal samples for quantitative PCR (qPCR) and culture for MAP as well as for HEV detection by RT-qPCR have been collected. All of the 399 animals tested for shedding of MAP were negative by faecal solid culture. Using qPCR, 15 (3.8%) of the animals were MAP positive and 384 (96.2%) negative. Out of the 443 serum samples examined for specific antibodies against MAP by ELISA, 6 (1.4%) were positive, 1 (0.2%) was questionable and 436 (98.4%) samples were negative. All faecal samples were tested negative for HEV.

Antibody response to the epsilon toxin of Clostridium perfringens following vaccination of Lama glama crias.

PubMed

Bentancor, Adriana B; Halperin, Pablo; Flores, Myriam; Iribarren, Fabián

2009-09-15

Enterotoxaemia produced by Clostridium perfringens A, C and D is an important cause of mortality in young llamas. There is no data on antibody responses following vaccination with epsilon toxin. Twenty-six L. glama crias were divided into four groups which were vaccinated with a commercial vaccine (Mancha Gangrena Enterotoxemia, Instituto Rosembusch Sociedad Anónima, Argentina) on days 0, 21 and 42 or left as unvaccinated controls. An indirect ELISA was compared with the mouse neutralization test (MNT) for measuring titers to C. perfringens type D epsilon toxin and used to determine titers in sera taken before vaccination and 16, 28, 49, 59, and 93 days later. The ELISA gave comparable results to the MNT and showed animals vaccinated once failed to develop raised titers. A week following a second vaccination, mean antibody titers rose significantly (P < 0.05) and 7/12 animals developed high titers which were present in only one animal at the end of the study (day 93). A third vaccination resulted in a decrease in mean antibody titers a week later. Llamas develop antibodies to Clostridium perfringens type D epsilon toxin after two vaccinations at a 21-day interval. Further studies are indicated to determine if these inoculations protect against enterotoxemia and the most appropriate vaccination schedule.

Clinical findings and survival in 56 sick neonatal New World camelids.

PubMed

Bertin, F R; Squires, J M; Kritchevsky, J E; Taylor, S D

2015-01-01

Information pertaining to clinical presentation and outcome of neonatal New World camelids (NWC) is limited when compared to calves and foals. Values of variables at admission and subsequent treatment would predict survival in sick neonatal NWC. Fifty-six client-owned sick neonatal NWC presented over a 10-year period to the Purdue University Veterinary Teaching Hospital. A retrospective study was performed. Inclusion criteria were NWC less than 30 days of age with complete medical records that presented between 2000 and 2010. The median age at presentation was 1 day (range 1-20). The most common diagnoses were systemic inflammatory response syndrome (50%), congenital defects (41%), ophthalmic lesions (21%), sepsis (16%), and gastrointestinal diseases (16%). Sixty-six percent of NWC survived to discharge. Clinicopathologic findings on admission were variable and not specific for disorders. Factors associated with survival were absence of choanal atresia (P = .001, OR: 55.9 [2.5-1,232]), administration of llama plasma (P = .013, OR: 4.9 [1.4-17.7]), and antimicrobial treatment with trimethoprim-sulfamethoxazole (TMS) (P = .016, OR: 6.5 [1.3-32.2]). The use of antibiotics, particularly TMS, and llama plasma are recommended in sick neonatal NWC. Results from this study could contribute toward defining a NWC-specific sepsis scoring system. Copyright © 2014 by the American College of Veterinary Internal Medicine.

Anaerobic co-digestion of aquatic flora and quinoa with manures from Bolivian Altiplano.

PubMed

Alvarez, René; Lidén, Gunnar

2008-01-01

Quinoa stalk (Chenopodium quinoa Willd.) from agricultural crop residue, totora (Schoenoplectus tatora) and o-macrophytes (aquatic flora) from Lake Titicaca (on the Bolivian Altiplano) were studied in a wet anaerobic co-digestion process together with manure from llama, cow and sheep. Anaerobic semi-continuous experiments were performed in (10) 2-l reactors at a temperature of 25 degrees C with 30 days of hydraulic retention time (HRT) and an organic loading rate (OLR) of 1.8 kg VS m(-3) d(-1). Totora was found to be the best co-substrate. In mixture ratios of 1:1 (VS basis), it increased the biogas productivity by 130% for llama manure, 60% for cow manure, and 40% for sheep manure. It was possible to use up to 58% (VS basis) of totora in the substrate. Higher concentrations (including pure totora) could not be digested, as that caused acidification problems similar to those caused by other lignocellulosic materials. When quinoa and o-macrophytes were used as co-substrates, the increase in biogas productivity was slightly less. However, these co-substrates did not cause any operational problems. An additional advantage of quinoa and o-macrophytes was that they could be used in any proportion (even in pure form) without causing any destabilization problems in the anaerobic digestion process.

Single-domain antibodies that compete with the natural ligand fibroblast growth factor block the internalization of the fibroblast growth factor receptor 1

DOE Office of Scientific and Technical Information (OSTI.GOV)

Veggiani, Gianluca; Ossolengo, Giuseppe; Aliprandi, Marisa

2011-05-20

Highlights: {yields} Recombinant antibodies for FGFR1 were isolated from a llama naive library in VHH format. {yields} These antibodies compete with the natural ligand FGF-2 for the same epitope on FGFR1. {yields} The antibody competition inhibits the FGF-2-dependent internalization of FGFR1. -- Abstract: Single-domain antibodies in VHH format specific for fibroblast growth factor receptor 1 (FGFR1) were isolated from a phage-display llama naive library. In particular, phage elution in the presence of the natural receptor ligand fibroblast growth factor (FGF) allowed for the identification of recombinant antibodies that compete with FGF for the same region on the receptor surface. Thesemore » antibodies posses a relatively low affinity for FGFR1 and were never identified when unspecific elution conditions favoring highly affine binders were applied to panning procedures. Two populations of competitive antibodies were identified that labeled specifically the receptor-expressing cells in immunofluorescence and recognize distinct epitopes. Antibodies from both populations effectively prevented FGF-dependent internalization and nuclear accumulation of the receptor in cultured cells. This achievement indicates that these antibodies have a capacity to modulate the receptor physiology and, therefore, constitute powerful reagents for basic research and a potential lead for therapeutic applications.« less

Genome sequence of "Candidatus Mycoplasma haemolamae" strain purdue, a red blood cell pathogen of alpacas (Vicugna pacos) and llamas (Lama glama).

PubMed

Guimaraes, Ana M S; Toth, Balazs; Santos, Andrea P; do Nascimento, Naíla C; Kritchevsky, Janice E; Messick, Joanne B

2012-11-01

We report the complete genome sequence of "Candidatus Mycoplasma haemolamae," an endemic red-cell pathogen of camelids. The single, circular chromosome has 756,845 bp, a 39.3% G+C content, and 925 coding sequences (CDSs). A great proportion (49.1%) of these CDSs are organized into paralogous gene families, which can now be further explored with regard to antigenic variation.

9 CFR 130.2 - User fees for individual animals and certain birds quarantined in the APHIS-owned or -operated...

Code of Federal Regulations, 2014 CFR

2014-01-01

... grams 18.00 19.00 19.00 20.00 21.00 Domestic or zoo animals (except equines, birds, and poultry): Bison, bulls, camels, cattle, or zoo animals 144.00 149.00 153.00 158.00 162.00 All others, including, but not limited to, alpacas, llamas, goats, sheep, and swine 38.00 39.00 40.00 42.00 43.00 Equines (including zoo...

9 CFR 130.2 - User fees for individual animals and certain birds quarantined in the APHIS-owned or -operated...

Code of Federal Regulations, 2013 CFR

2013-01-01

... grams 18.00 19.00 19.00 20.00 21.00 Domestic or zoo animals (except equines, birds, and poultry): Bison, bulls, camels, cattle, or zoo animals 144.00 149.00 153.00 158.00 162.00 All others, including, but not limited to, alpacas, llamas, goats, sheep, and swine 38.00 39.00 40.00 42.00 43.00 Equines (including zoo...

9 CFR 130.2 - User fees for individual animals and certain birds quarantined in the APHIS-owned or -operated...

Code of Federal Regulations, 2012 CFR

2012-01-01

... grams 18.00 19.00 19.00 20.00 21.00 Domestic or zoo animals (except equines, birds, and poultry): Bison, bulls, camels, cattle, or zoo animals 144.00 149.00 153.00 158.00 162.00 All others, including, but not limited to, alpacas, llamas, goats, sheep, and swine 38.00 39.00 40.00 42.00 43.00 Equines (including zoo...

Dead space and tidal volume of the giraffe compared with some other mammals.

PubMed

Hugh-Jones, P; Barter, C E; Hime, J M; Rusbridge, M M

1978-10-01

The ventilation, tidal volume and anatomical dead-space were measured in a living giraffe and compared with similar measurements in a camel, red deer, llama and man. The giraffe had a resting tidal volume of about 3.3 litres with a dead-space/tidal-volume ratio of 0.34. The giraffe breathes slowly, apparently because of the unusually small diameter of its trachea relative to its length, compared with known measurement in other mammals.

A review of the nutritional content and technological parameters of indigenous sources of meat in South America.

PubMed

Saadoun, A; Cabrera, M C

2008-11-01

Meat yields, proximate compositions, fatty acids compositions and technological parameters are reviewed for species which might be further developed as indigenous sources of meat in South America. These include the alpaca (Lama pacos), capybara (Hydrochoerus hydrochaeris), guanaco (Lama guanicoe), llama (Lama glama), nutria (Myocastor coypus), collared peccary (Tayassu tajacu), greater rhea (Rhea americana), lesser rhea (Rhea pennata), yacare (Caiman crocodilus yacare), tegu lizard (Tupinambis merianae) and green iguana (Iguana iguana).

Genome Sequence of “Candidatus Mycoplasma haemolamae” Strain Purdue, a Red Blood Cell Pathogen of Alpacas (Vicugna pacos) and Llamas (Lama glama)

PubMed Central

Toth, Balazs; Santos, Andrea P.; do Nascimento, Naíla C.; Kritchevsky, Janice E.

2012-01-01

We report the complete genome sequence of “Candidatus Mycoplasma haemolamae,” an endemic red-cell pathogen of camelids. The single, circular chromosome has 756,845 bp, a 39.3% G+C content, and 925 coding sequences (CDSs). A great proportion (49.1%) of these CDSs are organized into paralogous gene families, which can now be further explored with regard to antigenic variation. PMID:23105057

[Unilateral laparoscopic cryptorchidectomy in a llama].

PubMed

Zulauf, M; Bergadano, A; Steiner, A

2002-05-01

Cryptorchism is an inherited congenital abnormality characterized by the failure of one or both testes to descend into the scrotum. Diagnosis can be difficult if the retained testicular tissue cannot be localized during clinical examination and the history of the animal is unknown, or if the animal shows persistent male behaviour despite the history of successful previous castration. The present case report describes the history of a cryptorchid Lama and the techniques of general anesthesia and laparoscopic cryptorchidectomy advocated.

Camelid cardiology.

PubMed

Margiocco, Marco L; Scansen, Brian A; Bonagura, John D

2009-07-01

Cardiovascular disorders, although not thoroughly described in the literature, are frequently diagnosed in South American camelids, causing morbidity, mortality, and loss of production. Definitive confirmation concerning the heritability of cardiac defects in these species is lacking; however, this potential exists and should be taken into account when counseling breeders and owners. This article describes the diagnosis and treatment of cardiovascular diseases in llamas and alpacas and reviews the most recent literature. Unique aspects of the cardiovascular physiology in these species are also reviewed.

Anaerobic co-digestion of aquatic flora and quinoa with manures from Bolivian Altiplano

DOE Office of Scientific and Technical Information (OSTI.GOV)

Alvarez, Rene; Liden, Gunnar

2008-07-01

Quinoa stalk (Chenopodium quinoa Willd.) from agricultural crop residue, totora (Schoenoplectus tatora) and o-macrophytes (aquatic flora) from Lake Titicaca (on the Bolivian Altiplano) were studied in a wet anaerobic co-digestion process together with manure from llama, cow and sheep. Anaerobic semi-continuous experiments were performed in (10) 2-l reactors at a temperature of 25 deg. C with 30 days of hydraulic retention time (HRT) and an organic loading rate (OLR) of 1.8 kg VS m{sup -3} d{sup -1}. Totora was found to be the best co-substrate. In mixture ratios of 1:1 (VS basis), it increased the biogas productivity by 130% formore » llama manure, 60% for cow manure, and 40% for sheep manure. It was possible to use up to 58% (VS basis) of totora in the substrate. Higher concentrations (including pure totora) could not be digested, as that caused acidification problems similar to those caused by other lignocellulosic materials. When quinoa and o-macrophytes were used as co-substrates, the increase in biogas productivity was slightly less. However, these co-substrates did not cause any operational problems. An additional advantage of quinoa and o-macrophytes was that they could be used in any proportion (even in pure form) without causing any destabilization problems in the anaerobic digestion process.« less

Evaluation of Lama glama semen viscosity with a cone-plate rotational viscometer.

PubMed

Casaretto, C; Martínez Sarrasague, M; Giuliano, S; Rubin de Celis, E; Gambarotta, M; Carretero, I; Miragaya, M

2012-05-01

Llama semen is highly viscous. This characteristic is usually evaluated subjectively by measuring the thread formed when carefully pippeting a sample of semen. The aims of this study were (i) to objectively determine and analyse llama semen viscosity, (ii) to compare semen viscosity between ejaculates of the same male as well as between different males, (iii) to study the correlation between viscosity and other semen characteristics and (iv) to evaluate the effect of collagenase on semen viscosity. Semen viscosity was evaluated using a cone-plate Brookfield rotational viscometer. A non Newtonian, pseudoplastic behaviour was observed in the 45 semen samples evaluated. Rheological parameters were determined obtaining the following results (mean ± SD): apparent viscosity at 11.5 s(-1): 46.71 ± 26.8 cpoise and at 115 s(-1): 12.61 ± 4.1 cpoise; structural viscosity (K) (dyne s cm(-2)): 2.18 ± 1.4 and coefficient of consistency (n): 0.45 ± 0.1. Statistical differences were found between different ejaculates of the same male for structural viscosity and apparent viscosity at 11.5 s(-1) (P < 0.01). Correlation was found only between coefficient of consistency (n) and sperm concentration (P < 0.01). Significant differences for coefficient of consistency (n) and viscosity at 115 s(-1) were found between samples incubated with and without collagenase (P < 0.05). © 2011 Blackwell Verlag GmbH.

Prairie rattlesnake envenomation in 27 New World camelids.

PubMed

Sonis, J M; Hackett, E S; Callan, R J; Holt, T N; Hackett, T B

2013-01-01

Morbidity and case fatality from rattlesnake envenomation is regionally specific because of variability in relative toxicity of the species of snake encountered. A previous report of rattlesnake envenomation in New World camelids (NWC) from the western coastal United States documented high case fatality rates and guarded prognosis for survival. To describe clinical findings, treatments, and outcome of NWC with prairie rattlesnake (Crotalus viridis viridis) envenomation in the Rocky Mountain region of the United States. Twenty-seven NWC admitted to the Colorado State University Veterinary Teaching Hospital for evaluation of acute rattlesnake envenomation between 1992 and 2012. Medical records of NWC evaluated for rattlesnake envenomation as coded by the attending clinician and identified by a database search were reviewed retrospectively. Month of admission, signalment, area of bite, clinical and clinicopathologic data, treatments, and outcome were recorded. Twenty-five llamas and 2 alpacas were admitted for envenomation. Llamas were overrepresented compared to hospital caseload. The face was the most common site of envenomation, observed in 96% of recorded cases. Presenting clinical signs included fever, tachypnea, tachycardia, and respiratory distress. Nine animals required a tracheotomy. Median hospitalization time was 3 days and overall survival rate was 69%. Case fatality rate for prairie rattlesnake envenomation in NWC was lower than that reported in the Western coastal region of the United States and similar to that reported for prairie rattlesnake envenomation in horses. Copyright © 2013 by the American College of Veterinary Internal Medicine.

Crystal Structure of the Neutralizing Llama VHH D7 and Its Mode of HIV-1 gp120 Interaction

PubMed Central

Hinz, Andreas; Lutje Hulsik, David; Forsman, Anna; Koh, Willie Wee-Lee; Belrhali, Hassan; Gorlani, Andrea; de Haard, Hans; Weiss, Robin A.; Verrips, Theo; Weissenhorn, Winfried

2010-01-01

HIV-1 entry into host cells is mediated by the sequential binding of the envelope glycoprotein gp120 to CD4 and a chemokine receptor. Antibodies binding to epitopes overlapping the CD4-binding site on gp120 are potent inhibitors of HIV entry, such as the llama heavy chain antibody fragment VHH D7, which has cross-clade neutralizing properties and competes with CD4 and mAb b12 for high affinity binding to gp120. We report the crystal structure of the D7 VHH at 1.5 Å resolution, which reveals the molecular details of the complementarity determining regions (CDR) and substantial flexibility of CDR3 that could facilitate an induced fit interaction with gp120. Structural comparison of CDRs from other CD4 binding site antibodies suggests diverse modes of interaction. Mutational analysis identified CDR3 as a key component of gp120 interaction as determined by surface plasmon resonance. A decrease in affinity is directly coupled to the neutralization efficiency since mutations that decrease gp120 interaction increase the IC50 required for HIV-1 IIIB neutralization. Thus the structural study identifies the long CDR3 of D7 as the key determinant of interaction and HIV-1 neutralization. Furthermore, our data confirm that the structural plasticity of gp120 can accommodate multiple modes of antibody binding within the CD4 binding site. PMID:20463957

Identification and isolation of stimulator of interferon genes (STING): an innate immune sensory and adaptor gene from camelids.

PubMed

Premraj, A; Aleyas, A G; Nautiyal, B; Rasool, T J

2013-10-01

The mechanism by which type I interferon-mediated antiviral response is mounted by hosts against invading pathogen is an intriguing one. Of late, an endoplasmic reticulum transmembrane protein encoded by a gene called stimulator of interferon genes (STING) is implicated in the innate signalling pathways and has been identified and cloned in few mammalian species including human, mouse and pig. In this article, we report the identification of STING from three different species of a highly conserved family of mammals - the camelids. cDNAs encoding the STING of Old World camels - dromedary camel (Camelus dromedarius) and bactrian camel (Camelus bactrianus) and a New World camel - llama (Llama glama) were amplified using conserved primers and RACE. The complete STING cDNA of dromedary camel is 2171 bp long with a 706-bp 5' untranslated regions (UTR), an 1137-bp open reading frame (ORF) and a 328-bp 3' UTR. Sequence and phylogenetic analysis of the ORF of STING from these three camelids indicate high level of similarity among camelids and conservation of critical amino acid residues across different species. Quantitative real-time PCR analysis revealed high levels of STING mRNA expression in blood, spleen, lymph node and lung. The identification of camelid STING will help in better understanding of the role of this molecule in the innate immunity of the camelids and other mammals. © 2013 John Wiley & Sons Ltd.

Brightness of venous blood in South American camelids: implications for jugular catheterization.

PubMed

Grint, Nicola; Dugdale, Alexandra

2009-01-01

To compare the brightness of South American camelid venous blood to that of Equidae. Prospective clinical evaluation. Twelve South American camelids (eight llamas, four alpacas), eight horses and ponies (control group). Appropriately sized catheters were placed in the jugular vein of each animal under local anaesthesia. The blood spilt before the catheter was capped was caught on a white tile. A sample of blood was drawn for blood-gas analysis. The brightness of the blood (both on the tile and in the syringe) was matched to a colour chart (1 = darkest red, 8 = brightest red) by a single observer under bright light conditions. Packed cell volume (PCV) and partial pressure of oxygen (PvO(2)) in the blood were also measured on the syringe blood. Normally distributed data were compared using a two tailed t-test, and non-normally distributed data were compared using a Mann-Whitney U-test. Significance was set at p < 0.05. Camelid venous blood was significantly brighter red than that of horses and ponies both on the white tile (p = 0.0003) and in the syringe (p = 0.0001). PCV was significantly lower in camelids (32 +/- 4%) compared with horses (37 +/- 5%). Partial pressure of oxygen values were similar between groups. Jugular venous blood in alpacas and llamas is significantly brighter red than that of horses. Colour should not be used as a sole determinant of venous or arterial catheterization in this species.

The effect of ovarian follicle size on pituitary and ovarian responses to copulation in domesticated South American camelids.

PubMed

Bravo, P W; Stabenfeldt, G H; Lasley, B L; Fowler, M E

1991-10-01

The relation of ovarian follicle size to pituitary and ovarian responses to copulation was studied in domesticated South American camelids (llamas and alpacas). Females from each species were divided into four groups according to follicle size: small (4-5 mm), growing (6-7 mm), mature (8-12 mm), and regressing (10-7 mm). The pituitary response to copulation was determined by analysis of LH and FSH concentrations in plasma. The ovarian response to copulation was determined by ultrasonography and by analysis of estrone sulfate (follicular status) and pregnanediol glucuronide (luteal status) concentrations in urine. Females with small follicles (4-5 mm) released less LH after copulation than did those with larger follicles, and ovulation was not induced. Females with growing and mature follicles (7-12 mm) released LH in response to copulation that was adequate to induce ovulation and to initiate normal luteal activity. While copulation-induced LH release in females with regressing follicles was similar to that released in animals with growing and mature follicles, regressing follicles were luteinized instead of being ovulated. The luteal structure formed as a result of luteinization of follicles had a short life span, i.e., 5.1 days. Copulation-induced LH release was significantly higher in llamas vs. alpacas in animals with mature or regressing follicles, but not in those with small or growing follicles. Urinary estrone sulfate and pregnanediol glucuronide concentrations correlated positively with the presence of follicles and corpora lutea, respectively.

Llama heavy-chain antibody fragments efficiently remove toxic shock syndrome toxin 1 from plasma in vitro but not in experimental porcine septic shock.

PubMed

Brummelhuis, Walter J; Joles, Jaap A; Stam, Jord C; Adams, Hendrik; Goldschmeding, Roel; Detmers, Frank J; El Khattabi, Mohamed; Maassen, Bram T; Verrips, C Theo; Braam, Branko

2010-08-01

Staphylococcus aureus produces the superantigen toxic shock syndrome toxin 1 (TSST-1). When the bacterium invades the human circulation, this toxin can induce life-threatening gram-positive sepsis. Current sepsis treatment does not remove bacterial toxins. Variable domains of llama heavy-chain antibodies (VHH) against toxic shock syndrome toxin 1 ([alpha]-TSST-1 VHH) were previously found to be effective in vitro. We hypothesized that removing TSST-1 with [alpha]-TSST-1 VHH hemofiltration filters would ameliorate experimental sepsis in pigs. After assessing in vitro whether timely removing TSST-1 interrupted TSST-1-induced mononuclear cell TNF-[alpha] production, VHH-coated filters were applied in a porcine sepsis model. Clinical course, survival, plasma interferon [gamma], and TSST-1 levels were similar with and without VHH-coated filters as were TSST-1 concentrations before and after the VHH filter. Plasma TSST-1 levels were much lower than anticipated from the distribution of the amount of infused TSST-1, suggesting compartmentalization to space or adhesion to surface not accessible to hemofiltration or pheresis techniques. Removing TSST-1 from plasma was feasible in vitro. However, the [alpha]-TSST-1 VHH adsorption filter-based technique was ineffective in vivo, indicating that improvement of VHH-based hemofiltration is required. Sequestration likely prevented the adequate removal of TSST-1. The latter warrants further investigation of TSST-1 distribution and clearance in vivo.

Esophageal dysfunction in four alpaca crias and a llama cria with vascular ring anomalies.

PubMed

McKenzie, Erica C; Seguin, Bernard; Cebra, Christopher K; Margiocco, Marco L; Anderson, David E; Löhr, Christiane V

2010-08-01

3 alpaca crias and cadavers of an alpaca cria and a llama cria were evaluated for evidence of esophageal dysfunction. All 5 crias were between 3 and 5 months of age when clinical signs developed, and all had a thin body condition when examined. Clinical signs included coughing, regurgitation, and grossly visible esophageal peristaltic waves. A barium esophagram was used to diagnose esophageal obstruction, megaesophagus, and a vascular ring anomaly (VRA). Fluoroscopy was used to evaluate deglutition, esophageal peristalsis, and the extent of esophageal dilation in 1 alpaca cria. A persistent right aortic arch was identified in 1 alpaca cria, and a left aortic arch with right ductus arteriosus or ligamentum arteriosum and an aberrant right subclavian artery were identified in the 4 remaining crias. Surgical correction of the VRA was attempted in the 3 live alpaca crias. It was complicated by the conformation and location of each VRA and inaccurate anatomic diagnosis of the VRAs before surgery. Treatment was universally unsuccessful because of intraoperative complications and the persistence of clinical signs after surgery. Megaesophagus is typically an idiopathic condition in camelids. However, these findings suggested that camelids with esophageal dysfunction during the neonatal period may have a VRA. The prognosis is grave for camelids with VRA, and accurate anatomic diagnosis of the VRA via the use of advanced imaging techniques (eg, angiography, computed tomography, or magnetic resonance imaging) may improve the success of surgical intervention.

Comparison of fecal egg counting methods in four livestock species.

PubMed

Paras, Kelsey L; George, Melissa M; Vidyashankar, Anand N; Kaplan, Ray M

2018-06-15

Gastrointestinal nematode parasites are important pathogens of all domesticated livestock species. Fecal egg counts (FEC) are routinely used for evaluating anthelmintic efficacy and for making targeted anthelmintic treatment decisions. Numerous FEC techniques exist and vary in precision and accuracy. These performance characteristics are especially important when performing fecal egg count reduction tests (FECRT). The objective of this study was to compare the accuracy and precision of three commonly used FEC methods and determine if differences existed among livestock species. In this study, we evaluated the modified-Wisconsin, 3-chamber (high-sensitivity) McMaster, and Mini-FLOTAC methods in cattle, sheep, horses, and llamas in three phases. In the first phase, we performed an egg-spiking study to assess the egg recovery rate and accuracy of the different FEC methods. In the second phase, we examined clinical samples from four different livestock species and completed multiple replicate FEC using each method. In the last phase, we assessed the cheesecloth straining step as a potential source of egg loss. In the egg-spiking study, the Mini-FLOTAC recovered 70.9% of the eggs, which was significantly higher than either the McMaster (P = 0.002) or Wisconsin (P = 0.002). In the clinical samples from ruminants, Mini-FLOTAC consistently yielded the highest EPG, revealing a significantly higher level of egg recovery (P < 0.0001). For horses and llamas, both McMaster and Mini-FLOTAC yielded significantly higher EPG than Wisconsin (P < 0.0001, P < 0.0001, P < 0.001, and P = 0.024). Mini-FLOTAC was the most accurate method and was the most precise test for both species of ruminants. The Wisconsin method was the most precise for horses and McMaster was more precise for llama samples. We compared the Wisconsin and Mini-FLOTAC methods using a modified technique where both methods were performed using either the Mini-FLOTAC sieve or cheesecloth. The differences in the estimated mean EPG on log scale between the Wisconsin and mini-FLOTAC methods when cheesecloth was used (P < 0.0001) and when cheesecloth was excluded (P < 0.0001) were significant, providing strong evidence that the straining step is an important source of error. The high accuracy and precision demonstrated in this study for the Mini-FLOTAC, suggest that this method can be recommended for routine use in all host species. The benefits of Mini-FLOTAC will be especially relevant when high accuracy is important, such as when performing FECRT. Copyright © 2018 Elsevier B.V. All rights reserved.

Expression, Biochemistry, and Stabilization with Camel Antibodies of Membrane Proteins: Case Study of the Mouse 5-HT3 Receptor.

PubMed

Hassaïne, Ghérici; Deluz, Cédric; Grasso, Luigino; Wyss, Romain; Hovius, Ruud; Stahlberg, Henning; Tomizaki, Takashi; Desmyter, Aline; Moreau, Christophe; Peclinovska, Lucie; Minniberger, Sonja; Mebarki, Lamia; Li, Xiao-Dan; Vogel, Horst; Nury, Hugues

2017-01-01

There is growing interest in the use of mammalian protein expression systems, and in the use of antibody-derived chaperones, for structural studies. Here, we describe protocols ranging from the production of recombinant membrane proteins in stable inducible cell lines to biophysical characterization of purified membrane proteins in complex with llama antibody domains. These protocols were used to solve the structure of the mouse 5-HT3 serotonin receptor but are of broad applicability for crystallization or cryo-electron microscopy projects.

Cerebrospinal Nematodiasis in 20 Camelids.

PubMed

Bertin, F R; Taylor, S D

2016-07-01

Information about the clinical and clinicopathologic aspects of cerebrospinal nematodiasis (CN) in camelids is limited. Clinical and therapeutic variables will be identified as factors predictive of survival. Client-owned camelids suspected of having CN admitted to Purdue University between 1995 and 2015. A retrospective study was performed. A diagnosis of CN was based on cerebrospinal fluid (CSF) eosinophilic pleocytosis or postmortem findings. Eleven alpacas and 9 llamas met the inclusion criteria. Seventy-five percent of the camelids were male (27% castrated and 73% intact). Common clinical abnormalities included proprioceptive deficits (100% of animals), recumbency (55%), tachypnea (55%), and ataxia (40%). Among the 85% of treated animals, 100% received PO fenbendazole, and 88% received a nonsteroidal anti-inflammatory drug. The survival rate to discharge was 45%. Plasma fibrinogen concentration, creatine kinase activity, and serum creatinine concentration were significantly higher in nonsurvivors. Blood eosinophil count, platelet count, and total CO2 were significantly lower in nonsurvivors. Factors associated with survival were species, sex, absence of treatment with corticosteroids, and clinical improvement. There was no association between recumbency at admission and survival. A plasma fibrinogen concentration above >266 mg/dL was an excellent diagnostic test to predict survival in the presence of neurological signs or CSF eosinophilia. Although prognosis for CN in camelids is guarded, presence of recumbency at admission is not predictive of nonsurvival. Male camelids and llamas appear more likely to die from CN. Corticosteroid treatment is contraindicated in animals diagnosed with CN. Copyright © 2016 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

Expression of estrogen receptors alpha and beta in the corpus luteum and uterus from non-pregnant and pregnant llamas.

PubMed

Powell, Susan A; Smith, Bradford B; Timm, Karen I; Menino, Alfred R

2007-08-01

Because estrogen may be involved in maternal recognition of pregnancy and embryonic migration in llamas, expression of estrogen receptor subtypes alpha (ERalpha) and beta (ERbeta) was evaluated in corpus luteum (CL), endometrium, and uterus using relative RT-PCR. Tissues were recovered from sterile-mated (SM) and pregnant (PG) females during Days 7-11 and 7-13 (Day 0 = day of mating), respectively, and follicular phase and juvenile females. Luteal expression of ERalpha and beta was similar (P > 0.10) in SM and PG females and within Days 7-11, however, expression of ERalpha in ovarian tissue from follicular phase females was greater (P < 0.05) than Days 7 and 9 CL. Uterus expressed less ERalpha and beta compared to endometrium (P = 0.07 and P < 0.01, respectively). Expression of ERalpha was greater (P < 0.05) in Day 7 and follicular phase uteri than Days 9 and 11, Day 13 PG and juvenile uteri. Uterine ERbeta expression was greater (P = 0.09) in PG versus SM females and in mated compared to follicular phase females (P < 0.05). Endometrial expression of ERalpha and beta did not differ (P > 0.10) between SM and PG females or by day. The presence of luteal ER during this period may mean a role for estradiol in maternal recognition of pregnancy. Observed increases in uterine ER expression with no changes in endometrium suggest expression increased in myometrium and/or perimetrium. Upregulation of myometrial ERbeta in PG females may be involved in supporting uterine migration of the embryo.

Competitive Selection from Single Domain Antibody Libraries Allows Isolation of High-Affinity Antihapten Antibodies That Are Not Favored in the llama Immune Response

PubMed Central

Rosa, Sofia Tabares-da; Rossotti, Martin; Carleiza, Carmen; Carrión, Federico; Pritsch, Otto; Ahn, Ki Chang; Last, Jerold A.; Hammock, Bruce D; González-Sapienza, Gualberto

2011-01-01

Single-domain antibodies (sdAbs) found in camelids, lack a light chain and their antigen-binding site sits completely in the heavy-chain variable domain (VHH). Their simplicity, thermostability, and ease in expression have made VHHs highly attractive. While this has been successfully exploited for macromolecular antigens, their application to the detection of small molecules is still limited to a very few reports, mostly describing low affinity VHHs. Using triclocarban (TCC) as a model hapten, we found that conventional antibodies, IgG1 fraction, reacted with free TCC with a higher relative affinity (IC50 51.0 ng/mL) than did the sdAbs (IgG2 and IgG3, 497 and 370 ng/mL, respectively). A VHH library was prepared, and by elution of phage with limiting concentrations of TCC and competitive selection of binders, we were able to isolate high-affinity clones, KD 0.98–1.37 nM (SPR) which allowed development of a competitive assay for TCC with an IC50 = 3.5 ng/mL (11 nM). This represents a 100-fold improvement with regard to the performance of the sdAb serum fraction, and it is 100-fold better than the IC50 attained with other anti-hapten VHHs reported thus far. Despite the modest overall anti-hapten sdAbs response in llamas, a small subpopulation of high affinity VHHs are generated that can be isolated by carefully design of the selection process. PMID:21827167

[Pathology of South American Camelids: a retrospective study of necropsies at the Institute of Veterinary Pathology, University of Leipzig, Germany].

PubMed

Theuß, T; Goerigk, D; Rasenberger, S; Starke, A; Schoon, H-A

2014-01-01

The number of South American Camelids (New World Camelids) housed in Germany has increased in the recent years. While these species were formerly kept solely in zoological gardens, ever more private and commercial livestock is being established. Compared to indigenous livestock animals, they bear some distinctive differences, particularly in terms of digestive tract anatomy and physiology. Therefore, it is of considerable interest for veterinarians working with South American Camelids to obtain knowledge about the distinguishing features of these animals and the typical diseases affecting them in Germany. For this purpose, the necropsy reports, including the anamnestic data, and their diagnostic usefulness, from 1995 to 2012 were studied retrospectively. Du- ring this period, a total of 233 New World Camelids were examined (195 alpacas and 38 llamas). Anamnestic data of diagnostic usefulness regarding the cause of disease were only submitted in a limited number of cases, because most of the animals died without specific symptoms. The following were the most frequent pathological findings: enteritis (n = 91), gastritis (n = 76), cachexia (n = 73), pneumonia (n = 30), stomatitis (n = 27), azotaemia (n = 22) and anaemia (n = 9). An endoparasitosis occurred in 107 cases and was considered the predominant cause of enteritis. As with indigenous ruminants, llamas and alpacas primarily suffered from diseases of the digestive and respiratory tracts. Other organ systems were affected to a lesser extent. Even in cases with severe alterations in the affected organs, South American Camelids do not show or show too late diagnostically indicative clinical symptoms. Therefore, a detailed clinical examination of these animals is important.

The Sda/GM2-glycan is a carbohydrate marker of porcine primordial germ cells and of a subpopulation of spermatogonia in cattle, pigs, horses and llama.

PubMed

Klisch, K; Contreras, D A; Sun, X; Brehm, R; Bergmann, M; Alberio, R

2011-11-01

Spermatogonia are a potential source of adult pluripotent stem cells and can be used for testis germ cell transplantation. Markers for the isolation of these cells are of great importance for biomedical applications. Primordial germ cells and prepubertal spermatogonia in many species can be identified by their binding of Dolichos biflorus agglutinin (DBA). This lectin binds to two different types of glycans, which are α-linked N-acetylgalactosamine (GalNac) and β-linked GalNac, if this is part of the Sda or GM2 glycotopes. We used the MAB CT1, which is specific for the trisaccharides motif NeuAcα2-3(GalNAcβ1-4)Galβ1-, which is common to both Sda and GM2 glycotopes, to further define the glycosylation of DBA binding germ cells. In porcine embryos, CT1 bound to migratory germ cells and gonocytes. CT1/DBA double staining showed that the mesonephros was CT1 negative but contained DBA-positive cells. Gonocytes in the female gonad became CT1 negative, while male gonocytes remained CT1 positive. In immunohistological double staining of cattle, pig, horse and llama testis, DBA and CT1 staining was generally colocalised in a subpopulation of spermatogonia. These spermatogonia were mainly single, sometimes paired or formed chains of up to four cells. Our data show that the Sda/GM2 glycotope is present in developing germ cells and spermatogonia in several species. Owing to the narrower specificity of the CT1 antibody, compared with DBA, the former is likely to be a useful tool for labelling and isolation of these cells.

Oxygen binding properties, capillary densities and heart weights in high altitude camelids.

PubMed

Jürgens, K D; Pietschmann, M; Yamaguchi, K; Kleinschmidt, T

1988-01-01

The oxygen binding properties of the blood of the camelid species vicuna, llama, alpaca and dromedary camel were measured and evaluated with respect to interspecific differences. The highest blood oxygen affinity, not only among camelids but of all mammals investigated so far, was found in the vicuna (P50 = 17.6 Torr compared to 20.3-21.6 Torr in the other species). Low hematocrits (23-34%) and small red blood cells (21-30 microns 3) are common features of all camelids, but the lowest values are found in the Lama species. Capillary densities were determined in heart and soleus muscle of vicuna and llama. Again, the vicuna shows exceptional values (3720 cap/mm2 on average in the heart) for a mammal of this body size. Finally, heart weight as percent of body weight is higher in the vicuna (0.7-0.9%) than in the other camelids studied (0.5-0.7%). The possibility that these parameters, measured in New World tylopodes at sea level, are not likely to change considerably with transfer to high altitude, is discussed. In the vicuna, a unique combination of the following features seems to be responsible for an outstanding physical capability at high altitude: saturation of blood with oxygen in the lung is favored by a high blood oxygen affinity, oxygen supply being facilitated by low diffusion distances in the muscle tissue. Loading, as well as unloading, of oxygen is improved by a relatively high oxygen transfer conductance of the red blood cells, which is due to their small size and which compensates the negative effect of a low hematocrit on the oxygen conductance of blood.(ABSTRACT TRUNCATED AT 250 WORDS)

Digesta retention patterns of solute and different-sized particles in camelids compared with ruminants and other foregut fermenters.

PubMed

Dittmann, Marie T; Runge, Ullrich; Ortmann, Sylvia; Lang, Richard A; Moser, Dario; Galeffi, Cordula; Schwarm, Angela; Kreuzer, Michael; Clauss, Marcus

2015-07-01

The mean retention times (MRT) of solute or particles in the gastrointestinal tract and the forestomach (FS) are crucial determinants of digestive physiology in herbivores. Besides ruminants, camelids are the only herbivores that have evolved rumination as an obligatory physiological process consisting of repeated mastication of large food particles, which requires a particle sorting mechanism in the FS. Differences between camelids and ruminants have hardly been investigated so far. In this study we measured MRTs of solute and differently sized particles (2, 10, and 20 mm) and the ratio of large-to-small particle MRT, i.e. the selectivity factors (SF(10/2mm), SF(20/2mm), SF(20/10mm)), in three camelid species: alpacas (Vicugna pacos), llamas (Llama glama), and Bactrian camels (Camelus bactrianus). The camelid data were compared with literature data from ruminants and non-ruminant foregut fermenters (NRFF). Camelids and ruminants both had higher SF(10/2mm)FS than NRFF, suggesting convergence in the function of the FS sorting mechanism in contrast to NRFF, in which such a sorting mechanism is absent. The SF(20/10mm)FS did not differ between ruminants and camelids, indicating that there is a particle size threshold of about 1 cm in both suborders above which particle retention is not increased. Camelids did not differ from ruminants in MRT(2mm)FS, MRTsoluteFS, and the ratio MRT(2mm)FS/MRTsoluteFS, but they were more similar to 'cattle-' than to 'moose-type' ruminants. Camelids had higher SF(10/2mm)FS and higher SF(20/2mm)FS than ruminants, indicating a potentially slower particle sorting in camelids than in ruminants, with larger particles being retained longer in relation to small particles.

Ejaculate fractioning effect on llama sperm head morphometry as assessed by the ISAS(®) CASA system.

PubMed

Soler, C; Sancho, M; García, A; Fuentes, Mc; Núñez, J; Cucho, H

2014-02-01

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 vagina. In five of the animals, a fractioning of the ejaculate was performed by taking samples every 5 min. for a total of 20 min. Air-dried seminal smears were stained with Hemacolor and mounted permanently with Eukitt. Morphometric analysis was carried out with the morphometry module of the ISAS(®) CASA system. Almost 350 cells were analysed per sample, with a total number of 3207 spermatozoa. Mean values were given as follows: length: 5.51 μm; width: 3.38 μm; area: 17.75 μm(2) ; perimeter: 14.8 μm; ellipticity: 0.24; elongation: 0.56; rugosity: 0.87; regularity: 1.07; and shape factor: 1.41. Different animals showed differences in their morphometric values. When we compared the values from different fractions, only two samples showed differences in morphometric parameter values and four samples showed differences in shape parameters. Multivariate analysis allowed the size classification of the cells into three classes and five classes of shapes. The distribution of classes among fractions showed no differences. Despite the individual morphometric differences observed in some fractions, the characteristics of the sperm head morphometry can be considered constant along the ejaculatory period in the llama. © 2013 Blackwell Verlag GmbH.

Evaluation of the acrosomal status in Lama glama sperm incubated with acrosome reaction inducers.

PubMed

Carretero, M I; Fumuso, F G; Neild, D M; Giuliano, S M; Cetica, P; Miragaya, M H

2015-09-01

The objectives of this study were to evaluate the effect of different acrosome reaction (AR) inducers on viability and acrosomal status in llama spermatozoa, by using the FITC-PNA/PI technique and evaluate if there is a positive correlation between the FITC-PNA/PI and the Coomassie blue (CB) staining techniques. After incubating twenty ejaculates in 0.1% collagenase the centrifuged pellets were resuspended in TALP-BSA medium. An aliquot was sonicated to remove the acrosomal content (positive control). The rest of the sample was incubated for 3h at 38 °C with 5% CO2 and 100% humidity. Three aliquots were further incubated 1h with one of the following AR inducers: calcium ionophore, ionomycin or progesterone. One without inducers and the other, incubated with dimethyl sulfoxide (vehicle of the inducing agents). Acrosomes were evaluated at time 0 and after 4h incubation. Calcium ionophore was the most potent agent for inducing the AR (67.2 ± 14.4% live+dead AR sperm) (P < 0.05). These samples showed no motility and viability was very low (0-30%). Both ionomycin and progesterone presented significantly higher (P < 0.05) percentages of total AR sperm than the controls, but had similar percentages of dead reacted sperm to the controls. A positive correlation was observed between the intact acrosome FITC-PNA/PI pattern (live+dead sperm) and the acrosome-present CB pattern (r = 0.64; P = 0.000) in all the evaluated samples. the FITC-PNA/PI technique simultaneously evaluates viability and acrosomal status in llama spermatozoa and calcium ionophore could be used as a control of AR. Copyright © 2015 Elsevier B.V. All rights reserved.

Neosporosis in South America.

PubMed

Moore, D P

2005-01-20

This work gathers reports about Neospora-infections in South America. Neospora-infections have been reported from Argentina, Brazil, Chile, Paraguay, Peru and Uruguay. Evidence of exposure to N. caninum was mentioned in cattle, goats, sheep, dogs, cats, water buffaloes, alpacas, llamas, South American opossums, wolves and other wild canids. No antibodies were found in horses. Interesting epidemiological and pathological data were described. Two isolations were performed from dogs, one from cattle, and recently five from water buffaloes. Since the cattle industry is important in South America and reproductive losses caused by Neospora-infection have been identified, more investigations are needed in order to understand its epidemiology and control the disease.

Hyperglycemia, hypernatremia, and hyperosmolarity in 6 neonatal llamas and alpacas.

PubMed

Cebra, C K

2000-12-01

Neonatal camelids can develop hyperglycemia, hypernatremia, and hyperosmolarity in response to a combination of stress and inadequate water intake. Clinical signs of this syndrome include a fine head tremor, ataxia, and a base-wide stance of the hind limbs, but biochemical analyses are necessary to confirm the diagnosis. Camelids appear to be susceptible to this syndrome because of a poor insulin response to hyperglycemia; hypernatremia results from free water loss associated with glucose diuresis. Water loss associated with glucose diuresis may necessitate a higher rate of fluid administration in camelids with this syndrome than is typically used for treatment of hypernatremia in calves.

Generation of single domain antibody fragments derived from camelids and generation of manifold constructs.

PubMed

Vincke, Cécile; Gutiérrez, Carlos; Wernery, Ulrich; Devoogdt, Nick; Hassanzadeh-Ghassabeh, Gholamreza; Muyldermans, Serge

2012-01-01

Immunizing a camelid (camels and llamas) with soluble, properly folded proteins raises an affinity-matured immune response in the unique camelid heavy-chain only antibodies (HCAbs). The peripheral blood lymphocytes of the immunized animal are used to clone the antigen-binding antibody fragment from the HCAbs in a phage display vector. A representative aliquot of the library of these antigen-binding fragments is used to retrieve single domain antigen-specific binders by successive rounds of panning. These single domain antibody fragments are cloned in tandem to generate manifold constructs (bivalent, biparatopic or bispecific constructs) to increase their functional affinity, to increase specificity, or to connect two independent antigen molecules.

The effect of short duration transportation on serum cortisol response in alpacas (Llama pacos).

PubMed

Anderson, D E; Grubb, T; Silveira, F

1999-03-01

This research project evaluated the changes in serum cortisol in six male and six female alpacas in response to transportation of short duration. All alpacas were subjected to trailer transportation for 30 min. Serum samples were obtained prior to transportation, immediately after transportation, and after a 4-h recovery period. Heart rate was recorded at each time interval and observations of individual behavioural characteristics were recorded. Data were analysed using analysis of variance. Heart rate was not significantly changed by transportation stress. Serum cortisol concentration was significantly higher after transportation, but head returned to baseline concentration after the 4-h recovery period. Behavioural characteristics were not associated with changes in serum cortisol concentration.

Underway Recovery Test 6 (URT-6) - Day 2 Activites

NASA Image and Video Library

2018-01-18

As part of Underway Recovery Test 6, the Orion test article is pulled in by a winch line at the rear of the USS Anchorage’s well deck that brings the capsule into the ship, along with four manned LLAMAs (Line Load Attenuation Mechanism Assembly) that control the capsule’s side-to-side movement and a tending line attached to a rigid hull inflatable boat for controlling Orion’s movement behind the ship. The testing with Kennedy Space Center's NASA Recovery Team and the U.S. Navy will provide important data that is being used to improve recovery procedures and hardware ahead of Orion's next flight, Exploration Mission-1, when it splashes down in the Pacific Ocean.

Tilting the balance between canonical and noncanonical conformations for the H1 hypervariable loop of a llama VHH through point mutations.

PubMed

Mahajan, Sai Pooja; Velez-Vega, Camilo; Escobedo, Fernando A

2013-01-10

Nanobodies are single-domain antibodies found in camelids. These are the smallest naturally occurring binding domains and derive functionality via three hypervariable loops (H1-H3) that form the binding surface. They are excellent candidates for antibody engineering because of their favorable characteristics like small size, high solubility, and stability. To rationally engineer antibodies with affinity for a specific target, the hypervariable loops can be tailored to obtain the desired binding surface. As a first step toward such a goal, we consider the design of loops with a desired conformation. In this study, we focus on the H1 loop of the anti-hCG llama nanobody that exhibits a noncanonical conformation. We aim to "tilt" the stability of the H1 loop structure from a noncanonical conformation to a (humanized) type 1 canonical conformation by studying the effect of selected mutations to the amino acid sequence of the H1, H2, and proximal residues. We use all-atomistic, explicit-solvent, biased molecular dynamic simulations to simulate the wild-type and mutant loops in a prefolded framework. We thus find mutants with increasing propensity to form a stable type 1 canonical conformation of the H1 loop. Free energy landscapes reveal the existence of conformational isomers of the canonical conformation that may play a role in binding different antigenic surfaces. We also elucidate the approximate mechanism and kinetics of transitions between such conformational isomers by using a Markovian model. We find that a particular three-point mutant has the strongest thermodynamic propensity to form the H1 type 1 canonical structure but also to exhibit transitions between conformational isomers, while a different, more rigid three-point mutant has the strongest propensity to be kinetically trapped in such a canonical structure.

Ebolavirus Nucleoprotein C-Termini Potently Attract Single Domain Antibodies Enabling Monoclonal Affinity Reagent Sandwich Assay (MARSA) Formulation

PubMed Central

Sherwood, Laura J.; Hayhurst, Andrew

2013-01-01

Background Antigen detection assays can play an important part in environmental surveillance and diagnostics for emerging threats. We are interested in accelerating assay formulation; targeting the agents themselves to bypass requirements for a priori genome information or surrogates. Previously, using in vitro affinity reagent selection on Marburg virus we rapidly established monoclonal affinity reagent sandwich assay (MARSA) where one recombinant antibody clone was both captor and tracer for polyvalent nucleoprotein (NP). Hypothesizing that the closely related Ebolavirus genus may share the same Achilles' heel, we redirected the scheme to see whether similar assays could be delivered and began to explore their mechanism. Methods and Findings In parallel we selected panels of llama single domain antibodies (sdAb) from a semi-synthetic library against Zaire, Sudan, Ivory Coast, and Reston Ebola viruses. Each could perform as both captor and tracer in the same antigen sandwich capture assay thereby forming MARSAs. All sdAb were specific for NP and those tested required the C-terminal domain for recognition. Several clones were cross-reactive, indicating epitope conservation across the Ebolavirus genus. Analysis of two immune shark sdAb revealed they also targeted the C-terminal domain, and could be similarly employed, yet were less sensitive than a comparable llama sdAb despite stemming from immune selections. Conclusions The C-terminal domain of Ebolavirus NP is a strong attractant for antibodies and enables sensitive sandwich immunoassays to be rapidly generated using a single antibody clone. The polyvalent nature of nucleocapsid borne NP and display of the C-terminal region likely serves as a bountiful affinity sink during selections, and a highly avid target for subsequent immunoassay capture. Combined with the high degree of amino acid conservation through 37 years and across wide geographies, this domain makes an ideal handle for monoclonal affinity reagent driven antigen sandwich assays for the Ebolavirus genus. PMID:23577211

16 kDa heat shock protein from heat-inactivated Mycobacterium tuberculosis is a homodimer - suitability for diagnostic applications with specific llama VHH monoclonals.

PubMed

Srivastava, Saurabh K; Ruigrok, Vincent J B; Thompson, Natalie J; Trilling, Anke K; Heck, Albert J R; van Rijn, Cees; Beekwilder, Jules; Jongsma, Maarten A

2013-01-01

The 16 kDa heat shock protein (HSP) is an immuno-dominant antigen, used in diagnosis of infectious Mycobacterium tuberculosis (M.tb.) causing tuberculosis (TB). Its use in serum-based diagnostics is limited, but for the direct identification of M.tb. bacteria in sputum or cultures it may represent a useful tool. Recently, a broad set of twelve 16 kDa specific heavy chain llama antibodies (VHH) has been isolated, and their utility for diagnostic applications was explored. To identify the epitopes recognized by the nine (randomly selected from a set of twelve 16 kDa specific VHH antibodies) distinct VHH antibodies, 14 overlapping linear epitopes (each 20 amino acid long) were characterized using direct and sandwich ELISA techniques. Seven out of 14 epitopes were recognized by 8 out of 9 VHH antibodies. The two highest affinity binders B-F10 and A-23 were found to bind distinct epitopes. Sandwich ELISA and SPR experiments showed that only B-F10 was suitable as secondary antibody with both B-F10 and A-23 as anchoring antibodies. To explain this behavior, the epitopes were matched to the putative 3D structure model. Electrospray ionization time-of-flight mass spectrometry and size exclusion chromatography were used to determine the higher order conformation. A homodimer model best explained the differential immunological reactivity of A-23 and B-F10 against heat-treated M.tb. lysates. The concentrations of secreted antigens of M.tb. in sputum are too low for immunological detection and existing kits are only used for identifying M.tb. in cultures. Here we describe how specific combinations of VHH domains could be used to detect the intracellular HSP antigen. Linked to methods of pre-concentrating M.tb. cells prior to lysis, HSP detection may enable the development of protein-based diagnostics of sputum samples and earlier diagnosis of diseases.

Unveiling a Drift Resistant Cryptotope within Marburgvirus Nucleoprotein Recognized by Llama Single-Domain Antibodies

PubMed Central

Garza, John Anthony; Taylor, Alexander Bryan; Sherwood, Laura Jo; Hart, Peter John; Hayhurst, Andrew

2017-01-01

Marburg virus (MARV) is a highly lethal hemorrhagic fever virus that is increasingly re-emerging in Africa, has been imported to both Europe and the US, and is also a Tier 1 bioterror threat. As a negative sense RNA virus, MARV has error prone replication which can yield progeny capable of evading countermeasures. To evaluate this vulnerability, we sought to determine the epitopes of 4 llama single-domain antibodies (sdAbs or VHH) specific for nucleoprotein (NP), each capable of forming MARV monoclonal affinity reagent sandwich assays. Here, we show that all sdAb bound the C-terminal region of NP, which was produced recombinantly to derive X-ray crystal structures of the three best performing antibody-antigen complexes. The common epitope is a trio of alpha helices that form a novel asymmetric basin-like depression that accommodates each sdAb paratope via substantial complementarity-determining region (CDR) restructuring. Shared core contacts were complemented by unique accessory contacts on the sides and overlooks of the basin yielding very different approach routes for each sdAb to bind the antigen. The C-terminal region of MARV NP was unable to be crystallized alone and required engagement with sdAb to form crystals suggesting the antibodies acted as crystallization chaperones. While gross structural homology is apparent between the two most conserved helices of MARV and Ebolavirus, the positions and morphologies of the resulting basins were markedly different. Naturally occurring amino acid variations occurring in bat and human Marburgvirus strains all mapped to surfaces distant from the predicted sdAb contacts suggesting a vital role for the NP interface in virus replication. As an essential internal structural component potentially interfacing with a partner protein it is likely the C-terminal epitope remains hidden or “cryptic” until virion disruption occurs. Conservation of this epitope over 50 years of Marburgvirus evolution should make these sdAb useful foundations for diagnostics and therapeutics resistant to drift. PMID:29038656

Validation of the FAMACHA© system in South American camelids.

PubMed

Storey, Bobby E; Williamson, Lisa H; Howell, Sue B; Terrill, Thomas H; Berghaus, Roy; Vidyashankar, Anand N; Kaplan, Ray M

2017-08-30

Haemonchus contortus resistant to multiple anthelmintics threaten the viability of the small ruminant industry in areas where this parasite is prevalent. In response to this situation, the FAMACHA© system was developed and validated for use with small ruminants as a way to detect clinical anemia associated with haemonchosis. Given that H. contortus and multiple anthelmintic resistance is a similar problem in camelids, the FAMACHA© system might also provide the same benefits. To address this need, a validation study of the FAMACHA© system was conducted on 21 alpaca and llama farms over a 2-year period. H. contortus was the predominant nematode parasite on 17 of the 21 farms (10 alpaca and 7 llama farms) enrolled in the study, based on fecal culture results. The FAMACHA© card was used to score the color of the lower palpebral (lower eye lid) conjunctiva on a 1-5 scale. Packed cell volume (PCV) values were measured and compared to FAMACHA© scores using FAMACHA© score cutoffs of ≥3 or ≥4 and with anemia defined as a PCV ≤15%, ≤17%, or≤20%. PCV was significantly associated with FAMACHA© score, fecal egg count (FEC), and body condition score (BCS), regardless of the FAMACHA© cutoff score or the PCV% chosen to define clinical anemia (p<0.01 in all cases). The use of FAMACHA© scores ≥3 and PCV ≥ 15% indicating anemia provided the best sensitivity (96.4% vs 92.9% for FAMACHA© ≥4), whereas FAMACHA scores ≥ 4 and PCV ≤20% provided the best specificity (94.2% vs 69.1% for FAMACHA© ≥3). The data from this study support the FAMACHA© system as a useful tool for detecting clinical anemia in camelids suffering from haemonchosis. Parameters for making treatment decisions based on FAMACHA© score in camelids should mirror those established for small ruminants. Copyright © 2017 Elsevier B.V. All rights reserved.

The Glycosylphosphatidylinositol-Anchored Variable Region of Llama Heavy Chain-Only Antibody JM4 Efficiently Blocks both Cell-Free and T Cell-T Cell Transmission of Human Immunodeficiency Virus Type 1

PubMed Central

Liu, Lihong; Wang, Weiming; Matz, Julie; Ye, Chaobaihui; Bracq, Lucie; Delon, Jerome; Kimata, Jason T.; Chen, Zhiwei

2016-01-01

ABSTRACT The variable regions (VHHs) of two heavy chain-only antibodies, JM2 and JM4, from llamas that have been immunized with a trimeric gp140 bound to a CD4 mimic have been recently isolated (here referred to as VHH JM2 and VHH JM4, respectively). JM2 binds the CD4-binding site of gp120 and neutralizes HIV-1 strains from subtypes B, C, and G. JM4 binds gp120 and neutralizes HIV-1 strains from subtypes A, B, C, A/E, and G in a CD4-dependent manner. In the present study, we constructed glycosylphosphatidylinositol (GPI)-anchored VHH JM2 and JM4 along with an E4 control and transduced them into human CD4+ cell lines and primary CD4 T cells. We report that by genetically linking the VHHs with a GPI attachment signal, VHHs are targeted to the lipid rafts of the plasma membranes. Expression of GPI-VHH JM4, but not GPI-VHH E4 and JM2, on the surface of transduced TZM.bl cells potently neutralizes multiple subtypes of HIV-1 isolates, including tier 2 or 3 strains, transmitted founders, quasispecies, and soluble single domain antibody (sdAb) JM4-resistant viruses. Moreover, transduction of CEMss-CCR5 cells with GPI-VHH JM4, but not with GPI-VHH E4, confers resistance to both cell-free and T cell-T cell transmission of HIV-1 and HIV-1 envelope-mediated fusion. Finally, GPI-VHH JM4-transduced human primary CD4 T cells efficiently resist both cell-free and T cell-T cell transmission of HIV-1. Thus, we conclude that VHH JM4, when targeted to the lipid rafts of the plasma membrane, efficiently neutralizes HIV-1 infection via both cell-free and T cell-T cell transmission. Our findings should have important implications for GPI-anchored antibody-based therapy against HIV-1. IMPORTANCE Lipid rafts are specialized dynamic microdomains of the plasma membrane and have been shown to be gateways for HIV-1 budding as well as entry into T cells and macrophages. In nature, many glycosylphosphatidylinositol (GPI)-anchored proteins localize in the lipid rafts. In the present study, we developed GPI-anchored variable regions (VHHs) of two heavy chain-only antibodies, JM2 and JM4, from immunized llamas. We show that by genetically linking the VHHs with a GPI attachment signal, VHHs are targeted to the lipid rafts of the plasma membranes. GPI-VHH JM4, but not GPI-VHH JM2, in transduced CD4+ cell lines and human primary CD4 T cells not only efficiently blocks diverse HIV-1 strains, including tier 2 or 3 strains, transmitted founders, quasispecies, and soluble sdAb JM4-resistant strains, but also efficiently interferes T cell-T cell transmissions of HIV-1 and HIV-1 envelope-mediated fusion. Our findings should have important implications in GPI-anchored antibody-based therapy against HIV-1. PMID:27654286

[Suckling behavior of llamas (Lama glama)].

PubMed

Gauly, M; Pouillion, C; Erhardt, G

2003-10-01

In the present study the suckling behaviour of six Ilama crias was analysed. A total of 3398 suckling bouts were observed and evaluated. The crias mostly suckled in a reverse parallel position with no side preference. The cria initiated suckling in most suckling bouts whereas the dams usually ended them. The total number of suckling bouts could be divided into two types: suckling bouts of short duration and suckling bouts of long duration. There were no significant differences in duration per suckling bout among the different ages of crias. But the animals suckled less frequently and spent less total time suckling as they grow older. The age related differences and evidence for a dual function (nutrition, social contact) of suckling behaviour are discussed.

[Competence in biocentric ethics.

PubMed

Stepke, Fernando Lolas

2012-07-01

ResumenEste artículo explora el origen y la conceptualización de la bioética en tanto ética biocéntrica, definida por el pionero Fritz Jahr como un imperativo hipotético que llama a respetar la vida en todas sus formas. Tras describirla como proceso social, procedimiento técnico y producto académico, se enumeran y comentan algunas competencias que debieran presidir el empleo del discurso bioético en cualquier punto del complejo social "investigación médica": miembros de comités de ética de investigación, investigadores, empresarios, administradores. Cada uno de estos grupos, en su "cultura epistémica" propia, define y explicita los principios básicos en tanto expresión de valores y leyes generales.

Underway Recovery Test 6 (URT-6) - Day 2 Activites

NASA Image and Video Library

2018-01-18

Off the rear of the USS Anchorage, the Orion test article is pulled in by a winch line at the rear of the USS Anchorage’s well deck that brings the capsule into the ship, along with four manned LLAMAs (Line Load Attenuation Mechanism Assembly) that control the capsule’s side-to-side movement and a tending line attached to a rigid hull inflatable boat for controlling Orion’s movement behind the ship. The Underway Recovery Test 6 (URT-6) is spearheaded by Kennedy Space Center's NASA Recovery Team. In partnership with the U.S. Navy, the testing will provide important data that is being used to improve recovery procedures and hardware ahead of Orion's next flight, Exploration Mission-1, when it splashes down in the Pacific Ocean.

Competencias en ética biocéntrica

PubMed Central

Stepke, Fernando Lolas

2013-01-01

Resumen Este artículo explora el origen y la conceptualización de la bioética en tanto ética biocéntrica, definida por el pionero Fritz Jahr como un imperativo hipotético que llama a respetar la vida en todas sus formas. Tras describirla como proceso social, procedimiento técnico y producto académico, se enumeran y comentan algunas competencias que debieran presidir el empleo del discurso bioético en cualquier punto del complejo social “investigación médica”: miembros de comités de ética de investigación, investigadores, empresarios, administradores. Cada uno de estos grupos, en su “cultura epistémica” propia, define y explicita los principios básicos en tanto expresión de valores y leyes generales. PMID:23459767

Long-bone fractures in llamas and alpacas: 28 cases (1998–2008)

PubMed Central

Knafo, S. Emmanuelle; Getman, Liberty M.; Richardson, Dean W.; Fecteau, Marie-Eve

2012-01-01

Treatment and outcome of camelids with long-bone fractures are described. Medical records (1998–2008) of camelids (n = 28) with long-bone fractures were reviewed for signalment, time to presentation, fracture type, method of repair, duration of hospitalization, and post-operative complications. Follow-up information was obtained via telephone interviews with owners. Mean age and weight at presentation were 3.4 years and 56.3 kg, respectively. Twenty-six fractures were treated with internal fixation (n = 11), external fixation (n = 10), combination of internal and external fixation (n = 3), amputation (n = 1), and external fixation followed by amputation (n = 1). Long-term follow-up information was obtained for 19 of the 26 animals. The post-operative complication rate was 23% and owner satisfaction was high. Animals with open fractures were more likely to experience complications. Internal fixation was associated with superior alignment and outcome. Internal fixation techniques should be recommended for camelids. PMID:23277645

Ovarian dysgenesis in an alpaca with a minute chromosome 36.

PubMed

Fellows, Elizabeth; Kutzler, Michelle; Avila, Felipe; Das, Pranab J; Raudsepp, Terje

2014-01-01

A 4-year-old female alpaca (Lama pacos [LPA]) was presented to the Oregon State Veterinary Teaching Hospital for failure to display receptive behavior to males. Although no abnormalities were found on physical examination, transrectal ultrasonographic examination of the reproductive tract revealed uterine hypoplasia and ovarian dysgenesis. Cytogenetic analysis demonstrated a normal female 74,XX karyotype with 1 exceptionally small (minute) homologue of autosome LPA36. Chromosome analysis by Giemsa staining and DAPI- and C-banding revealed that the minute LPA36 was submetacentric, AT-rich, and largely heterochromatic. Because of the small size and lack of molecular markers, it was not possible to identify the origin of the minute. There is a need to improve molecular cytogenetic tools to further study the phenomenon of this minute chromosome and its relation to female reproduction in alpacas and llamas. © The American Genetic Association. 2012. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Reproductive glycogenetics--a critical factor in pregnancy success and species hybridisation.

PubMed

Jones, C J P; Aplin, J D

2009-03-01

Hybridisation occurs rarely in nature and experiments using interspecific transfer of embryos generally result in implantation failure. Here we show that appropriate glycosylation of the apposing surfaces of endometrium and trophoblast probably is an important factor and may play a critical role in pregnancy success. Examination of closely related species shows that each has its own specific pattern of glycosylation, or glycotype, at the fetomaternal interface and that interacting surfaces appear to show complementarity, suggesting the existence of a glycocode. Studies on a camel/llama hybrid show that for successful implantation to occur, a hybrid must have a placental glycosylation pattern similar to that of the host species, suggesting that the glycocode and appropriate glycosylation may be critical factors in the establishment and maintenance of pregnancy. This new field of reproductive glycogenetics is not only relevant to the development of new species but may also have important implications in the area of human fertility.

Long-bone fractures in llamas and alpacas: 28 cases (1998-2008).

PubMed

Knafo, S Emmanuelle; Getman, Liberty M; Richardson, Dean W; Fecteau, Marie-Eve

2012-07-01

Treatment and outcome of camelids with long-bone fractures are described. Medical records (1998-2008) of camelids (n = 28) with long-bone fractures were reviewed for signalment, time to presentation, fracture type, method of repair, duration of hospitalization, and post-operative complications. Follow-up information was obtained via telephone interviews with owners. Mean age and weight at presentation were 3.4 years and 56.3 kg, respectively. Twenty-six fractures were treated with internal fixation (n = 11), external fixation (n = 10), combination of internal and external fixation (n = 3), amputation (n = 1), and external fixation followed by amputation (n = 1). Long-term follow-up information was obtained for 19 of the 26 animals. The post-operative complication rate was 23% and owner satisfaction was high. Animals with open fractures were more likely to experience complications. Internal fixation was associated with superior alignment and outcome. Internal fixation techniques should be recommended for camelids.

Carpal valgus in llamas and alpacas: Retrospective evaluation of patient characteristics, radiographic features and outcomes following surgical treatment

PubMed Central

Hunter, Barbara; Duesterdieck-Zellmer, Katja F.; Huber, Michael J.; Parker, Jill E.; Semevolos, Stacy A.

2014-01-01

This study evaluated outcomes of surgical treatment for carpal valgus in New World camelids and correlated successful outcome (absence of carpal valgus determined by a veterinarian) with patient characteristics and radiographic features. Univariable and multivariable analyses of retrospective case data in 19 camelids (33 limbs) treated for carpal valgus between 1987 and 2010 revealed that procedures incorporating a distal radial transphyseal bridge were more likely (P = 0.03) to result in success after a single surgical procedure. A greater degree of angulation (> 19°, P = 0.02) and younger age at surgery (< 4 months, P = 0.03) were associated with unsuccessful outcome. Overall, 74% of limbs straightened, 15% overcorrected, and 11% had persistent valgus following surgical intervention. To straighten, 22% of limbs required multiple procedures, not including implant removal. According to owners, valgus returned following implant removal in 4 limbs that had straightened after surgery. PMID:25477542

Oriented conjugation of single-domain antibodies and quantum dots.

PubMed

Brazhnik, Kristina; Nabiev, Igor; Sukhanova, Alyona

2014-01-01

Nanoparticle-based biodetection routinely employs monoclonal antibodies (mAbs) for targeting. However, the large size of mAbs limits the number of ligands per nanoparticle and severely restricts the bioavailability and distribution of these probes in a sample. Furthermore, conventional conjugation techniques provide nanoprobes with irregular orientation of mAbs on the nanoparticle surface and often provoke mAb unfolding. Here, we describe a protocol for engineering a new generation of ultrasmall diagnostic nanoprobes through oriented conjugation of semiconductor quantum dots (QDs) with 13 kDa single-domain antibodies (sdAbs) derived from llama immunoglobulin G (IgG). The sdAbs are conjugated with QDs in a highly oriented manner via an additional cysteine residue specifically integrated into the sdAb C-terminus. The resultant nanoprobes are <12 nm in diameter, ten times smaller in volume compared to the known alternatives. They have been proved highly efficient in flow cytometry and immunuhistochemical diagnostics. This approach can be easily extended to other semiconductor and plasmonic nanoparticles.

Human llamas: adaptation to altitude in subjects with high hemoglobin oxygen affinity.

PubMed Central

Hebbel, R P; Eaton, J W; Kronenberg, R S; Zanjani, E D; Moore, L G; Berger, E M

1978-01-01

To assess the adaptive value of the right-shift of the oxyhemoglobin dissociation curve (decreased affinity for oxygen) observed in humans upon altitude exposure, the short-term physiologic responses to altitude-induced hypoxia were evaluated in two subjects with a high oxygen affinity hemoglobin (Hb Andrew-Minneapolis) and in two of their normal siblings. In striking contrast to normal subjects, at moderately high altitude (3,100 m) the high affinity subjects manifested: (a) lesser increments in resting heart rate; (b) minimal increases in plasma and urinary erythropoietin; (c) no decrement in maximal oxygen consumption; and (d) no thrombocytopenia. There was no difference between subject pairs in 2,3-diphosphoglycerate response to altitude exposure. These results tend to contradict the belief that a decrease in hemoglobin oxygen affinity is of adaptive value to humans at moderate altitudes. Rather, they support the hypothesis that, despite disadvantages at low altitude, a left-shifted oxyhemoglobin dissociation curve may confer a degree of preadaptation to altitude. PMID:29054

Mapping the Extent and Stability of Montane Wetlands ("Bofedales") in the Andes of Bolivia and Peru using Landsat Imagery, 1980s to Present.

NASA Astrophysics Data System (ADS)

Slayback, D. A.; Hubbard, A. B.; Yager, K.

2017-12-01

Montane wetlands (locally termed "bofedales") in the Andean highlands of Peru and Bolivia provide an important forage resource for camelids (llama and alpaca) in extensive pastoral agriculture systems. For many communities, these systems may provide the only available green forage during the long winter dry season. As a part of a larger project, we are examining the impacts of both current climate change and of socioeconomic drivers on the health and status of these systems. A key part of this analysis requires mapping the current bofedal extent, and observed changes over the recent past, to link to hypothesized drivers. We are therefore mapping bofedal extent using Landsat imagery, from the mid 1980s to present, using a variety of methods (supervised and unsupervised classification, and machine learning). We will present preliminary results from this analysis, showing total regional extent, and observed stability of the bofedales (or lack thereof) over time.

High altitude artisanal small-scale gold mines are hot spots for Mercury in soils and plants.

PubMed

Terán-Mita, Tania A; Faz, Angel; Salvador, Flor; Arocena, Joselito M; Acosta, Jose A

2013-02-01

Mercury releases from artisanal and small-scale gold mines (ASGM) condense and settle on plants, soils and water bodies. We collected soil and plant samples to add knowledge to the likely transfer of Hg from soils into plants and eventually predict Hg accumulation in livestock around ASGM in Bolivia. Mean contents of Hg in soils range from 0.5 to 48.6 mg Hg kg(-1) soil (5× to 60× more compared to control sites) and exceeded the soil Hg threshold levels in some European countries. The Hg contents ranged from 0.6 to 18 and 0.2 to 28.3 mg Hg kg(-1) leaf and root, respectively. The high Hg in Poaceae and Rosaceae may elevate Hg accumulation into the food chain because llama and alpaca solely thrive on these plants for food. Erosion of soils around ASGM in Bolivia contributes to the Hg contamination in lower reaches of the Amazon basin. Copyright © 2012 Elsevier Ltd. All rights reserved.

Cranial arterial patterns of the alpaca (Camelidae: Vicugna pacos).

PubMed

O'Brien, Haley D

2017-03-01

Artiodactyl cranial arterial patterns deviate significantly from the standard mammalian pattern, most notably in the possession of a structure called the carotid rete (CR)-a subdural arterial meshwork that is housed within the cavernous venous sinus, replacing the internal carotid artery (ICA). This relationship between the CR and the cavernous sinus facilitates a suite of unique physiologies, including selective brain cooling. The CR has been studied in a number of artiodactyls; however, to my knowledge, only a single study to date documents a subset of the cranial arteries of New World camelids (llamas, alpacas, vicugñas and guanacoes). This study is the first complete description of the cranial arteries of a New World camelid species, the alpaca ( Vicugna pacos ), and the first description of near-parturition cranial arterial morphology within New World camelids. This study finds that the carotid arterial system is conserved between developmental stages in the alpaca, and differs significantly from the pattern emphasized in other long-necked ruminant artiodactyls in that a patent, homologous ICA persists through the animal's life.

Cranial arterial patterns of the alpaca (Camelidae: Vicugna pacos)

PubMed Central

2017-01-01

Artiodactyl cranial arterial patterns deviate significantly from the standard mammalian pattern, most notably in the possession of a structure called the carotid rete (CR)—a subdural arterial meshwork that is housed within the cavernous venous sinus, replacing the internal carotid artery (ICA). This relationship between the CR and the cavernous sinus facilitates a suite of unique physiologies, including selective brain cooling. The CR has been studied in a number of artiodactyls; however, to my knowledge, only a single study to date documents a subset of the cranial arteries of New World camelids (llamas, alpacas, vicugñas and guanacoes). This study is the first complete description of the cranial arteries of a New World camelid species, the alpaca (Vicugna pacos), and the first description of near-parturition cranial arterial morphology within New World camelids. This study finds that the carotid arterial system is conserved between developmental stages in the alpaca, and differs significantly from the pattern emphasized in other long-necked ruminant artiodactyls in that a patent, homologous ICA persists through the animal's life. PMID:28405385

Alpaca (Lama pacos) as a convenient source of recombinant camelid heavy chain antibodies (VHHs)

PubMed Central

Maass, David R.; Sepulveda, Jorge; Pernthaner, Anton; Shoemaker, Charles B.

2007-01-01

Recombinant single domain antibody fragments (VHHs) that derive from the unusual camelid heavy chain only IgG class (HCAbs) have many favourable properties compared with single-chain antibodies prepared from conventional IgG. As a result, VHHs have become widely used as binding reagents and are beginning to show potential as therapeutic agents. To date, the source of VHH genetic material has been camels and llamas despite their large size and limited availability. Here we demonstrate that the smaller, more tractable and widely available alpaca is an excellent source of VHH coding DNA. Alpaca sera IgG consists of about 50% HCAbs, mostly of the short-hinge variety. Sequencing of DNA encoding more than 50 random VHH and hinge domains permitted the design of PCR primers that will amplify virtually all alpaca VHH coding DNAs for phage display library construction. Alpacas were immunized with ovine tumour necrosis factor α (TNFα) and a VHH phage display library was prepared from a lymph node that drains the sites of immunizations and successfully employed in the isolation of VHHs that bind and neutralize ovine TNFα. PMID:17568607

Oxygen transport of hemoglobin in high-altitude animals (Camelidae).

PubMed

Reynafarje, C; Faura, J; Villavicencio, D; Curaca, A; Reynafarje, B; Oyola, L; Contreras, L; Vallenas, E; Faura, A

1975-05-01

To clarify the mechanisms by which high-altitude Camelidae can adapt to hypoxia, the study of some blood characteristics were carried out in apacas and llamas. The results show that there is a peculiar dissociation curve of hemoglobin in alpacas which permits great affinity of hemoglobin for oxygen at lung level and the release of oxygen at the tissue level with a facility similar to that in man. Fetal hemoglobin was found high in adult alpacas (55 percent). Electrophoretic studies of hemoglobin showed that this pigment has two components, both of which have a very low mobility. Lactic dehydrogenase was found six times higher than in humans. RBC glucose-6-phosphate dehydrogenase was two times higher than in man living at the same altitude. Myoglobin was found to be higher than in man living at altitude. Alpacas have erythrocytes in which the amount of 2,3-DPG is approximately the same as in man. RBC are more resistent to hypotonic solutions than humans. The amount of lactic dehydrogenase, myoglobin, and glucose-6-phosphate dehydrogenase dimishes when alpacas are bought down to sea level.

Single-domain antibody bioconjugated near-IR quantum dots for targeted cellular imaging of pancreatic cancer.

PubMed

Zaman, Md Badruz; Baral, Toya Nath; Jakubek, Zygmunt J; Zhang, Jianbing; Wu, Xiaohua; Lai, Edward; Whitfield, Dennis; Yu, Kui

2011-05-01

Successful targeted imaging of BxPC3 human pancreatic cancer cells is feasible with near-IR CdTeSe/CdS quantum dots (QDs) functionalized with single-domain antibody (sdAb) 2A3. For specific targeting, sdAbs are superior to conventional antibodies, especially in terms of stability, aggregation, and production cost. The bright CdTeSe/CdS QDs were synthesized to emit in the diagnostic window of 650-900 nm with a narrow emission band. 2A3 was derived from llama and is small in size of 13 kDa, but with fully-functional recognition to the target carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6), a possible biomarker as a therapeutic target of pancreatic cancer. For compelling imaging, optical may be the most sensible among the various imaging modalities, regarding the sensitivity and cost. This first report on sdAb-conjugated near-IR QDs with high signal to background sensitivity for targeted cellular imaging brings insights into the development of optical molecular imaging for early stage cancer diagnosis.

NaLi-H1: A universal synthetic library of humanized nanobodies providing highly functional antibodies and intrabodies

PubMed Central

Moutel, Sandrine; Bery, Nicolas; Bernard, Virginie; Keller, Laura; Lemesre, Emilie; de Marco, Ario; Ligat, Laetitia; Rain, Jean-Christophe; Favre, Gilles; Olichon, Aurélien; Perez, Franck

2016-01-01

In vitro selection of antibodies allows to obtain highly functional binders, rapidly and at lower cost. Here, we describe the first fully synthetic phage display library of humanized llama single domain antibody (NaLi-H1: Nanobody Library Humanized 1). Based on a humanized synthetic single domain antibody (hs2dAb) scaffold optimized for intracellular stability, the highly diverse library provides high affinity binders without animal immunization. NaLi-H1 was screened following several selection schemes against various targets (Fluorescent proteins, actin, tubulin, p53, HP1). Conformation antibodies against active RHO GTPase were also obtained. Selected hs2dAb were used in various immunoassays and were often found to be functional intrabodies, enabling tracking or inhibition of endogenous targets. Functionalization of intrabodies allowed specific protein knockdown in living cells. Finally, direct selection against the surface of tumor cells produced hs2dAb directed against tumor-specific antigens further highlighting the potential use of this library for therapeutic applications. DOI: http://dx.doi.org/10.7554/eLife.16228.001 PMID:27434673

Pancreatic necrosis in New World camelids: 11 cases (1990-1998).

PubMed

Pearson, E G; Snyder, S P

2000-07-15

To determine clinical, clinicopathologic, and postmortem abnormalities in New World camelids with pancreatic necrosis. Retrospective study. 10 llamas and 1 alpaca. Medical records of animals in which a diagnosis of pancreatic necrosis had been made on the basis of histologic examination of necropsy specimens or on the basis of clinical signs and results of clinicopathologic testing were reviewed. The initial owner complaint varied, and various other conditions were diagnosed. Clinical and clinicopathologic abnormalities were vague. Amylase activity was higher in abdominal fluid than in serum in 5 of 7 animals, and lipase activity was higher in abdominal fluid than in serum in all 7. Four animals survived, and 7 died or were euthanatized. Only 1 of the animals that died had marked inflammation of the pancreatic parenchyma. All 7 had necrosis and saponification of fat in and surrounding the pancreas. Results suggest that pancreatic necrosis may develop in New World camelids, but clinical signs are vague, and the condition may easily be confused with other diseases. The only laboratory test that appeared to be helpful in the antemortem diagnosis of pancreatic necrosis was comparison of amylase and lipase activities in abdominal fluid and serum.

Gastrointestinal and blood parasite determination in the guanaco (Lama guanicoe) under semi-captivity conditions.

PubMed

Correa, Loreto; Zapata, Beatriz; Soto-Gamboa, Mauricio

2012-01-01

The breeding of wild animals for commercial purposes is becoming more frequent nowadays. This situation has led to an increase in contact rates between wild and domestic animals, with subsequent reciprocal transmission of parasites. In this study, we characterized the gastrointestinal and blood parasites of a group of 15 semi-captive guanacos (Lama guanicoe). We characterized gastrointestinal parasites by analyzing fecal samples through the sedimentation-flotation technique and hemoparasites by using blood smears stained with Giemsa. We found several gastrointestinal parasites including Nematoda and protozoans. The most frequently found parasites were Nematodirus sp. and Eimeria sp. In contrast with previous studies, neither Cestoda nor Fasciola were found. The only hemoparasite detected was Mycoplasma haemolamae, a parasite already described in llamas and alpacas. We conclude that the most frequent gastrointestinal parasites of semi-captive guanacos were nematodes and protozoans. Also, the hemoparasite M. haemolamae seems to be prevalent among captive populations of South American camelids. Finally, captive guanacos share several parasites with the traditional livestock. Therefore, keeping captive or semi-captive guanacos without an adequate sanitary protocol might have adverse consequences to adjacent traditional cattle farming and/or for wild animals.

Bioaugmentation of the anaerobic digestion of food waste by dungs of herbivore, carnivore, and omnivore zoo animals.

PubMed

Ariunbaatar, Javkhlan; Ozcan, Onur; Bair, Robert; Esposito, Giovanni; Ball, Ray; Lens, Piet N L; Yeh, Daniel H

2018-02-01

The potential improvement of biomethanation of food waste (FW) by adding dung of herbivore (giraffe, llama, koala), carnivore (tiger), and omnivore (sloth bear) animals to anaerobic sludge (AnS) was investigated. Adding 30% giraffe, sloth bear or koala dung to the AnS inoculum yielded, respectively, a 11.17 (±4.51), 10.10 (±1.23), and 1.41 (±0.56)% higher biomethane production, as compared to the control (FW with solely AnS). The highest biomethane production of 564.00 (±3.88) ml CH 4 /gVS added obtained with 30% giraffe dung and 70% AnS was attributed to a higher solubilization of proteins (6.96 ± 2.76%) and recalcitrant carbohydrates (344.85 ± 54.31 mg/L as compared to zero). The biomethanation process could have been stimulated by the microorganisms or enzymes newly introduced, and/or the trace elements (Ni, Zn, and Co) present in the giraffe dung. These results indicate that bioaugmentation with zoo animals dung is worthy of further investigation as a strategy for improving the biomethane recovery from organic wastes.

Ehrlichia phagocytophila genogroup rickettsiae in ixodid ticks from California collected in 1995 and 1996.

PubMed Central

Barlough, J E; Madigan, J E; Kramer, V L; Clover, J R; Hui, L T; Webb, J P; Vredevoe, L K

1997-01-01

A total of 1,246 ixodid ticks collected in 1995 and 1996 from seven California counties were examined for the presence of Ehrlichia phagocytophila genogroup rickettsiae by using a nested PCR technique. Of 1,112 adult Ixodes pacificus Cooley and Kohls ticks tested, nine pools, each containing five ticks, were positive (minimum percentage of ticks harboring detectable ehrlichiae, 0.8%). Positive ticks were limited to four of the seven counties (Sonoma, El Dorado, Santa Cruz, and Orange). In Santa Cruz County, three positive pools were identified at the home of an individual with prior confirmed human granulocytic ehrlichiosis. In El Dorado County, positive ticks were found at sites where cases of granulocytic ehrlichiosis in a horse and a llama had recently occurred. Among 47 nymphal I. pacificus ticks collected in Sonoma County, one positive pool was identified. Fifty-seven adult Dermacentor occidentalis Marx and 30 adult D. variabilis Say ticks, collected chiefly in southern California, were negative. These data, although preliminary, suggest that the prevalence of E. phagocytophila genogroup rickettsiae in ixodid ticks of California may be lower than in cognate vector populations (i.e., I. scapularis Say = I. dammini Spielman, Clifford, Piesman, and Corwin) in the eastern and midwestern United States. PMID:9230373

Ehrlichia phagocytophila genogroup rickettsiae in ixodid ticks from California collected in 1995 and 1996.

PubMed

Barlough, J E; Madigan, J E; Kramer, V L; Clover, J R; Hui, L T; Webb, J P; Vredevoe, L K

1997-08-01

A total of 1,246 ixodid ticks collected in 1995 and 1996 from seven California counties were examined for the presence of Ehrlichia phagocytophila genogroup rickettsiae by using a nested PCR technique. Of 1,112 adult Ixodes pacificus Cooley and Kohls ticks tested, nine pools, each containing five ticks, were positive (minimum percentage of ticks harboring detectable ehrlichiae, 0.8%). Positive ticks were limited to four of the seven counties (Sonoma, El Dorado, Santa Cruz, and Orange). In Santa Cruz County, three positive pools were identified at the home of an individual with prior confirmed human granulocytic ehrlichiosis. In El Dorado County, positive ticks were found at sites where cases of granulocytic ehrlichiosis in a horse and a llama had recently occurred. Among 47 nymphal I. pacificus ticks collected in Sonoma County, one positive pool was identified. Fifty-seven adult Dermacentor occidentalis Marx and 30 adult D. variabilis Say ticks, collected chiefly in southern California, were negative. These data, although preliminary, suggest that the prevalence of E. phagocytophila genogroup rickettsiae in ixodid ticks of California may be lower than in cognate vector populations (i.e., I. scapularis Say = I. dammini Spielman, Clifford, Piesman, and Corwin) in the eastern and midwestern United States.

Development and Application of Camelid Molecular Cytogenetic Tools

PubMed Central

Avila, Felipe; Das, Pranab J.; Kutzler, Michelle; Owens, Elaine; Perelman, Polina; Rubes, Jiri; Hornak, Miroslav; Johnson, Warren E.

2014-01-01

Cytogenetic chromosome maps offer molecular tools for genome analysis and clinical cytogenetics and are of particular importance for species with difficult karyotypes, such as camelids (2n = 74). Building on the available human–camel zoo-fluorescence in situ hybridization (FISH) data, we developed the first cytogenetic map for the alpaca (Lama pacos, LPA) genome by isolating and identifying 151 alpaca bacterial artificial chromosome (BAC) clones corresponding to 44 specific genes. The genes were mapped by FISH to 31 alpaca autosomes and the sex chromosomes; 11 chromosomes had 2 markers, which were ordered by dual-color FISH. The STS gene mapped to Xpter/Ypter, demarcating the pseudoautosomal region, whereas no markers were assigned to chromosomes 14, 21, 22, 28, and 36. The chromosome-specific markers were applied in clinical cytogenetics to identify LPA20, the major histocompatibility complex (MHC)-carrying chromosome, as a part of an autosomal translocation in a sterile male llama (Lama glama, LGL; 2n = 73,XY). FISH with LPAX BACs and LPA36 paints, as well as comparative genomic hybridization, were also used to investigate the origin of the minute chromosome, an abnormally small LPA36 in infertile female alpacas. This collection of cytogenetically mapped markers represents a new tool for camelid clinical cytogenetics and has applications for the improvement of the alpaca genome map and sequence assembly. PMID:23109720

Isolation of Escherichia coli 0157:H7 Strain from Fecal Samples of Zoo Animal

PubMed Central

Mohammed Hamzah, Aseel; Mohammed Hussein, Aseel; Mahmoud Khalef, Jenan

2013-01-01

The isolation and characterization of Escherichia coli O157:H7 strains from 22 out of 174 fecal samples from petting zoo animals representing twenty-two different species (camel, lion, goats, zebra, bear, baboon monkey, Siberian monkey, deer, elk, llama, pony, horses, fox, kangaroo, wolf, porcupine, chickens, tiger, ostrich, hyena, dogs, and wildcats) were investigated. One petting Al-Zawraa zoological society of Baghdad was investigated for E. coli O157:H7 over a 16-month period that spanned two summer and two autumn seasons. Variation in the occurrence of E. coli O157:H7-positive petting zoo animals was observed, with animals being culture positive only in the summer months but not in the spring, autumn, or winter. E. coli O157:H7 isolates were distinguished by agglutination with E. coli O157:H7 latex reagent (Oxoid), identified among the isolates, which showed that multiple E. coli strains were isolated from one petting zoo animal, in which a single animal simultaneously shed multiple E. coli strains; E. coli O157:H7 was isolated only by selective enrichment culture of 2 g of petting zoo animal feces. In contrast, strains other than O157:H7 were cultured from feces of petting zoo animals without enrichment. PMID:24489514

Occurrence of gastrointestinal parasites in wild animals in State of Paraná, Brazil.

PubMed

Sprenger, Lew K; Yoshitani, Ursula Y; Buzatti, Andreia; Molento, Marcelo B

2018-01-01

The objective of this study was to determine the prevalence and diversity of gastrointestinal parasites in fecal samples from wild birds and mammals from the State of Paraná. In total, 220 stool samples were sent to Parasitic Diseases Laboratory of the Federal University of Paraná during 13 months (Jan/2013-Jan/2014). A total of 52.7% (116/220) of the animals were positive for cysts, oocysts, eggs and/or trophozoites. In birds, the positivity rate was 37.9% (25/66) and mammals was 59.1% (91/154). Strongyloidea superfamily eggs were observed in 37.3% (82/220) of the samples, Eimeria spp. in 10% (22/220), and Trichuris spp. in 4.5% (10/220). The most frequent mammal species were llamas (Lama glama), and dromedaries (Camelus bactrianus) with infection rate of 70.1% (54/77) and 60.8% (14/23), respectively. In other hand, cockatiels (Nymphicus hollandicus) and ring necks (Psittacula krameri), were the most researched birds, with infection rate of 20% (40/50) and 100% (6/6), respectively. A high prevalence of gastrointestinal parasites was observed in most of wildlife animals. Further investigations should be conducted focusing on parasite control strategies and the conservation measurements for harmonizing the human-animal interaction on the long-term, reducing associated health risks.

Improving solubility and refolding efficiency of human V(H)s by a novel mutational approach.

PubMed

Tanha, Jamshid; Nguyen, Thanh-Dung; Ng, Andy; Ryan, Shannon; Ni, Feng; Mackenzie, Roger

2006-11-01

The antibody V(H) domains of camelids tend to be soluble and to resist aggregation, in contrast to human V(H) domains. For immunotherapy, attempts have therefore been made to improve the properties of human V(H)s by camelization of a small set of framework residues. Here, we have identified through sequence comparison of well-folded llama V(H) domains an alternative set of residues (not typically camelid) for mutation. Thus, the solubility and thermal refolding efficiency of a typical human V(H), derived from the human antibody BT32/A6, were improved by introduction of two mutations in framework region (FR) 1 and 4 to generate BT32/A6.L1. Three more mutations in FR3 of BT32/A6.L1 further improved the thermal refolding efficiency while retaining solubility and cooperative melting profiles. To demonstrate practical utility, BT32/A6.L1 was used to construct a phage display library from which were isolated human V(H)s with good antigen binding activity and solubility. The engineered human V(H) domains described here may be useful for immunotherapy, due to their expected low immunogenicity, and in applications involving transient high temperatures, due to their efficient refolding after thermal denaturation.

Selection and expression of recombinant single domain antibodies from a hyper-immunized library against the hapten azoxystrobin.

PubMed

Makvandi-Nejad, Shokouh; Fjällman, Ted; Arbabi-Ghahroudi, Mehdi; MacKenzie, C Roger; Hall, J Christopher

2011-10-28

Three V(H)Hs against the model hapten, azoxystrobin (MW 403), were isolated from a hyper-immunized phage-displayed V(H)H library. This library was constructed by isolating the V(H)H-coding genes from the lymphocytes collected from a Llama glama that was immunized with azoxystrobin conjugated to bovine serum albumin (BSA). Six rounds of panning were performed against azoxystrobin conjugated to either ovalbumin (OVA) or rabbit serum albumin (RSA) to enrich clones containing V(H)Hs specific to the hapten. After screening 95 clones, three V(H)Hs (A27, A72, and A85) with different amino acid sequences were identified, expressed in soluble format in Escherichia coli HB2151, and purified using nickel-immobilized metal affinity chromatography. Competitive inhibition enzyme-linked immunosorbent assay (CI-ELISA) showed that A27 and A85 were specific to azoxystrobin while A72 was not. The IC(50) values of A27 and A85 V(H)Hs were 7.2 and 2.0μM, respectively. To our knowledge A85 is one of the highest affinity V(H)Hs that has yet been isolated against a hydrophobic hapten such as azoxystrobin. Copyright © 2011 Elsevier B.V. All rights reserved.

Single domain antibodies are specially suited for quantitative determination of gliadins under denaturing conditions.

PubMed

Doña, Vanina; Urrutia, Mariela; Bayardo, Mariela; Alzogaray, Vanina; Goldbaum, Fernando Alberto; Chirdo, Fernando G

2010-01-27

Food intended for celiac patients' consumption must be analyzed for the presence of toxic prolamins using high detectability tests. Though 60% ethanol is the most commonly used solvent for prolamins extraction, 2-mercaptoethanol (2-ME) and guanidinium chloride (GuHCl) can be added to increase protein recovery. However, ethanol and denaturing agents interfere with antigen recognition when conventional antibodies are used. In the present work, a new method for gliadins quantification is shown. The method is based on the selection of llama single domain antibody fragments able to operate under denaturing conditions. Six out of 28 VHH-phages obtained retained their binding capacity in 15% ethanol. Selected clones presented a long CDR3 region containing two additional cysteines that could be responsible for the higher stability. One of the clones (named VHH26) was fully operative in the presence of 15% ethanol, 0.5% 2-ME, and 0.5 M GuHCl. Capture ELISA using VHH26 was able to detect gliadins in samples shown as negatives by conventional ELISA. Therefore, this new strategy appears as an excellent platform for quantitative determination of proteins or any other immunogenic compound, in the presence of denaturing agents, when specific recognition units with high stability are required.

Isolation of Escherichia coli 0157:H7 strain from fecal samples of zoo animal.

PubMed

Mohammed Hamzah, Aseel; Mohammed Hussein, Aseel; Mahmoud Khalef, Jenan

2013-01-01

The isolation and characterization of Escherichia coli O157:H7 strains from 22 out of 174 fecal samples from petting zoo animals representing twenty-two different species (camel, lion, goats, zebra, bear, baboon monkey, Siberian monkey, deer, elk, llama, pony, horses, fox, kangaroo, wolf, porcupine, chickens, tiger, ostrich, hyena, dogs, and wildcats) were investigated. One petting Al-Zawraa zoological society of Baghdad was investigated for E. coli O157:H7 over a 16-month period that spanned two summer and two autumn seasons. Variation in the occurrence of E. coli O157:H7-positive petting zoo animals was observed, with animals being culture positive only in the summer months but not in the spring, autumn, or winter. E. coli O157:H7 isolates were distinguished by agglutination with E. coli O157:H7 latex reagent (Oxoid), identified among the isolates, which showed that multiple E. coli strains were isolated from one petting zoo animal, in which a single animal simultaneously shed multiple E. coli strains; E. coli O157:H7 was isolated only by selective enrichment culture of 2 g of petting zoo animal feces. In contrast, strains other than O157:H7 were cultured from feces of petting zoo animals without enrichment.

Atrial Fibrillation in Eight New World Camelids.

PubMed

Bozorgmanesh, R; Magdesian, K G; Estell, K E; Stern, J A; Swain, E A; Griffiths, L G

2016-01-01

There is limited information on the incidence of clinical signs, concurrent illness and treatment options for atrial fibrillation (AF) in New World Camelids (NWC). Describe clinical signs and outcome of AF in NWC. Eight New World Camelids admitted with AF. A retrospective observational study of camelids diagnosed with AF based on characteristic findings on electrocardiogram (ECG). All animals had an irregularly irregular heart rhythm detected on physical examination and 4 cases had obtunded mentation on admission. Three camelids were diagnosed with AF secondary to oleander intoxication, 3 animals had underlying cardiovascular disease, 1 was diagnosed with lone AF and 1 had AF diagnosed on examination for a urethral obstruction. Five of eight animals survived to discharge and nonsurvivors consisted of animals which died or were euthanized as a result of cardiovascular disease (2/8) or extra-cardiac disease unrelated to the AF (1/8). Atrial fibrillation occurs in NWC in association with cardiovascular disease, extra-cardiac disease or as lone AF. Amiodarone and transthoracic cardioversion were attempted in one llama with lone AF, but were unsuccessful. Atrial fibrillation was recorded in 0.1% of admissions. Copyright © 2015 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

[The structure of the gastric mucosa of the llamas (Lama guanocoe and Lama lamae). I. Forestomach].

PubMed

Luciano, L; Voss-Wermbter, G; Behnke, M; von Engelhardt, W; Reale, E

1979-01-01

The mucous membrane of the first and second compartments (ventral regions) as well as of the third compartment of Lama guanacoe and Lama lamae stomach shows tubular glands opening into pits. Below the surface epithelium blood capillaries of the fenestrated type form a regular network, each mesh of which surrounds a gastric pit. From a morphological point of view (thin section and freeze-fracture replicas) the columnar cells of the surface epithelium and those of the pits closest to the capillaries are largely similar to the epithelial cells of the rabbit gallbladder. This similarity suggests that at the level of the columnar cells sodium-dependent water reabsorption occurs. This reabsorption has already been demonstrated in the abovementioned compartments by physiological methods. The surface and foveolae epithelial cells as well as some cells of the tubular glands have a secretory function. Their secretory granules contain mucosubstances, as indicated by light-(PAS- and Alcian blue reactions) and electron microscopic (PA-TCH-Ag-reaction) histochemistry. The secretory granules originate from the Golgi complex which shows a positive histochemical reaction in its innermost sacculi at the electron microscope level. Endocrine cells (s. second part of this investigation) are rare. The mucosal membrane of each muscular lip separating the glandular sacs in the first compartment shows a stratified, not keratinized, squamous epithelium.

Llama VHH antibody fragments against GFAP: better diffusion in fixed tissues than classical monoclonal antibodies.

PubMed

Perruchini, Claire; Pecorari, Frederic; Bourgeois, Jean-Pierre; Duyckaerts, Charles; Rougeon, François; Lafaye, Pierre

2009-11-01

Camelids produce antibodies made of homodimeric heavy chains, and the antigen-binding region being composed of a single domain called VHH. These VHHs are much smaller than complete IgG. They are also more thermostable and more soluble in water; they should, therefore, diffuse more readily in the tissues. VHHs, expressed in bacteria, are easier to produce than conventional monoclonal antibodies. Because of these special characteristics, these antibody fragments could have interesting developments in immunohistochemistry and in the development of biomarkers. To test the possibility of their use in immunohistochemistry (IHC), we selected the glial fibrillary acidic protein (GFAP), a well-known marker of astrocytes. One alpaca (Lama pacos) was immunized against GFAP. Lymphocytes were isolated; the DNA was extracted; the VHH-coding sequences were selectively amplified. Three VHHs with a high affinity for GFAP and their corresponding mRNA were selected by ribosome display. Large quantities of the recombinant VHHs coupled with different tags were harvested from transfected bacteria. One of them was shown to immunolabel strongly and specifically to GFAP of human astrocytes in tissue sections. The quality of the IHC was comparable or, in some aspects, superior to the quality obtained with conventional IgG. The VHH was shown to diffuse on a longer distance than conventional monoclonal antibodies in fixed cortical tissue: a property that may be useful in immunolabeling of thick sections.

QUBIC in Argentina

NASA Astrophysics Data System (ADS)

García, B.; Harari, D.; Etchegoyen, A.; Medina, M. C.; Romero, G. E.; Qubic Collaboration

2017-10-01

QUBIC (QU Bolometric Interferometer for Cosmology) is an experimental cosmology project to measure the modes in the polarization of the cosmic background radiation (CBR), the fossil relic that reveals the properties at the Universe 380,000 years after the Planck Era. Discovery of the CBR in 1964 and measurement of tiny temperature anisotropies in 1992 were major breakthroughs in our understanding of the Universe. The next challenge is to measure the polarization of the CBR accurately enough to detect modes that would reveal the existence of primordial gravitational waves produced in the first stages of the Planck Era and probe inflation theory, that assumes an accelerated expansion during the first seconds. The mode signal is however extremely weak and its measurement requires complex instruments. The QUBIC collaboration has developed the concept of interferometric bolometry, that brings together the sensitivity of bolometric detectors with the control of systematic effects provided by interferometry. QUBIC is an international collaboration involving several universities and laboratories in France, Italy, United Kingdom and USA. Recently, Argentina has suggested Alto Chorrillo (Salta), as candidate site for the installation of the experiment, next to the LLAMA site. Here we describe the scientific objectives and the main features of the experiment and we detail the process through which the international collaboration decided to install in Argentina its first module as well as the challenges for our country in this project.

Market access and community size influence pastoral management of native and exotic livestock species: A case study in communities of the Cordillera Real in Bolivia's high Andean wetlands.

PubMed

Struelens, Quentin; Gonzales Pomar, Karina; Loza Herrera, Susi; Nina Huanca, Gaby; Dangles, Olivier; Rebaudo, François

2017-01-01

Grazing areas management is of utmost importance in the Andean region. In the valleys of the Bolivian Cordillera Real near La Paz, pastoralism constitutes the traditional way for people to insure food security and economical sustainability. In these harsh mountains, unique and productive wetlands sustained by glacial water streams are of utmost importance for feeding cattle herds during the dry season. After the colonization by the Spanish, a shift in livestock species has been observed, with the introduction of exotic species such as cows and sheep, resulting in a different impact on pastures compared to native camelid species-llamas and alpacas. Here we explored some of the social-economical and environmental drivers that motivate Bolivian pastoralists to prefer exotic over native livestock species, based on 36 household surveys in the Cordillera Real. We constructed a Partial Least Squares Structural Equation Model in order to assess the relationships between these drivers. Our results suggest that the access to market influenced pastoralists to reshape their herd composition, by increasing the number of sheep. They also suggest that community size increased daily grazing time in pastures, therefore intensifying the grazing pressure. At a broader scale, this study highlights the effects of some social-economical and environmental drivers on mountain herding systems.

Market access and community size influence pastoral management of native and exotic livestock species: A case study in communities of the Cordillera Real in Bolivia's high Andean wetlands

PubMed Central

Struelens, Quentin; Gonzales Pomar, Karina; Loza Herrera, Susi; Nina Huanca, Gaby; Dangles, Olivier

2017-01-01

Grazing areas management is of utmost importance in the Andean region. In the valleys of the Bolivian Cordillera Real near La Paz, pastoralism constitutes the traditional way for people to insure food security and economical sustainability. In these harsh mountains, unique and productive wetlands sustained by glacial water streams are of utmost importance for feeding cattle herds during the dry season. After the colonization by the Spanish, a shift in livestock species has been observed, with the introduction of exotic species such as cows and sheep, resulting in a different impact on pastures compared to native camelid species—llamas and alpacas. Here we explored some of the social-economical and environmental drivers that motivate Bolivian pastoralists to prefer exotic over native livestock species, based on 36 household surveys in the Cordillera Real. We constructed a Partial Least Squares Structural Equation Model in order to assess the relationships between these drivers. Our results suggest that the access to market influenced pastoralists to reshape their herd composition, by increasing the number of sheep. They also suggest that community size increased daily grazing time in pastures, therefore intensifying the grazing pressure. At a broader scale, this study highlights the effects of some social-economical and environmental drivers on mountain herding systems. PMID:29228062

Current knowledge and future challenges in camelid reproduction.

PubMed

Tibary, A; Anouassi, A; Sghiri, A; Khatir, H

2007-01-01

Reproductive biology research on camelids offers some interesting peculiarities and challenges to scientists and animal production specialists. The objective of this paper is to review camelid reproduction, advances in reproductive physiology and reproductive biotechnologies in camelids and discuss some areas for further research. In the female, the focus has been on understanding follicular dynamics. This has allowed development of synchronization and superovulation strategies to support embryo transfer technologies which are now commonly used in camels. Some advances have been achieved in preservation of embryos by vitrification. Fertilization, early embryo development and embryo signaling for maternal recognition of pregnancy are still not fully understood. New information on the interaction of the developing embryo and the endometrium may shed some light on this signaling as well as the mechanism of prevention of luteolysis. The presence of a seminal ovulation-inducing factor (OIF) was confirmed in llamas and alpacas. Chronology of oocytes maturation has been described. In vitro production of embryos has been achieved resulting in successful pregnancies and births in the dromedary. These techniques offer a new tool for the production and study of interspecies/cross-species embryos and their effect on pregnancy. Male reproductive function remains poorly studied. Semen preservation and artificial insemination still present many challenges and are not used in production at the moment. The involvement of climatic and nutritional conditions as well as the role of leptin in the regulation of reproductive function need to be evaluated.

Evaluation of a postexposure rabies prophylaxis protocol for domestic animals in Texas: 2000-2009.

PubMed

Wilson, Pamela J; Oertli, Ernest H; Hunt, Patrick R; Sidwa, Thomas J

2010-12-15

To determine whether postexposure rabies prophylaxis (PEP) in domestic animals, as mandated in Texas, has continued to be effective and to evaluate preexposure or postexposure vaccination failures from 2000 through 2009. Retrospective case series. 1,014 unvaccinated domestic animals (769 dogs, 126 cats, 72 horses, 39 cattle, 3 sheep, 4 goats, and 1 llama) that received PEP and 12 vaccinated domestic animals (7 dogs and 5 cats) with possible failure of protection. Zoonotic incident reports from 2000 through 2009 were reviewed for information regarding unvaccinated domestic animals that received PEP in accordance with the state protocol after exposure to a laboratory-confirmed rabid animal; reports also were reviewed for any preexposure or postexposure vaccination failures. The state-required PEP protocol was as follows: immediately vaccinate the animal against rabies, isolate the animal for 90 days, and administer booster vaccinations during the third and eighth weeks of the isolation period. From 2000 through 2009, 1,014 animals received PEP; no failures were recorded. One preexposure vaccination failure was recorded. The Texas PEP protocol was used during the 10-year period. Results indicated that an effective PEP protocol for unvaccinated domestic animals exposed to rabies was immediate vaccination against rabies, a strict isolation period of 90 days, and administration of booster vaccinations during the third and eighth weeks of the isolation period.

Dystrophic mineralization of the arterial fibrovascular tissue associated with a vitamin D hypervitaminosis in an 8-year-old female Alpaca (Vicugna pacos).

PubMed

Tavella, Alexander; Stefani, Annalisa; Zanardello, Claudia; Bettini, Astrid; Gauly, Matthias; Zanolari, Patrik

2016-01-01

Prophylactic Vitamin D supplementation is a common practice in Alpaca breeding in many regions around the world. An overdosage can lead to dystrophic mineralization of soft tissues. In this paper we illustrate a suspected case of hypervitaminosis D in an 8-year-old female Alpaca. In June 2015, the carcass of an 8-year-old female Alpaca ( Vicugna pacos ) was submitted to the diagnostic laboratory of the Istituto Zooprofilattico Sperimentale delle Venezie (IZSVe) for necropsy. The animal had undergone a spontaneous abortion with uterine prolapse and delivery of the placenta, and had died shortly thereafter. Death occurred due to internal haemorrhage related to dystrophic mineralization of the left renal artery with subsequent rupture and damage of the renal hilum. During the necropsy, histopathological and serum biochemical analyses were performed. After laboratory analyses and the history of mineral and vitamin supplementation reported by the breeder, a hypervitaminosis D was suspected to be the most probable cause of the dystrophic mineralization observed in the left renal artery. Most of the information regarding Llamas and Alpacas comes from the South American and Australian regions. It is therefore important to provide scientific information about these animals in other regions of the world in order to have a better and wider understanding of the nutritional and environmental conditions necessary for optimal breeding.

Mycobacterium bovis infections in domesticated non-bovine mammalian species. Part 1: Review of epidemiology and laboratory submissions in Great Britain 2004-2010.

PubMed

Broughan, J M; Downs, S H; Crawshaw, T R; Upton, P A; Brewer, J; Clifton-Hadley, R S

2013-11-01

Mycobacterium bovis, the causative agent of bovine tuberculosis (bTB), can infect a broad range of mammalian species in addition to domestic and feral cattle and badgers. Since legislation introduced in 2006 in Great Britain requires animal keepers, meat inspectors and veterinarians to notify the authorities of suspect bTB lesions or the isolation of M. bovis in any mammal excluding humans, the organism has been increasingly identified in domestic species other than cattle. Although in most cases 'spill-over' hosts, these remain a potential source of infection for cattle, wildlife, and possibly humans. In this first part of a two-part review of M. bovis infections in non-bovine domestic species, current knowledge of the epidemiology of such infections is presented along with novel data relating to diagnostic submissions for mycobacterial culture between 2004 and 2010. Over this period M. bovis infection was identified in 116 cats, 7 dogs, 34 llamas, 133 alpacas, 35 goats, 24 sheep and 85 pigs and wild boar. The risk that such infections pose to the control of bTB, and as zoonoses, is discussed. In part two, the options available to diagnose bTB in these species, as well as the challenges posed to disease detection and control will be discussed in depth. Copyright © 2013. Published by Elsevier Ltd.

Preclinical Development of a Novel, Orally-Administered Anti-Tumour Necrosis Factor Domain Antibody for the Treatment of Inflammatory Bowel Disease.

PubMed

Crowe, J Scott; Roberts, Kevin J; Carlton, Timothy M; Maggiore, Luana; Cubitt, Marion F; Clare, Simon; Harcourt, Katherine; Reckless, Jill; MacDonald, Thomas T; Ray, Keith P; Vossenkämper, Anna; West, Michael R

2018-03-21

TNFα is an important cytokine in inflammatory bowel disease. V565 is a novel anti-TNFα domain antibody developed for oral administration in IBD patients, derived from a llama domain antibody and engineered to enhance intestinal protease resistance. V565 activity was evaluated in TNFα-TNFα receptor-binding ELISAs as well as TNFα responsive cellular assays and demonstrated neutralisation of both soluble and membrane TNFα with potencies similar to those of adalimumab. Although sensitive to pepsin, V565 retained activity after lengthy incubations with trypsin, chymotrypsin, and pancreatin, as well as mouse small intestinal and human ileal and faecal supernatants. In orally dosed naïve and DSS colitis mice, high V565 concentrations were observed in intestinal contents and faeces and immunostaining revealed V565 localisation in mouse colon tissue. V565 was detected by ELISA in post-dose serum of colitis mice, but not naïve mice, demonstrating penetration of disrupted epithelium. In an ex vivo human IBD tissue culture model, V565 inhibition of tissue phosphoprotein levels and production of inflammatory cytokine biomarkers was similar to infliximab, demonstrating efficacy when present at the disease site. Taken together, results of these studies provide confidence that oral V565 dosing will be therapeutic in IBD patients where the mucosal epithelial barrier is compromised.

Creating wealth from groundwater for dollar-a-day farmers: Where the silent revolution and the four revolutions to end rural poverty meet

NASA Astrophysics Data System (ADS)

Polak, Paul; Yoder, Robert

2006-03-01

More than 550 million of the current 1.1 billion people earning less than 1-a-day earn a living from agriculture in developing countries. A revolution in water control is needed to develop and mass-disseminate new, affordable, small-plot irrigation technologies. A revolution in agriculture is required to enable smallholders to produce high-value, marketable, labor-intensive cash crops. A revolution in markets is needed to open access to markets for the crops they produce and the inputs they need to produce them. Finally, a revolution in design, based on the ruthless pursuit of affordability, is needed to harness shallow groundwater. The experiences of suppliers of treadle pumps, low-cost drip irrigation and water storage systems were examined. The wealth these technologies generated, coupled with falling prices for small diesel pumps in countries like India and China, created a suitable environment for the rapid adoption of affordable diesel pump tubewells, which in turn created vigorous water markets and expanded access to affordable irrigation water for smallholders. The combination of smallholder-centered revolutions, along with the ‘silent revolution in groundwater’ described by Llamas and Martinez-Santos (Water Sci Technol 51(8):167-174, 2005) provide new practical options for meeting the UN Millennium Development Goals on poverty and hunger by 2015.

Characterization of single-domain antibodies with an engineered disulfide bond.

PubMed

Hussack, Greg; Mackenzie, C Roger; Tanha, Jamshid

2012-01-01

Camelidae single-domain antibodies (VHHs) represent a unique class of emerging therapeutics. Similar to other recombinant antibody fragments (e.g., Fabs, scFvs), VHHs are amenable to library screening and selection, but benefit from superior intrinsic biophysical properties such as high refolding efficiency, high solubility, no tendency for aggregation, resistance to proteases and chemical denaturants, and high expression, making them ideal agents for antibody-based drug design. Despite these favorable biophysical characteristics, further improvements to VHH stability are desirable when considering applications in adverse environments like high heat, low humidity, pH extremes, and the acidic, protease-rich gastrointestinal tract. Recently, the introduction of a disulfide bond into the hydrophobic core of camelid VHHs increased antibody thermal and conformational stability. Here, we present additional protocols for characterizing the effects of the introduced disulfide bond on a panel of llama VHHs. Specifically, we employ mass spectrometry fingerprinting analysis of VHH peptides to confirm the presence of the introduced disulfide bond, size exclusion chromatography, and surface plasmon resonance to examine the effects on aggregation state and target affinity, and circular dichroism spectroscopy and protease digestion assays to assess the effects on thermal and proteolytic stability. The disulfide bond stabilization strategy can be incorporated into antibody library design and should lead to hyperstabilized single-domain antibodies (VHHs, VHs), and possibly Fabs and scFvs, if selection pressures such as denaturants or proteases are introduced during antibody selection.

Rugged Single Domain Antibody Detection Elements for Bacillus anthracis Spores and Vegetative Cells

PubMed Central

Walper, Scott A.; Anderson, George P.; Brozozog Lee, P. Audrey; Glaven, Richard H.; Liu, Jinny L.; Bernstein, Rachel D.; Zabetakis, Dan; Johnson, Linwood; Czarnecki, Jill M.; Goldman, Ellen R.

2012-01-01

Significant efforts to develop both laboratory and field-based detection assays for an array of potential biological threats started well before the anthrax attacks of 2001 and have continued with renewed urgency following. While numerous assays and methods have been explored that are suitable for laboratory utilization, detection in the field is often complicated by requirements for functionality in austere environments, where limited cold-chain facilities exist. In an effort to overcome these assay limitations for Bacillus anthracis, one of the most recognizable threats, a series of single domain antibodies (sdAbs) were isolated from a phage display library prepared from immunized llamas. Characterization of target specificity, affinity, and thermal stability was conducted for six sdAb families isolated from rounds of selection against the bacterial spore. The protein target for all six sdAb families was determined to be the S-layer protein EA1, which is present in both vegetative cells and bacterial spores. All of the sdAbs examined exhibited a high degree of specificity for the target bacterium and its spore, with affinities in the nanomolar range, and the ability to refold into functional antigen-binding molecules following several rounds of thermal denaturation and refolding. This research demonstrates the capabilities of these sdAbs and their potential for integration into current and developing assays and biosensors. PMID:22412927

Changing Hydrology in Glacier-fed High Altitude Andean Peatbogs

NASA Astrophysics Data System (ADS)

Slayback, D. A.; Yager, K.; Baraer, M.; Mohr, K. I.; Argollo, J.; Wigmore, O.; Meneses, R. I.; Mark, B. G.

2012-12-01

Montane peatbogs in the glacierized Andean highlands of Peru and Bolivia provide critical forage for camelids (llama and alpaca) in regionally extensive pastoral agriculture systems. During the long dry season, these wetlands often provide the only available green forage. A key question for the future of these peatbog systems, and the livelihoods they support, is the impact of climate change and glacier recession on their hydrology, and thus forage production. We have already documented substantial regional glacier recession, of, on average, approximately 30% of surface area over the past two decades. As glaciers begin to retreat under climate change, there is initially a period of increased meltwater outflow, culminating in a period of "peak water", and followed by a continual decline in outflows. Based on previous work, we know that some glaciers in the region have already passed peak water conditions, and are now declining. To better understand the impacts of these processes on peatbog hydrology and productivity, we have begun collecting a variety of surface data at several study sites in both Bolivia and Peru. These include precipitation, stream flow, water levels, water chemistry and isotope analyses, and peatbog biodiversity and biomass. These measurements will be used in conjunction with a regional model driven by satellite data to predict likely future impacts. We will present the results from these initial surface measurements, and an overview of satellite datasets to be used in the regional model.

Evaluation of a panel of antibodies for the immunohistochemical identification of immune cells in paraffin-embedded lymphoid tissues of new- and old-world camelids.

PubMed

Uhde, Ann-Kathrin; Lehmbecker, Annika; Baumgärtner, Wolfgang; Spitzbarth, Ingo

2017-02-01

Different species of camelids play an important role in the epidemiology of various emerging infectious diseases such as Middle East respiratory syndrome. For precise investigations of the immunopathogenesis in these host species, appropriate immunohistochemical markers are highly needed in order to phenotype distinct immune cells populations in camelids. So far, specific immunohistochemical markers for camelid immune cells are rarely commercially available, and cross-reactivity studies are restricted to the use of frozen dromedary tissues. To bridge this gap, 14 commercially available primary antibodies were tested for their suitability to demonstrate immune cell populations on formalin fixed paraffin-embedded (FFPE) tissue sections of dromedaries, Bactrian camels, llamas, and alpacas in the present study. Out of these, 9 antibodies directed against CD3, CD20, CD79α, HLA-DR, Iba-1, myeloid/histiocyte antigen, CD204, CD208, and CD68 antigen exhibited distinct immunoreaction patterns to certain camelid immune cell subsets. The distribution of these antigens was comparatively evaluated in different anatomical compartments of thymus, spleen, mesenteric, and tracheobronchial lymph nodes. The presented results will provide a basis for further investigations in camelids, especially with respect to the role of the immune response in certain infectious diseases, which harbor a considerable risk to spill over to other species. Copyright © 2017 Elsevier B.V. All rights reserved.

A candidate gene for choanal atresia in alpaca.

PubMed

Reed, Kent M; Bauer, Miranda M; Mendoza, Kristelle M; Armién, Aníbal G

2010-03-01

Choanal atresia (CA) is a common nasal craniofacial malformation in New World domestic camelids (alpaca and llama). CA results from abnormal development of the nasal passages and is especially debilitating to newborn crias. CA in camelids shares many of the clinical manifestations of a similar condition in humans (CHARGE syndrome). Herein we report on the regulatory gene CHD7 of alpaca, whose homologue in humans is most frequently associated with CHARGE. Sequence of the CHD7 coding region was obtained from a non-affected cria. The complete coding region was 9003 bp, corresponding to a translated amino acid sequence of 3000 aa. Additional genomic sequences corresponding to a significant portion of the CHD7 gene were identified and assembled from the 2x alpaca whole genome sequence, providing confirmatory sequence for much of the CHD7 coding region. The alpaca CHD7 mRNA sequence was 97.9% similar to the human sequence, with the greatest sequence difference being an insertion in exon 38 that results in a polyalanine repeat (A12). Polymorphism in this repeat was tested for association with CA in alpaca by cloning and sequencing the repeat from both affected and non-affected individuals. Variation in length of the poly-A repeat was not associated with CA. Complete sequencing of the CHD7 gene will be necessary to determine whether other mutations in CHD7 are the cause of CA in camelids.

Streamlined method for parallel identification of single domain antibodies to membrane receptors on whole cells

PubMed Central

Rossotti, Martín; Tabares, Sofía; Alfaya, Lucía; Leizagoyen, Carmen; Moron, Gabriel; González-Sapienza, Gualberto

2015-01-01

BACKGROUND Owing to their minimal size, high production yield, versatility and robustness, the recombinant variable domain (nanobody) of camelid single chain antibodies are valued affinity reagents for research, diagnostic, and therapeutic applications. While their preparation against purified antigens is straightforward, the generation of nanobodies to difficult targets such as multi-pass or complex membrane cell receptors remains challenging. Here we devised a platform for high throughput identification of nanobodies to cell receptor based on the use of a biotin handle. METHODS Using a biotin-acceptor peptide tag, the in vivo biotinylation of nanobodies in 96 well culture blocks was optimized allowing their parallel analysis by flow cytometry and ELISA, and their direct used for pull-down/MS target identification. RESULTS The potential of this strategy was demonstrated by the selection and characterization of panels of nanobodies to Mac-1 (CD11b/CD18), MHC II and the mouse Ly-5 leukocyte common antigen (CD45) receptors, from a VHH library obtained from a llama immunized with mouse bone marrow derived dendritic cells. By on and off switching of the addition of biotin, the method also allowed the epitope binning of the selected Nbs directly on cells. CONCLUSIONS This strategy streamline the selection of potent nanobodies to complex antigens, and the selected nanobodies constitute ready-to-use biotinylated reagents. GENERAL SIGNIFICANCE This method will accelerate the discovery of nanobodies to cell membrane receptors which comprise the largest group of drug and analytical targets. PMID:25819371

Production, Preservation, and Transfer of South American Camelid Embryos

PubMed Central

Trasorras, Virginia L.; Carretero, María Ignacia; Neild, Deborah M.; Chaves, Maria Graciela; Giuliano, Susana M.; Miragaya, Marcelo H.

2017-01-01

The current review summarizes progress in the field of in vitro and in vivo production of South American Camelid embryos. Both methods require ovarian superstimulation (with FSH and eCG) to obtain multiple ovulations (in vivo embryo production) or to induce follicle growth for oocyte collection (in vitro embryo production). Moreover, superstimulation entails prior administration of hormones that inhibit follicular growth (progesterone, progestagens, and estrogens). Cumulus-oocyte complexes obtained must mature in vivo (buserelin administration) or in vitro to then be subjected to in vitro fertilization or intracytoplasmic sperm injection. All these techniques also require morphologically normal, motile spermatozoa to achieve fertilization. Methods used to decrease semen viscosity and to select the best spermatozoa (Percoll®; Androcoll-ETM) are described. Additionally, nuclear transfer or cloning has been applied in llamas. Up to now, embryo deep-freezing and vitrification have progressed slowly but are at the height of development. Embryos that are obtained by any of these techniques, either in vivo or in vitro, need to be transferred to synchronized recipient females. The best results are achieved after transfer to the left uterine horn with an ipsilateral ovulation. No live offspring have been obtained after the transfer of cryopreserved embryos. Applying reproductive biotechnologies, such as those described, will permit the expansion of genetically selected animals in the population and also that of wild camelid species, vicunas, and guanacos, whose embryos could then be transferred to the uterus of domestic species. PMID:29181380

Coronary Artery Anomalies in Animals

PubMed Central

Scansen, Brian A.

2017-01-01

Coronary artery anomalies represent a disease spectrum from incidental to life-threatening. Anomalies of coronary artery origin and course are well-recognized in human medicine, but have received limited attention in veterinary medicine. Coronary artery anomalies are best described in the dog, hamster, and cow though reports also exist in the horse and pig. The most well-known anomaly in veterinary medicine is anomalous coronary artery origin with a prepulmonary course in dogs, which limits treatment of pulmonary valve stenosis. A categorization scheme for coronary artery anomalies in animals is suggested, dividing these anomalies into those of major or minor clinical significance. A review of coronary artery development, anatomy, and reported anomalies in domesticated species is provided and four novel canine examples of anomalous coronary artery origin are described: an English bulldog with single left coronary ostium and a retroaortic right coronary artery; an English bulldog with single right coronary ostium and transseptal left coronary artery; an English bulldog with single right coronary ostium and absent left coronary artery with a prepulmonary paraconal interventricular branch and an interarterial circumflex branch; and a mixed-breed dog with tetralogy of Fallot and anomalous origin of all coronary branches from the brachiocephalic trunk. Coronary arterial fistulae are also described including a coronary cameral fistula in a llama cria and an English bulldog with coronary artery aneurysm and anomalous shunting vessels from the right coronary artery to the pulmonary trunk. These examples are provided with the intent to raise awareness and improve understanding of such defects. PMID:29056679

Humoral response to 2 inactivated bluetongue virus serotype-8 vaccines in South American camelids.

PubMed

Zanolari, P; Bruckner, L; Fricker, R; Kaufmann, C; Mudry, M; Griot, C; Meylan, M

2010-01-01

Bluetongue virus serotype 8 (BTV-8) has caused disease in domestic ruminants in several countries of northern Europe since 2006. In 2008 a mass-vaccination program was launched in most affected countries using whole virus inactivated vaccines. To evaluate 2 inactivated vaccines (Bovilis BTV 8; BTVPUR AlSap8) for immunogenicity and safety against BTV-8 in South American camelids (SAC) in a field trial. Forty-two SAC (25 Alpacas, 17 Llamas) aged between 1 and 16 years. The animals were vaccinated twice at intervals of 21 days. They were observed clinically for adverse local, systemic, or both reactions throughout the trial. Blood samples collected on days 0, 14, 21, 43, and 156 after vaccination were tested for the presence of BTV-8 virus by real time-polymerase chain reaction and of specific antibodies by competitive ELISA and a serum neutralization test. All vaccinated animals developed antibodies to BTV-8 after the 2nd administration of the vaccine. No adverse effects were observed except for moderate local swellings at the injection site, which disappeared within 21 days. Slightly increased body temperatures were only observed in the first 2 days after vaccination. The BTV was not detected in any of the samples analyzed. The administration of the 2 inactivated commercial vaccines was safe and induced seroconversion against BTV-8 in all vaccinated animals. The results of this study suggest that 2 doses injected 3 weeks apart is a suitable vaccination regimen for SAC.

Experimental infection of South American camelids with bluetongue virus serotype 8.

PubMed

Schulz, Claudia; Eschbaumer, Michael; Rudolf, Miriam; König, Patricia; Keller, Markus; Bauer, Christian; Gauly, Matthias; Grevelding, Christoph G; Beer, Martin; Hoffmann, Bernd

2012-01-27

Bluetongue (BT) is an infectious, non-contagious disease of wild and domestic ruminants. It is caused by bluetongue virus (BTV) and transmitted by Culicoides biting midges. Since 1998, BT has been emerging throughout Europe, threatening not only the naïve ruminant population. Historically, South American camelids (SAC) were considered to be resistant to BT disease. However, recent fatalities related to BTV in captive SAC have raised questions about their role in BTV epidemiology. Data on the susceptibility of SAC to experimental infection with BTV serotype 8 (BTV-8) were collected in an animal experiment. Three alpacas (Vicugna pacos) and three llamas (Lama glama) were experimentally infected with BTV-8. They displayed very mild clinical signs. Seroconversion was first measured 6-8 days after infection (dpi) by ELISA, and neutralising antibodies appeared 10-13 dpi. BTV-8 RNA levels in blood were very low, and quickly cleared after seroconversion. However, spleens collected post-mortem were still positive for BTV RNA, over 71 days after the last detection in blood samples. Virus isolation was only possible from blood samples of two alpacas by inoculation of highly sensitive interferon alpha/beta receptor-deficient (IFNAR(-/-)) mice. An in vitro experiment demonstrated that significantly lower amounts of BTV-8 adsorb to SAC blood cells than to bovine blood cells. Although this experiment showed that SAC are generally susceptible to a BTV-8 infection, it indicates that these species play a negligible role in BTV epidemiology. Copyright © 2011 Elsevier B.V. All rights reserved.

Prevalence and significance of gastrointestinal helminths and protozoa in South American Camelids in Switzerland.

PubMed

Hertzberg, Hubertus; Kohler, Lucia

2006-01-01

A cross sectional study was conducted to determine the prevalence and significance of endoparasitic infections in South American Camelids (SAC) in Switzerland. Qualitative and quantitative coproscopic examinations were performed in 38 farms during the grazing period. Management practices with possible interference with parasitic infections were analyzed. On the farm level prevalences of endoparasitic infections were: trichostrongyles 87%; Trichuris sp. 74%; Capillaria sp. 68%; Nematodirus battus 63%; Nematodirus sp. 53%; Dicrocoelium dendriticum 34%; Moniezia sp. 8%; Fasciola hepatica 5%; protostrongylids 5%; Eimeria macusaniensis 68%. The level of helminth egg excretion was generally low. The highest values were recorded for trichostrongyles with an average of all investigated farms of 53 eggs per gram of faeces. The mean trichostrongyle egg output was approximately three-fold in SAC on farms that also kept sheep and/or goats, although this difference was not significant (P = 0.11). Clinical trichostrongylidosis was not reported from any of the farms. The low infection level with gastrointestinal nematodes is attributed to the defaecation behaviour of the SAC depositing their faeces focally on small spots on pasture. As a consequence, pasture infectivity is largely restricted to the area adjacent to the dung piles. Dicrocoeliosis is regarded as the most relevant parasitic infection of llamas and alpacas in Switzerland causing severe clinical symptoms and death in untreated animals. Sixteen per cent of the owners regularily treated their herds against dicrocoeliosis using praziquantel at a dose of 50 mg/kg body weight orally.

Oriented conjugates of monoclonal and single-domain antibodies with quantum dots for flow cytometry and immunohistochemistry diagnostic applications

NASA Astrophysics Data System (ADS)

Sukhanova, Alyona; Even-Desrumeaux, Klervi; Millot, Jean-Marc; Chames, Patrick; Baty, Daniel; Artemyev, Mikhail; Oleinikov, Vladimir; Cohen, Jacques H. M.; Nabiev, Igor

2012-03-01

Ideal diagnostic nanoprobes should not exceed 15 nm in size and should contain high-affinity homogeneously oriented capture molecules on their surface. An advanced procedure for antibody (Ab) reduction was used to cleave each Ab into two functional half-Abs, 75-kDa heavy-light chain fragments, each containing an intact antigen-binding site. Affinity purification of half-Abs followed by their linkage to quantum dots (QDs) yielded oriented QD-Ab conjugates whose functionality was considerably improved compared to those obtained using the standard protocols. Ultrasmall diagnostic nanoprobes were engineered through oriented conjugation of QDs with 13-kDa single-domain Abs (sdAbs) derived from llama IgG. sdAbs were tagged with QDs via an additional cysteine residue specifically integrated into the C-terminal region of sdAb using genetic engineering. This approach made it possible to obtain sdAb-QD nanoprobes <12 nm in diameter comprising four copies of sdAbs linked to the same QD in an oriented manner. sdAb-QD conjugates against carcinoembryonic antigen (CEA) and HER2 exhibited an extremely high specificity in flow cytometry; the quality of immunohistochemical labeling of biopsy samples was found to be superior to that of labeling according to the current "gold standard" protocols of anatomo-pathological practice. The nano-bioengineering approaches developed can be extended to oriented conjugation of Abs and sdAbs with different semiconductor, noble metal, or magnetic nanoparticles.

Successful use of camelid (alpaca) antivenom to treat a potentially lethal tiger snake (Notechis scutatus) envenomation in a dog.

PubMed

Padula, Andrew M; Winkel, Kenneth D

2016-05-01

This report describes a confirmed clinical case of tiger snake (Notechis scutatus) envenomation in a domestic dog that was successfully treated with a novel polyvalent camelid (alpaca; Llama pacos) antivenom. Samples collected from the dog were assayed for tiger snake venom (TSV) using a highly sensitive and specific ELISA. The TSV concentration in serum and urine at initial presentation was 365 ng/mL and 11,640 ng/mL respectively. At the time of initial presentation whole blood collected from the dog did not clot and the Prothrombin Time was abnormally increased (>300 s). Serum was also visibly hemolysed. The dog was administered antihistamine, dexamethasone and 4000 Units (sufficient to neutralise 40 mg of TSV) of a novel polyvalent alpaca antivenom diluted in 0.9% NaCl. At 4 h post-antivenom treatment the dog's clinical condition had improved markedly with serum TSV concentrations below the limit of detection (<0.015 ng/mL), consistent with complete binding of venom antigens by the alpaca antivenom. Coagulation parameters had begun to improve by 4 h and had fully normalised by 16 h post-antivenom. Venom concentrations in both serum and urine remained undetectable at 16 h post-antivenom. The dog made a complete recovery, without complications, suggesting that the alpaca-based antivenom is both clinically safe and effective. Copyright © 2016 Elsevier Ltd. All rights reserved.

Single domain antibody-quantum dot conjugates for ricin detection by both fluoroimmunoassay and surface plasmon resonance.

PubMed

Anderson, George P; Glaven, Richard H; Algar, W Russ; Susumu, Kimihiro; Stewart, Michael H; Medintz, Igor L; Goldman, Ellen R

2013-07-05

The combination of stable biorecognition elements and robust quantum dots (QDs) has the potential to yield highly effective reporters for bioanalyses. Llama-derived single domain antibodies (sdAb) provide small thermostable recognition elements that can be easily manipulated using standard DNA methods. The sdAb was self-assembled on dihydrolipoic acid (DHLA) ligand-capped CdSe-ZnS core-shell QDs made in our laboratory through the polyhistidine tail of the protein, which coordinated to zinc ions on the QD surface. The sdAb-QD bioconjugates were then applied in both fluorometric and surface plasmon resonance (SPR) immunoassays for the detection of ricin, a potential biothreat agent. The sdAb-QD conjugates functioned in fluoroimmunoassays for the detection of ricin, providing equivalent limits of detection when compared to the same anti-ricin sdAb labeled with a conventional fluorophore. In addition, the DHLA-QD-sdAb conjugates were very effective reporter elements in SPR sandwich assays, providing more sensitive detection with a signal enhancement of ~10-fold over sdAb reporters and 2-4 fold over full sized antibody reporters. Commercially prepared streptavidin-modified polymer-coated QDs also amplified the SPR signal for the detection of ricin when applied to locations where biotinylated anti-ricin sdAb was bound to target; however, we observed a 4-fold greater amplification when using the DHLA-QD-sdAb conjugates in this format. Copyright © 2013. Published by Elsevier B.V.

Anaplasma infection of Bactrian camels (Camelus bactrianus) and ticks in Xinjiang, China.

PubMed

Li, Youquan; Yang, Jifei; Chen, Ze; Qin, Gege; Li, Yaqiong; Li, Qian; Liu, Junlong; Liu, Zhijie; Guan, Guiquan; Yin, Hong; Luo, Jianxun; Zhang, Lin

2015-06-10

To date, anaplasmosis has been reported to be a subclinical disease in Indian and Arabian one-humped camels (Camelus dromedarius) and llamas (Lama glama). However, no information on Anaplasma infection in two-humped Bactrian camels (Camelus bactrianus) in China has been published to date. The aim of this study was to investigate the prevalence of Anaplasma spp. in domestic Bactrian camels and ticks in Xinjiang, China. A total of 382 ticks were collected from the Bactrian camels and from environmental sources. Of these, 84 were morphologically identified as belonging to the Rhipicephalus sanguineus group and genetically identified (12S rDNA, 16S rDNA and the cytochrome c oxidase 1 genes) as R. sanguineus group ticks (temporally designated as Rhipicephalus sp. Xinjiang). PCR testing showed that 7.2% (20/279) of the camels harbored Anaplasma platys DNA. However, microscopic examination revealed no A. platys inclusions in blood smears from the camels. The PCR prevalence of A. platys DNA was 9.5% (6/63) in Rhipicephalus sp. Xinjiang from the Bactrian camels and 14.3% (3/21) in Rhipicephalus sp. Xinjiang from the vegetation. A. platys DNA was not detected by PCR in other tick species (Hyalomma asiaticum, Dermacentor niveus and Hyalomma dromedarii), and no other Anaplasma species were detected in these samples. This is the first report of A. platys in Bactrian camels in Xinjiang, China. The moderate positivity observed indicates that these animals might be a natural host for this pathogen in China.

Production and characterization of anti-(mucin MUC1) single-domain antibody in tobacco (Nicotiana tabacum cultivar Xanthi).

PubMed

Ismaili, Ahmad; Jalali-Javaran, Mokhtar; Rasaee, Mohammad J; Rahbarizadeh, Fatemeh; Forouzandeh-Moghadam, Mehdi; Memari, Hamid Rajabi

2007-05-01

Members of the Camelidae (camels, dromedaries, llamas, alpacas, guanacos and vicunas) are known to produce Igs (immunoglobulins) devoid of light chains and CH1s (constant heavy-chain domains). The antigen-specific binding fragments of these heavy-chain antibodies therefore comprise one single domain (the so-called 'VHH') and are of great importance in biotechnological applications. To evaluate the expression and biological activity of sdAbs (single-domain antibodies) in plants, which, on account of their small size and antigen-recognition properties, would have a major impact on antibody-engineering strategies, we constructed a pBI121-VHH gene encoding the recombinant sdAb fragments with specificity for a cancer-associated mucin, MUC1. Analysis of transgenic tobacco (Nicotiana tabacum cultivar Xanthi) plants by PCR and Western blotting demonstrated the expression of sdAb, while ELISA results with various MUC1 antigens and immunocytochemistry with cancerous cell lines confirmed that the activity of these molecules compared favourably with that of the parent recombinant antibodies. Protein purification was achieved by using sequential (NH4)2SO4 precipitation, gel filtration and immunoaffinity chromatography. Analysis of the purified VHH by ELISA indicated that the purified antibody fragments were able to react successfully with a MUC1-related peptide. These results reaffirm that the tobacco plant is a suitable host for the production of correctly folded VHH antibody fragments with diagnostic and therapeutic applications.

Genomic Data from Extinct North American Camelops Revise Camel Evolutionary History.

PubMed

Heintzman, Peter D; Zazula, Grant D; Cahill, James A; Reyes, Alberto V; MacPhee, Ross D E; Shapiro, Beth

2015-09-01

Recent advances in paleogenomic technologies have enabled an increasingly detailed understanding of the evolutionary relationships of now-extinct mammalian taxa. However, a number of enigmatic Quaternary species have never been characterized with molecular data, often because available fossils are rare or are found in environments that are not optimal for DNA preservation. Here, we analyze paleogenomic data extracted from bones attributed to the late Pleistocene western camel, Camelops cf. hesternus, a species that was distributed across central and western North America until its extinction approximately 13,000 years ago. Despite a modal sequence length of only around 35 base pairs, we reconstructed high-coverage complete mitochondrial genomes and low-coverage partial nuclear genomes for each specimen. We find that Camelops is sister to African and Asian bactrian and dromedary camels, to the exclusion of South American camelids (llamas, guanacos, alpacas, and vicuñas). These results contradict previous morphology-based phylogenetic models for Camelops, which suggest instead a closer relationship between Camelops and the South American camelids. The molecular data imply a Late Miocene divergence of the Camelops clade from lineages that separately gave rise to the extant camels of Eurasia. Our results demonstrate the increasing capacity of modern paleogenomic methods to resolve evolutionary relationships among distantly related lineages. © The Author 2015. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

1995 epizootic of vesicular stomatitis (New Jersey serotype) in the western United States: an entomologic perspective.

PubMed

Schmidtmann, E T; Tabachnick, W J; Hunt, G J; Thompson, L H; Hurd, H S

1999-01-01

Entomologic and epizootic data are reviewed concerning the potential for transmission of vesicular stomatitis (VS) virus by insects, including field data from case-positive premises in New Mexico and Colorado during the 1995 outbreak of the New Jersey serotype (VSNJ). As with previous outbreaks of VSNJ in the western United States, the 1995 epizootic illustrated that risk of exposure is seasonal, increasing during warm weather and decreasing with onset of cool weather; virus activity spread from south to north along river valleys of the southwestern and Rocky Mountain states; clinical disease was detected most commonly in horses, but also occurred in cattle and 1 llama; and most infections were subclinical. Overall, 367 case-positive premises were identified during the 1995 outbreak, with foci of virus activity along the Rio Grande River south of Albuquerque, NM, in southwestern Colorado, and along the Colorado River near Grand Junction, CO. The establishment of a 16-km (10-mile) radius zone of restricted animal movement around confirmed positive premises, along with imposition of state and international embargoes, created economic hardship for livestock owners and producers. The importance of defining the role of blood-feeding insects as biological vectors of VSNJ virus relative to risk factors that promote high levels of insect transmission, such as the presence of livestock along western river valleys, blood feeding activity, and frequent transport of animals for recreational purposes, is emphasized as a basis for developing effective disease management.

Trichophytobezoar duodenal obstruction in New World camelids.

PubMed

Sullivan, Eileen K; Callan, Robert J; Holt, Timothy N; Van Metre, David C

2005-01-01

To describe clinical findings, surgical treatment, and outcome associated with trichophytobezoar duodenal obstruction in New World camelids. Retrospective study. Alpacas (7) and 1 llama. Historical and clinical data were obtained from the medical records of New World camelids with a diagnosis of trichophytobezoar duodenal obstruction confirmed by surgical exploration or necropsy. Seven camelids were <1 year old. Abnormal clinical findings included anorexia, reduced fecal output, recumbency, colic, abdominal distension, regurgitation, decreased serum chloride concentration, increased serum bicarbonate concentration, and/or elevated first gastric compartment chloride concentration. Survey abdominal radiographs obtained (4 animals) revealed gastric distension (4) and/or visualization of the obstruction (2). Diagnosis was confirmed at necropsy (1) or surgery (7). Right paracostal celiotomy was performed on all animals and duodenotomy (3) or retropulsion of the trichophytobezoar combined with third compartment gastrotomy (4) was used to remove the obstruction. Six animals survived to discharge and 5 were healthy at follow-up, 8-20 months later. The remaining discharged alpaca was healthy at 12 months but subsequently died of unrelated causes. Diagnosis of trichophytobezoar duodenal obstruction should be considered in juvenile New World camelids with abdominal distension and hypochloremic metabolic alkalosis. Right paracostal celiotomy can be used for access to the descending duodenum and third gastric compartment for surgical relief of obstruction. Duodenal obstruction from bezoars should be considered in New World camelids <1year of age with abdominal distension and hypochloremic metabolic alkalosis. Surgical relief of the obstruction by right paracostal celiotomy has a good prognosis.

Development of VHH antibodies against dengue virus type 2 NS1 and comparison with monoclonal antibodies for use in immunological diagnosis.

PubMed

Fatima, Aneela; Wang, Haiying; Kang, Keren; Xia, Liliang; Wang, Ying; Ye, Wei; Wang, Jufang; Wang, Xiaoning

2014-01-01

The possibility of using variable domain heavy-chain antibodies (VHH antibodies) as diagnostic tools for dengue virus (DENV) type 2 NS1 protein was investigated and compared with the use of conventional monoclonal antibodies. After successful expression of DENV type 2 NS1 protein, the genes of VHH antibodies against NS1 protein were biopanned from a non-immune llama library by phage display. VHH antibodies were then expressed and purified from Escherichia coli. Simultaneously, monoclonal antibodies were obtained by the conventional route. Sequence analysis of the VHH antibodies revealed novel and long complementarity determining regions 3 (CDR3). Epitope mapping was performed via a phage display peptide library using purified VHH and monoclonal antibodies as targets. Interestingly, the same region of NS1, which comprises amino acids 224HWPKPHTLW232, was conserved for both kinds of antibodies displaying the consensus motif histidine-tryptophan-tryptophan or tryptophan-proline-tryptophan. The two types of antibodies were used to prepare rapid diagnostic kits based on immunochromatographic assay. The VHH antibody immobilized rapid diagnostic kit showed better sensitivity and specificity than the monoclonal antibody immobilized rapid diagnostic kit, which might be due to the long CDR3 regions of the VHH antibodies and their ability to bind to the pocket and cleft of the targeted antigen. This demonstrates that VHH antibodies are likely to be an option for developing point-of-care tests against DENV infection.

Development of VHH Antibodies against Dengue Virus Type 2 NS1 and Comparison with Monoclonal Antibodies for Use in Immunological Diagnosis

PubMed Central

Fatima, Aneela; Wang, Haiying; Kang, Keren; Xia, Liliang; Wang, Ying; Ye, Wei; Wang, Jufang; Wang, Xiaoning

2014-01-01

The possibility of using variable domain heavy-chain antibodies (VHH antibodies) as diagnostic tools for dengue virus (DENV) type 2 NS1 protein was investigated and compared with the use of conventional monoclonal antibodies. After successful expression of DENV type 2 NS1 protein, the genes of VHH antibodies against NS1 protein were biopanned from a non-immune llama library by phage display. VHH antibodies were then expressed and purified from Escherichia coli. Simultaneously, monoclonal antibodies were obtained by the conventional route. Sequence analysis of the VHH antibodies revealed novel and long complementarity determining regions 3 (CDR3). Epitope mapping was performed via a phage display peptide library using purified VHH and monoclonal antibodies as targets. Interestingly, the same region of NS1, which comprises amino acids 224HWPKPHTLW232, was conserved for both kinds of antibodies displaying the consensus motif histidine-tryptophan-tryptophan or tryptophan-proline-tryptophan. The two types of antibodies were used to prepare rapid diagnostic kits based on immunochromatographic assay. The VHH antibody immobilized rapid diagnostic kit showed better sensitivity and specificity than the monoclonal antibody immobilized rapid diagnostic kit, which might be due to the long CDR3 regions of the VHH antibodies and their ability to bind to the pocket and cleft of the targeted antigen. This demonstrates that VHH antibodies are likely to be an option for developing point-of-care tests against DENV infection. PMID:24751715

Variability in precipitation in a watershed in the altiplano, Peru and modes of variation

NASA Astrophysics Data System (ADS)

Mazzarino, M.; Brown, C. M.

2012-12-01

This research examines system linkages between climate, water availability, pasture availability, camelids (llamas and alpacas) and indigenous herders in an Andean watershed in southern Peru. In this region, extreme meteorological events such as drought and flood, occur often and have the potential to negatively impact herding livelihoods. Predictability in the system is paramount to reducing risks associated with these events. In the altiplano, a large portion of variability in precipitation has been attributed to the influence of El Nino Southern Oscillation (ENSO). In light of climate change and observations by herders, this research returns to the question of teleconnections in the altiplano. We use December through March precipitation totals obtained from eight meteorological stations for 43 years (1964-2006) and sea surface temperatures (SSTs) in the equatorial Pacific and Atlantic to characterize the hydroclimatology in the watershed and determine modes of variability. Following principal components analysis, prevailing periodicities in regional precipitation were determined using wavelet analysis and spatial correlation and regression analysis were used to determine the relationship between SST anomalies (SSTA's) and precipitation events in the watershed. Results suggest a non-linear and non-stationary mode of variability. We draw three conclusions from the results: 1) Positive precipitation extremes are dominated by an ENSO signal in the Nino 2 region; 2) Post 1987 there is a weak relationship, if any, between anomalously dry years in the precipitation record and SSTA's in the equatorial Pacific; 3) There is a stronger relationship (inverse) between precipitation in the region and SSTA's in the tropical Atlantic than previously believed.

Successful pregnancies from vitrified embryos in the dromedary camel: Avoidance of a possible toxic effect of sucrose on embryos.

PubMed

Herrid, M; Billah, M; Skidmore, J A

2017-12-01

Successful embryo cryopreservation facilitates the wider application of assisted reproduction technologies and also provides a useful method for gene banking of valuable genetics. Unfortunately attempts to establish an effective cryopreservation protocol for camelid embryos have been unsuccessful. In the current study, a modified vitrification protocol with three steps was investigated, whereby embryos were exposed to solutions containing increasing amounts of glycerol and ethylene glycol for fixed time periods. Embryos were then loaded into an Open Pull Straw (OPS) and plunged directly into liquid nitrogen for storage. Three experiments were designed to investigate the effect of 1) artificial shrinkage (AS) of embryos, 2) the addition of sucrose to the vitrification solutions, and 3) the replacement of sucrose by galactose in the warming solution, on the outcome of vitrification. The results showed that neither AS of hatched embryos prior to vitrification, nor the addition of sucrose into vitrification solutions improves the outcome of vitrification, while replacement of sucrose with galactose in warming solution increases the survival and developmental rates of vitrified embryos in culture. Transfer of vitrified embryos that were warmed in galactose resulted in a pregnancy rate of 42.8% per embryo or 46.1% per recipient. Collectively, these results suggest a possible species-specific toxic effect of sucrose on camel embryos, and that avoiding its use either in vitrification or warming solution is critical for establishing an effective protocol. This study may also be applicable to the vitrification of embryos of other camelid species including alpaca and llamas. Copyright © 2017 Elsevier B.V. All rights reserved.

LLAMA: nuclear stellar properties of Swift-BAT AGN and matched inactive galaxies

NASA Astrophysics Data System (ADS)

Lin, Ming-Yi; Davies, R. I.; Hicks, E. K. S.; Burtscher, L.; Contursi, A.; Genzel, R.; Koss, M.; Lutz, D.; Maciejewski, W.; Müller-Sánchez, F.; Orban de Xivry, G.; Ricci, C.; Riffel, R.; Riffel, R. A.; Rosario, D.; Schartmann, M.; Schnorr-Müller, A.; Shimizu, T.; Sternberg, A.; Sturm, E.; Storchi-Bergmann, T.; Tacconi, L.; Veilleux, S.

2018-02-01

In a complete sample of local 14-195 keV selected active galactic nuclei (AGNs) and inactive galaxies, matched by their host galaxy properties, we study the spatially resolved stellar kinematics and luminosity distributions at near-infrared wavelengths on scales of 10-150 pc, using SINFONI on the VLT. In this paper, we present the first half of the sample, which comprises 13 galaxies, eight AGNs and five inactive galaxies. The stellar velocity fields show a disc-like rotating pattern, for which the kinematic position angle is in agreement with the photometric position angle obtained from large scale images. For this set of galaxies, the stellar surface brightness of the inactive galaxy sample is generally comparable to the matched sample of AGN, but extends to lower surface brightness. After removal of the bulge contribution, we find a nuclear stellar light excess with an extended nuclear disc structure, which exhibits a size-luminosity relation. While we expect the excess luminosity to be associated with a dynamically cooler young stellar population, we do not typically see a matching drop in dispersion. This may be because these galaxies have pseudo-bulges in which the intrinsic dispersion increases towards the centre. And although the young stars may have an impact in the observed kinematics, their fraction is too small to dominate over the bulge and compensate the increase in dispersion at small radii, so no dispersion drop is seen. Finally, we find no evidence for a difference in the stellar kinematics and nuclear stellar luminosity excess between these active and inactive galaxies.

Acute gastrointestinal disease in 27 New World camelids: clinical and surgical findings.

PubMed

Cebra, C K; Cebra, M L; Garry, F B; Larsen, R S; Baxter, G M

1998-01-01

To describe clinical and surgical findings from New World camelids with acute gastrointestinal disease. Retrospective study. 20 llamas and 7 alpacas. Camelids were grouped based on surgical lesions. Clinical and surgical findings were compared between groups and between surviving and nonsurviving camelids. Twelve of 27 initial celiotomies and 3 of 4 repeat celiotomies were successful. Death occurred from euthanasia during surgery (nine camelids), peritonitis or sepsis (five), aspiration pneumonia (one), and respiratory distress (one). Survival was lowest after celiotomy for proximal obstruction (3 of 10 camelids), ruptured viscus (0 of 4), and necrotizing enteritis (0 of 2) and highest after celiotomy for distal obstruction (10 of 13) and septic peritonitis without ruptured viscus (2 of 2). Before surgery, camelids with proximal obstruction had significantly lower (P < .05) serum chloride concentrations (median, 97 mEq/L) than those with distal obstruction (median, 109 mEq/L) or ruptured viscus (median, 117 mEq/L). Serum bicarbonate concentration also was highest (median, 34.6 mEq/L) and often greater than 28 mEq/L in camelids with proximal obstruction. Camelids with distal obstruction had significantly lower (P < .05) nucleated cell counts in peritoneal fluid (median, 700 cells/microL) than those with ruptured viscus (median, 20,600 cells/microL) or septic peritonitis (median, 88,300 cells/microL). Camelids with proximal obstruction often had hypochloremic metabolic alkalosis. Camelids with distal obstruction had less metabolic derangement and tissue compromise and a higher survival rate. Awareness of the characteristics of the various types of acute gastrointestinal disease in camelids will augment veterinarians' ability to diagnose and treat these disorders.

Computed tomographic and radiographic examination of dental structures in South American camelid specimen of different ages.

PubMed

Rostami, Abdolrazagh; Geissbühler, Urs; Schellenberger, Frank; Zanolari, Patrik

2014-01-06

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 protocol, radiographs and CT scans of the 20 specimen were performed. The most useful radiographic projections for mandibular and maxillary cheek teeth evaluation turned out to be lateral30°ventral-laterodorsal and lateral30°dorsal-lateroventral with slight separation of the dental arcades respectively. Digital radiographic and CT appearance of the mandibular and maxillary teeth were described from the beginning of mineralization till maturity. In addition the normal range of the CT radio density of different cheek teeth and different dental tissues were measured. Hounsfield units of different dental tissues of SAC turned out to be similar to equids. Deviation, shortening and partial destruction of the distal tooth root of mandibular 09's and 10's and of maxillary 09's was observed and the existence of a common pulp chamber in younger teeth was revealed. The present study provides information about the dental imaging morphology in clinically healthy SAC. This basic information provides fundamental knowledge for evaluating images and planning treatments in clinically affected animals.

Development and Evaluation of Single Domain Antibodies for Vaccinia and the L1 Antigen

PubMed Central

Walper, Scott A.; Liu, Jinny L.; Zabetakis, Daniel; Anderson, George P.; Goldman, Ellen R.

2014-01-01

There is ongoing interest to develop high affinity, thermal stable recognition elements to replace conventional antibodies in biothreat detection assays. As part of this effort, single domain antibodies that target vaccinia virus were developed. Two llamas were immunized with killed viral particles followed by boosts with the recombinant membrane protein, L1, to stimulate the immune response for envelope and membrane proteins of the virus. The variable domains of the induced heavy chain antibodies were selected from M13 phage display libraries developed from isolated RNA. Selection via biopanning on the L1 antigen produced single domain antibodies that were specific and had affinities ranging from 4×10−9 M to 7.0×10−10 M, as determined by surface plasmon resonance. Several showed good ability to refold after heat denaturation. These L1-binding single domain antibodies, however, failed to recognize the killed vaccinia antigen. Useful vaccinia binding single domain antibodies were isolated by a second selection using the killed virus as the target. The virus binding single domain antibodies were incorporated in sandwich assays as both capture and tracer using the MAGPIX system yielding limits of detection down to 4×105 pfu/ml, a four-fold improvement over the limit obtained using conventional antibodies. This work demonstrates the development of anti-vaccinia single domain antibodies and their incorporation into sandwich assays for viral detection. It also highlights the properties of high affinity and thermal stability that are hallmarks of single domain antibodies. PMID:25211488

Evaluation of sensitivity and specificity of cytologic examination: 269 cases (1999-2000).

PubMed

Cohen, Michele; Bohling, Mark W; Wright, James C; Welles, Elizabeth A; Spano, Joseph S

2003-04-01

To determine sensitivity and specificity of cytologic examination used in a clinical setting. Retrospective study. 216 dogs, 44 cats, 4 horses, 2 ferrets, 1 llama, 1 rat, and 1 mouse. Records were reviewed of cases in which a cytologic diagnosis was followed by a surgical biopsy or postmortem examination within 3 days with subsequent histopathologic diagnosis. Diagnoses were compared for agreement at various levels, including complete agreement, partial agreement, no agreement, or no comparison possible because of insufficient or incorrect cytologic specimen. Levels of agreement were compared for different categories of lesions, including neoplastic, inflammatory, dysplastic-hyperplastic-other, and normal tissue. Additionally, levels of agreement for neoplastic lesions were categorized with regard to cell type, degree of malignancy, and location. Sensitivity and specificity of cytologic examination were calculated. At the level of general agreement (complete and partial agreement), the sensitivity of cytologic examination ranged from 33.3 to 66.1%, depending on the location of the lesion. Cytologic examination was most accurate when used to diagnose cutaneous and subcutaneous lesions and least accurate for diagnosis of liver lesions. Cytologic examination was most effective in diagnosis of neoplastic disease and least effective in diagnosis of dysplastic or hyperplastic conditions. Cytologic examination is a valuable diagnostic tool, although our results indicated lower accuracy than previously reported. False-negative results (missing a diagnosis) were far more common than false-positive results (categorizing a healthy animal as diseased): therefore, if the clinical index of suspicion is high, cytologic examination should be repeated or another technique should be selected to rule out the suspected condition.

Computed tomographic and radiographic examination of dental structures in South American camelid specimen of different ages

PubMed Central

2014-01-01

Background 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 protocol, radiographs and CT scans of the 20 specimen were performed. Results The most useful radiographic projections for mandibular and maxillary cheek teeth evaluation turned out to be lateral30°ventral - laterodorsal and lateral30°dorsal - lateroventral with slight separation of the dental arcades respectively. Digital radiographic and CT appearance of the mandibular and maxillary teeth were described from the beginning of mineralization till maturity. In addition the normal range of the CT radio density of different cheek teeth and different dental tissues were measured. Hounsfield units of different dental tissues of SAC turned out to be similar to equids. Deviation, shortening and partial destruction of the distal tooth root of mandibular 09′s and 10′s and of maxillary 09′s was observed and the existence of a common pulp chamber in younger teeth was revealed. Conclusions The present study provides information about the dental imaging morphology in clinically healthy SAC. This basic information provides fundamental knowledge for evaluating images and planning treatments in clinically affected animals. PMID:24393365

Physiological studies in the South American camelid llama (Lama guanicoe f. d. glama). I. Body water spaces and water turnover.

PubMed

Marcilese, N A; Ghezzi, M D; Aba, M A; Alzola, R A; Solana, H; Valsecchi, R M

1994-01-01

Body water (BW) and extracellular water (ECW), were determined with tritiated water (THO) and 82Br injected into the vein, to 8 mature animals of both sexes during the winter season. The biological half-time of THO (T1/2 THO) and the daily water turnover (WT) were measured and the intracellular water (ICW) calculated. The studies with THO were repeated in the same animals and in 2 lactating females in spring and summer. Two calves were also studied during spring. The values obtained in winter were: BW 659 +/- 12 ml/kg; T1/2 THO 9.2 +/- 1 day; WT per 24 h 50 +/- 3 ml/kg or 116 +/- 5 ml/kg 82 and 163 +/- 9 ml per 1 of BW82; ECW 215 +/- 8 ml/kg or 32.5 +/- 3% BW; ICW 447 +/- 12 ml/kg or 67.7 +/- 4% BW. The results of the spring's studies showed a significant increase in the values of WT. In summer a further increment of this parameters was observed when expressed as ml/kg body solids. This differences were remarkable in those in lactation. The proportion of water in the body was significantly higher during summer in all animals. BW in lactating animals during summer was 783 +/- 9 ml/kg and in the other animals 718 +/- 18; T1/2 THO values were 3.3 +/- .-06 and 4.5 +/- .4 day, respectively. WT was 396 +/- 9 ml/kg.82 or 484 +/- 8 ml/l BW82 in the lactating animals and 260 +/- 9 ml/kg 82 or 341 +/- 12 ml/l BW82 in the other animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Next-Generation DNA Sequencing of VH/VL Repertoires: A Primer and Guide to Applications in Single-Domain Antibody Discovery.

PubMed

Henry, Kevin A

2018-01-01

Immunogenetic analyses of expressed antibody repertoires are becoming increasingly common experimental investigations and are critical to furthering our understanding of autoimmunity, infectious disease, and cancer. Next-generation DNA sequencing (NGS) technologies have now made it possible to interrogate antibody repertoires to unprecedented depths, typically by sequencing of cDNAs encoding immunoglobulin variable domains. In this chapter, we describe simple, fast, and reliable methods for producing and sequencing multiplex PCR amplicons derived from the variable regions (V H , V H H or V L ) of rearranged immunoglobulin heavy and light chain genes using the Illumina MiSeq platform. We include complete protocols and primer sets for amplicon sequencing of V H /V H H/V L repertoires directly from human, mouse, and llama lymphocytes as well as from phage-displayed V H /V H H/V L libraries; these can be easily be adapted to other types of amplicons with little modification. The resulting amplicons are diverse and representative, even using as few as 10 3 input B cells, and their generation is relatively inexpensive, requiring no special equipment and only a limited set of primers. In the absence of heavy-light chain pairing, single-domain antibodies are uniquely amenable to NGS analyses. We present a number of applications of NGS technology useful in discovery of single-domain antibodies from phage display libraries, including: (i) assessment of library functionality; (ii) confirmation of desired library randomization; (iii) estimation of library diversity; and (iv) monitoring the progress of panning experiments. While the case studies presented here are of phage-displayed single-domain antibody libraries, the principles extend to other types of in vitro display libraries.

Speciation of arsenic in bulk and rhizosphere soils from artisanal cooperative mines in Bolivia.

PubMed

Acosta, Jose A; Arocena, Joselito M; Faz, Angel

2015-11-01

Soils near artisanal and small-scale gold mines (ASGM) have high arsenic (As) contents due to the presence of arsenopyrite in gold ores and accelerated accumulations due to mine wastes disposal practices and other mining activities. We determined the content and speciation to understand the fate and environmental risks of As accumulations in 24 bulk and 12 rhizosphere soil samples collected in the Virgen Del Rosario and the Rayo Rojo cooperative mines in the highlands of Bolivia. Mean total As contents in bulk and rhizosphere soils ranged from 13 to 64 mg kg(-1) and exceeded the soil environmental quality guidelines of Canada. Rhizosphere soils always contained at least twice the As contents in the bulk soil. Elemental mapping using 4×5 μm synchrotron-generated X-ray micro-beam revealed As accumulations in areas enriched with Fe. Results of As-X-ray Absorption Near Edge Spectroscopy (As-XANES) showed that only As(V) species was detectable in all samples regardless of As contents, size fractions and types of vegetation. Although the toxicity of As(V) is less than As(III), we suggest that As uptake of commonly-grazed vegetation by alpaca and llama must be determined to fully understand the environmental risks of high As in soils near ASGM in Bolivia. In addition, knowledge on the speciation of the As bio-accessible fraction will provide another useful information to better understand the fate and transfer of As from soils into the food chain in environments associated with the ASGM in Bolivia and other parts of the world. Copyright © 2014 Elsevier Ltd. All rights reserved.

Therapeutics and Immunoprophylaxis Against Noroviruses and Rotaviruses: The Past, Present, and Future

PubMed Central

Ghosh, Souvik; Malik, Yashpal Singh; Kobayashi, Nobumichi

2018-01-01

Background: Noroviruses and rotaviruses are important viral etiologies of severe gastroenteritis. Noroviruses are the primary cause of nonbacterial diarrheal outbreaks in humans, whilst rotaviruses are a major cause of childhood diarrhea. Although both enteric pathogens substantially impact human health and economies, there are no approved drugs against noroviruses and rotaviruses so far. On the other hand, whilst the currently licensed rotavirus vaccines have been successfully implemented in over 100 countries, the most advanced norovirus vaccine has recently completed phase-I and II trials. Methods: We performed a structured search of bibliographic databases for peer-reviewed research litera-ture on advances in the fields of norovirus and rotavirus therapeutics and immunoprophylaxis. Results: Technological advances coupled with a proper understanding of viral morphology and replication over the past decade has facilitated pioneering research on therapeutics and immunoprophylaxis against noroviruses and rotaviruses, with promising outcomes in human clinical trials of some of the drugs and vaccines. This review focuses on the various developments in the fields of norovirus and rotavirus thera-peutics and immunoprophylaxis, such as potential antiviral drug molecules, passive immunotherapies (oral human immunoglobulins, egg yolk and bovine colostral antibodies, llama-derived nanobodies, and anti-bodies expressed in probiotics, plants, rice grains and insect larvae), immune system modulators, probiot-ics, phytochemicals and other biological substances such as bovine milk proteins, therapeutic nanoparti-cles, hydrogels and viscogens, conventional viral vaccines (live and inactivated whole virus vaccines), and genetically engineered viral vaccines (reassortant viral particles, virus-like particles (VLPs) and other sub-unit recombinant vaccines including multi-valent viral vaccines, edible plant vaccines, and encapsulated viral particles). Conclusions: This review provides important insights into the various approaches to therapeutics and im-munoprophylaxis against noroviruses and rotaviruses.. PMID:28901254

Hybridizing Old and New World camelids: Camelus dromedarius x Lama guanicoe.

PubMed Central

Skidmore, J A; Billah, M; Binns, M; Short, R V; Allen, W R

1999-01-01

Thirty female dromedary camels were inseminated on a total of 50 occasions with 2-4 ml of fresh guanaco semen diluted with an equal volume of commercially available camel semen extender. Similarly, nine female guanacos were inseminated on 34 occasions with 4-6 ml of fresh, diluted camel semen. Only two of the dromedary females conceived; one aborted a female foetus on day 260 of gestation and the other gave birth to a stillborn female calf on day 365. Six conceptions occurred in the female guanacos. Two of these conceptuses, diagnosed by ultrasound, were resorbed between days 25 and 40 of gestation, one female foetus was aborted on day 291, another female foetus was aborted on day 302, and one female calf was stillborn on day 365 of gestation. The sixth foetus, a male, was born prematurely but alive after a 328-day gestation. It had a phenotypic appearance intermediate between that of a camel and a guanaco and its hybrid parentage was confirmed by the DNA fingerprinting of eight llama microsatellites. To our knowledge, this is the first viable hybrid ever to be produced between Old World and New World camelids, which have been reproductively isolated from one another for at least 11 million years. The preponderance of female hybrids is in accordance with Haldane's law. Histological examination of their ovaries revealed a failure of meiosis, with only an occasional abnormal oocyte surrounded by follicle cells. Although the diploid chromosone number of camels and guanacos is the same (2n = 74), sufficient genetic change has taken place to make the pairing of homologous chromosomes no longer possible. PMID:10331286

β-Nerve growth factor is a major component of alpaca seminal plasma and induces ovulation in female alpacas.

PubMed

Kershaw-Young, C M; Druart, X; Vaughan, J; Maxwell, W M C

2012-01-01

Ovulation in camelids is induced by an unidentified protein in the seminal plasma of the male termed 'ovulation-inducing factor'. This protein has been reported to be a 14-kDa protein under reducing conditions, which, when purified from seminal plasma, induces ovulation in llamas. The identification of this protein and investigation of its potential to induce ovulation in camelids may aid the development of protocols for the induction of ovulation. In the present study, alpaca seminal plasma proteins were separated using one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the most abundant protein of 14 kDa was identified as β-nerve growth factor (β-NGF) by liquid chromatography mass spectrometry. Female alpacas (n = 5 per group) were given intramuscular injections of: (1) 1 mL of 0.9% saline; (2) 4 µg buserelin, a gonadotrophin-releasing hormone agonist; (3) 2 mL alpaca seminal plasma; or (4) 1mg human β-NGF. Ovulation was detected by transrectal ultrasonography 8 days after treatment and confirmed by plasma progesterone concentrations. Ovulation occurred in 0%, 80%, 80% and 80% of animals treated with saline, buserelin, seminal plasma and β-NGF, respectively. Treatment type did not affect the diameter of the corpus luteum, but plasma progesterone concentrations were lower in saline-treated animals than in the other treatment groups owing to the lack of a corpus luteum. The present study is the first to identify the ovulation-inducing factor protein in alpacas. β-NGF successfully induces ovulation in alpacas and this finding may lead to new methods for the induction of ovulation in camelids.

Schmallenberg virus infection in South American camelids: Field and experimental investigations.

PubMed

Schulz, Claudia; Beer, Martin; Hoffmann, Bernd

2015-11-18

During the first epizootic wave of the novel, teratogenic Schmallenberg virus (SBV, Orthobunyavirus) in ruminants in Northern Europe, serological evidence of a previous SBV-infection demonstrated that South American camelids (SAC) are also susceptible to SBV. However, their potential role in SBV spread remains unknown. To investigate the prevalence and course of SBV-infection in SAC, a German field study and an animal trial with three llamas and three alpacas were conducted. From September 2012 to December 2013, 313 of 502 SAC (62.35%) were found SBV seropositive, but negative for SBV-RNA. The estimated between-district (94.23% of 52) and median within-district (71.43%) and herd (73.13%) SBV seroprevalence in German SAC was similar to the seroprevalence reported in cattle herds and sheep flocks at the time. An age of >1 year was found a statistically significant risk factor for SBV-infection, which could be explained by the spatio-temporal spread of SBV in Germany during the study period. No clinical signs or an increase of abortion and congenital malformation associated with SBV-infection in SAC were reported by the study participants. Similar to SBV-infected ruminants, SBV-RNAemia in experimentally SBV-infected SAC was detected for a short time between days 3 and 7 after infection (dpi), and seroconversion occurred between 9 and 21 dpi. Despite the similar virological and serological results, the lack of clinical signs and congenital malformation associated with SBV-infection suggests that SBV causes subclinical infection in SAC. However, their role as reservoirs in the spread of SBV has to be further investigated. Copyright © 2015 Elsevier B.V. All rights reserved.

Camelid Ig V genes reveal significant human homology not seen in therapeutic target genes, providing for a powerful therapeutic antibody platform

PubMed Central

Klarenbeek, Alex; Mazouari, Khalil El; Desmyter, Aline; Blanchetot, Christophe; Hultberg, Anna; de Jonge, Natalie; Roovers, Rob C; Cambillau, Christian; Spinelli, Sylvia; Del-Favero, Jurgen; Verrips, Theo; de Haard, Hans J; Achour, Ikbel

2015-01-01

Camelid immunoglobulin variable (IGV) regions were found homologous to their human counterparts; however, the germline V repertoires of camelid heavy and light chains are still incomplete and their therapeutic potential is only beginning to be appreciated. We therefore leveraged the publicly available HTG and WGS databases of Lama pacos and Camelus ferus to retrieve the germline repertoire of V genes using human IGV genes as reference. In addition, we amplified IGKV and IGLV genes to uncover the V germline repertoire of Lama glama and sequenced BAC clones covering part of the Lama pacos IGK and IGL loci. Our in silico analysis showed that camelid counterparts of all human IGKV and IGLV families and most IGHV families could be identified, based on canonical structure and sequence homology. Interestingly, this sequence homology seemed largely restricted to the Ig V genes and was far less apparent in other genes: 6 therapeutically relevant target genes differed significantly from their human orthologs. This contributed to efficient immunization of llamas with the human proteins CD70, MET, interleukin (IL)-1β and IL-6, resulting in large panels of functional antibodies. The in silico predicted human-homologous canonical folds of camelid-derived antibodies were confirmed by X-ray crystallography solving the structure of 2 selected camelid anti-CD70 and anti-MET antibodies. These antibodies showed identical fold combinations as found in the corresponding human germline V families, yielding binding site structures closely similar to those occurring in human antibodies. In conclusion, our results indicate that active immunization of camelids can be a powerful therapeutic antibody platform. PMID:26018625

Streptococcus equi subsp zooepidemicus pleuropneumonia and peritonitis in a dromedary camel (Camelus dromedarius) calf in North America.

PubMed

Stoughton, William B; Gold, Jenifer

2015-08-01

A 12-week-old female dromedary camel (Camelus dromedarius) calf was evaluated because of acute (< 24 hours) inappetence and lethargy. The calf was being bottle-fed because of maternal rejection. Physical examination revealed decreased bronchovesicular sounds and absent borborygmi. The rectal temperature was 38.9°C (102.0°F). A CBC indicated leukopenia with a degenerative left shift suggestive of a systemic infection. Results of abdominal and thoracic ultrasonography showed severe bicavitary effusion, peripheral lung consolidation, and intestinal hypomotility. Pleural and peritoneal fluid analysis confirmed a diagnosis of septic pleuritis and peritonitis. Results of aerobic bacterial culture of venous blood, peritoneal fluid, and pleural fluid samples indicated Streptococcus equi subsp zooepidemicus septicemia as the etiology for the polyserositis (ie, alpaca fever). Treatment with IV broad-spectrum antimicrobials, an NSAID, and pleural drainage was initiated. Clinical signs of pleuropneumonia, peritonitis, and systemic infection improved rapidly 24 hours after initiation of medical treatment. The calf was discharged from the hospital after 11 days, and antimicrobial treatment continued for 2 weeks after discharge. At follow-up approximately 4 weeks after hospital discharge (6 weeks after the initial examination), there were no clinical signs suggestive of relapse or any reported complications. S equi subsp zooepidemicus may cause polyserositis in Old World camelids (eg, dromedary camels) with signs similar to those seen in New World camelids (eg, alpaca and llama). The rapid response to medical treatment for the patient described suggested that S equi subsp zooepidemicus-induced polyserositis (alpaca fever) in dromedary camels may respond favorably to appropriate treatment. Reducing stress, reducing overcrowding, and separate housing of equids and camelids are suggested. Further studies are needed to better assess the epidemiology of alpaca fever in dromedary camels in North America.

Curved bones: An adaptation to habitual loading.

PubMed

Milne, Nick

2016-10-21

Why are long bones curved? It has long been considered a paradox that many long bones supporting mammalian bodies are curved, since this curvature results in the bone undergoing greater bending, with higher strains and so greater fracture risk under load. This study develops a theoretical model wherein the curvature is a response to bending strains imposed by the requirements of locomotion. In particular the radioulna of obligate quadrupeds is a lever operated by the triceps muscle, and the bending strains induced by the triceps muscle counter the bending resulting from longitudinal loads acting on the curved bone. Indeed the theoretical model reverses this logic and suggests that the curvature is itself a response to the predictable bending strains induced by the triceps muscle. This, in turn, results in anatomical arrangements of bone, muscle and tendon that create a simple physiological mechanism whereby the bone can resist the bending due to the action of triceps in supporting and moving the body. The model is illustrated by contrasting the behaviour of a finite element model of a llama radioulna to that of a straightened version of the same bone. The results show that longitudinal and flexor muscle forces produce bending strains that effectively counter strains due to the pull of the triceps muscle in the curved but not in the straightened model. It is concluded that the curvature of these and other curved bones adds resilience to the skeleton by acting as pre-stressed beams or strainable pre-buckled struts. It is also proposed that the cranial bending strains that result from triceps, acting on the lever that is the radioulna, can explain the development of the curvature of such bones. Crown Copyright © 2016. Published by Elsevier Ltd. All rights reserved.

In vivo imaging in the rabbit as a model for the study of ovulation-inducing factors.

PubMed

Cervantes, M P; Palomino, J M; Adams, G P

2015-01-01

The study of factors responsible for eliciting ovulation in rabbits has been hampered by the lack of a suitable method of monitoring the ovaries in vivo. Ovarian imaging by ultrasound biomicroscopy was used in two experiments designed to determine the effects of seminal plasma on the ovulatory response in rabbits. In Experiment 1, female rabbits were group-housed and treated intramuscularly with saline, gonadotropin releasing hormone (GnRH), or seminal plasma of llamas or rabbits (n = 4 to 6 per group). Rabbits were euthanized eight days later to evaluate the ovarian response by ultrasound biomicroscopy ex situ. No differences among groups were detected in the proportion of rabbits that ovulated or in the number and size of corpora lutea. The high incidence of ovulation in the negative control group was unexpected, and confounded determination of an ovulation-inducing effect of seminal plasma. In Experiment 2, female rabbits were caged individually, and treated as in Experiment 1 (n = 5 to 7 per group). The ovarian response was evaluated in vivo by transcutaneous ultrasound biomicroscopy. Ovulation and formation of corpora lutea were detected only in rabbits given GnRH. A preovulatory surge in plasma luteinizing hormone concentration and a post-ovulatory rise in plasma progesterone concentration were detected only in rabbits treated with GnRH. Surgical translocation of the ovaries to a subcutaneous position enabled longitudinal assessment of the ovulatory response by ultrasound biomicroscopy. Results clearly documented the effect of physical/social interaction on ovulation in rabbits, and did not support the hypothesis that seminal plasma elicits ovulation in rabbits. © The Author(s) 2014 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

Inhibition of glycosylation on a camelid antibody uniquely affects its FcγRI binding activity.

PubMed

Krahn, Natalie; Spearman, Maureen; Meier, Markus; Dorion-Thibaudeau, July; McDougall, Matthew; Patel, Trushar R; De Crescenzo, Gregory; Durocher, Yves; Stetefeld, Jörg; Butler, Michael

2017-01-01

Glycoengineering of mAbs has become common practice in attempts to generate the ideal mAb candidate for a wide range of therapeutic applications. The effects of these glycan modifications on the binding affinity of IgG mAbs for FcγRIIIa and their cytotoxicity are well known. However, little is understood about the effect that these modifications have on binding to the high affinity FcγRI receptor. This study analyzed the effect of variable N-glycosylation on a human-llama hybrid mAb (EG2-hFc, 80kDa) binding to FcγRI including a comparison to a full-sized IgG1 (DP-12, 150kDa). This was achieved by the addition of three glycosylation inhibitors (swainsonine, castanospermine, and kifunensine) independently to Chinese hamster ovary (CHO) cell cultures to generate hybrid and high mannose glycan structures. Biophysical analysis by circular dichroism, dynamic light scattering and analytical ultra-centrifugation confirmed that the solution-behaviour of the mAbs remained constant over multiple concentrations and glycan treatments. However, changes were observed when studying the interaction of FcγRI with variously glycosylated mAbs. Both mAbs were observed to have a decreased binding affinity upon treatment with swainsonine which produced hybrid glycans. Following de-glycosylation the binding affinity for EG2-hFc was only marginally reduced (6-fold) compared to a drastic (118-fold) decrease for DP-12. In summary, our data suggest that the relatively low molecular weight of chimeric EG2-hFc may contribute to its enhanced stability against glycan changes making it a highly suitable mAb candidate for therapeutic applications. Copyright © 2016 Elsevier B.V. All rights reserved.

Low habitat overlap at landscape scale between wild camelids and feral donkeys in the Chilean desert

NASA Astrophysics Data System (ADS)

Malo, Juan E.; González, Benito A.; Mata, Cristina; Vielma, André; Donoso, Denise S.; Fuentes, Nicolás; Estades, Cristián F.

2016-01-01

Feral domestic ungulates may compete with the populations of wild herbivores with which they coexist, particularly so in arid regions. The potential competition between wild camelids and feral donkeys at the eastern sector of the Atacama Desert is evaluated in terms of their coincidence or segregation in habitat use and complemented with a comparison of reproductive output (yearling/adult ratio) of vicuña family groups in the proximity vs. distant from donkey observations. Habitat use of wild camelids and donkeys was sampled driving some 1250 km of roads and tracks at the dry and wet seasons. There were 221 vicuñas (Vicugna vicugna) sightings, 77 for donkeys (Equus asinus), 25 for guanacos (Lama guanicoe) and 8 for hybrids between guanacos and domestic llamas (Lama glama), as well as 174 randomly selected control locations. By means of Generalised Discriminant Analysis and Analysis of Variance we show that all ungulates actively select their habitat, with significant differences between use and availability in the area. Donkeys are relatively abundant in comparison with camelids and coincide broadly with both of them across the altitudinal gradient, but they fall between them in local scale habitat selection and do not seem to force their displacement from their preferred habitats. Thus donkeys occur preferentially on slopes with a high cover of tall shrubs, whereas vicuñas use valley bottoms with grass and guanacos the upper slope zones with grass. The potential for competition between donkeys and wild camelids is thus limited and it does not affect the reproductive output of vicuña in this region. Therefore, with the present knowledge we suggest that population control is not currently merited for feral donkeys.

Risk factors associated with anthrax outbreak in animals in North Dakota, 2005: a retrospective case-control study.

PubMed

Mongoh, Mafany Ndiva; Dyer, Neil W; Stoltenow, Charles L; Khaitsa, Margaret L

2008-01-01

We identified the risk factors associated with the anthrax outbreak Of 2005 in animals in North Dakota. Medical records of the 2005 anthrax outbreak were obtained from the Veterinary Diagnostic Laboratory at North Dakota State University. Additional data were obtained from the North Dakota state veterinarian's office, and supplemental questionnaires were administered to producers. The data obtained included ecological and environmental factors, animal health factors, and management factors. Anthrax occurred from July 1 to October 12, 2005. The cases were located in eastern North Dakota around the Red River Basin. Ransom, LaMoure, and Barnes counties reported most cases (71%). Species affected included cattle, bison, horses, sheep, elk, deer, pigs, and llamas. The predominant symptom was sudden death (38%) followed by bleeding from orifices (17%). Chi-square analysis indicated significant differences between case and control premises on the following variables: death reported on neighboring pasture, vaccination period, dry conditions, wet conditions, antibiotic use, multiple vaccination, and type of predator (coyote). Factors that significantly (p<0.05) predicted anthrax occurrences on the final logistic regression model were vaccination, use of antibiotics during an outbreak, and period of vaccine administration (before or during the outbreak). The characteristics of the anthrax outbreak regarding time and place of occurrence, animals affected, clinical signs reported, and mortality rate were consistent with previous reports of natural anthrax outbreaks in animals. A number of factors that significantly predicted anthrax occurrence in animals in the 2005 outbreak in North Dakota were identified. This information is important in planning appropriate control and prevention measures for anthrax, including recommending the right vaccination and treatment regimens in managing future anthrax outbreaks.

Therapeutics and Immunoprophylaxis Against Noroviruses and Rotaviruses: The Past, Present, and Future.

PubMed

Ghosh, Souvik; Malik, Yashpal Singh; Kobayashi, Nobumichi

2018-01-01

Noroviruses and rotaviruses are important viral etiologies of severe gastroenteritis. Noroviruses are the primary cause of nonbacterial diarrheal outbreaks in humans, whilst rotaviruses are a major cause of childhood diarrhea. Although both enteric pathogens substantially impact human health and economies, there are no approved drugs against noroviruses and rotaviruses so far. On the other hand, whilst the currently licensed rotavirus vaccines have been successfully implemented in over 100 countries, the most advanced norovirus vaccine has recently completed phase-I and II trials. We performed a structured search of bibliographic databases for peer-reviewed research literature on advances in the fields of norovirus and rotavirus therapeutics and immunoprophylaxis. Technological advances coupled with a proper understanding of viral morphology and replication over the past decade has facilitated pioneering research on therapeutics and immunoprophylaxis against noroviruses and rotaviruses, with promising outcomes in human clinical trials of some of the drugs and vaccines. This review focuses on the various developments in the fields of norovirus and rotavirus therapeutics and immunoprophylaxis, such as potential antiviral drug molecules, passive immunotherapies (oral human immunoglobulins, egg yolk and bovine colostral antibodies, llama-derived nanobodies, and antibodies expressed in probiotics, plants, rice grains and insect larvae), immune system modulators, probiotics, phytochemicals and other biological substances such as bovine milk proteins, therapeutic nanoparticles, hydrogels and viscogens, conventional viral vaccines (live and inactivated whole virus vaccines), and genetically engineered viral vaccines (reassortant viral particles, virus-like particles (VLPs) and other subunit recombinant vaccines including multi-valent viral vaccines, edible plant vaccines, and encapsulated viral particles). This review provides important insights into the various approaches to therapeutics and immunoprophylaxis against noroviruses and rotaviruses. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

Glacier Retreat in the Southern Peruvian Andes: Climate Change, Environmental Impacts, Human Perception and Social Response

NASA Astrophysics Data System (ADS)

Orlove, B.

2007-12-01

This paper presents results from recent environmental and anthropological research near glacierized areas in the department of Cusco, Peru, home to the well-known Quelccaya Ice Cap and to the peak of Ausangate (6384 m). Glaciers in the region are in negative mass balance, losing volume and area, with upslope movement of the glacier fronts. Somewhat paradoxically, flows in many streams close to the glaciers are reduced, particularly in the dry season, due to a shift in the seasonal distribution of melting, to increased evaporation and to increased percolation into newly-exposed sands and gravels. Associated with this reduction in flow is a desiccation of some anthropogenic and natural wetlands, reducing the availability of dry season forage to wild (vicuna) and domesticated (alpaca, llama) ruminants. Interviews and ethnographic observations with local populations of Quechua-speaking herders at elevations of 4500-5200 meters provide detailed comments on these changes. They have an extensive vocabulary of terms for glacial features associated with retreat. They link this treat with environmental factors (higher temperatures, greater winds that deposit dust on lower portions of glaciers) and with religious factors (divine punishment for human wrong-doing, failure of humans to respect mountain spirits). They describe a variety of economic and extra-economic impacts of this retreat on different spatial, social and temporal scales. Though they face other issues as well (threats of pollution from new mining projects, inadequacy of government services), glacier retreat is their principal concern. Many herders express extreme distress over this unprecedented threat to their livelihoods and communities, though a few propose responses - out-migration, the formation of an association of neighboring communities, development of irrigation works - that could serve as adaptations.

A serologic survey of viral infections in captive ungulates in Turkish zoos.

PubMed

Yeşilbağ, Kadir; Alpay, Gizem; Karakuzulu, Hatice

2011-03-01

Zoos and zoologic gardens make optimal environments for interspecies transmission of viral infections. There are seven zoos and several small zoologic collections in Turkey. This study aimed to determine the current status of viral infections in captive ungulates living in these environments. Blood samples were taken from 163 captive animals from two zoos. There were 39 Cameroon sheep (Ovis ammon f aries), 11 Barbary sheep (Ammotragus lervia), 57 pygmy goats (Capra hircus), 9 Angora goats (Capra hircus), 21 mountain goats (Capra aegagrus-aegagrus), 7 llamas (Lama glama), 8 Persian goitred gazelle (Gazella subgutturosa subgutturosa), 7 Caspian red deer (Cervus elaphus maral), 2 fallow deer (Dama dama), and 2 camels (Camelus dromedarius). Antibodies against bovine viral diarrhea virus (BVDV), bovine herpesvirus-1 (BHV-1), bovine adenoviruses (BAV-1 and -3), parainfluenzavirus 3 (PI-3), and bluetongue viruses (BTV-4 and -9) were investigated using the virus neutralization test, and malignant catarrhal fever (MCF) antibodies were screened by ELISA. All animals were negative for BVDV and BHV-1 antibodies. Seroprevalence of BAV-1, BAV-3, PI-3, BRSV, BT-4, BT-9, and MCF were detected as follows: 46.6%, 60.1%, 0.6%, 7.3%, 1.8%, 1.2%, and 51.6%, respectively. Seroprevalence of BAVs and MCF were more common than all other viruses (P < 0.0001). Ten sheep (37.0%), 48 goats (84.2), and 1 Ilama (14.2%) were the only species positive for MCF antibodies. Prevalence of BRSV and MCF antibodies were found to be significantly higher in goats than in sheep. BTV antibodies were detected both in Cameroon sheep and mountain goats and suggest that zoo animals are at risk for BTV in endemic regions.

Cost-effective control strategies for animal and zoonotic diseases in pastoralist populations.

PubMed

Zinsstag, J; Abakar, M F; Ibrahim, M; Tschopp, R; Crump, L; Bonfoh, B; Schelling, E

2016-11-01

Animal diseases and zoonoses abound among pastoralist livestock, which is composed of cattle, sheep, goats, yak, camels, llamas, reindeer, horses and donkeys. There is endemic and, periodically, epidemic transmission of highly contagious viral and bacterial diseases in Africa, Asia and Latin America. Pastoralist livestock is often multiparasitised with endo- and ectoparasites, as well as being affected by vectorborne viral and protozoal diseases. Pastoral livestock can be a reservoir of such diseases and can also, conversely, be at risk from exposure to wildlife reservoirs. Public and private animal health services currently underperform in almost all pastoral areas due to structural reforms and lack of income, as indicated in assessments of national Veterinary Services by the World Organisation for Animal Health. Control of infectious disease in industrialised countries has been achieved through large-scale public funding of control measures and compensation for culled stock. Such means are not available in pastoralist areas of most low- and middle-income countries (LMICs). While the cost-effectiveness and profitability of the control of animal diseases and zoonoses is less of a consideration for industrialised countries, in the experience of the authors, understanding the economic implications of a control programme is a prerequisite for successful attempts to improve animal health in LMICs. The incremental costs of animal disease control can potentially be shared using crosssector assessments, integrated control, and regional coordination efforts to mitigate transboundary disease risks. In this paper, the authors discuss cost-effective animal disease and zoonoses control in LMICs. It illustrates frameworks and examples of integrated control and cross-sector economics, showing conditions under which these diseases could be controlled in a cost-effective way.

Investigation of a playa lake bed using geophysical electrical methods

NASA Astrophysics Data System (ADS)

Herrmenn, M.; Gurrola, H.; William, R.; Montalvo, R.; Horton, S.; Homberg, J.; Allen, T.; Bribiesca, E.; Lindsey, C.; Anderson, H.; Seshadri, S.; Manns, S.; Hassan, A.; Loren, C.

2005-12-01

The 2005 undergraduate applied geophysical class of Texas Tech University conducted a geophysical survey of a playa lake approximately 10 miles northwest of Lubbock Texas. The playa lake is primarily used as grazing land for two llamas and a hand full of sheep, and has been recently used as a dump for broken down sheds and barrels. Our goal was to model the subsurface of the transition from the playa to plains geology and investigate the possible contamination, of the soil and the data, by the metal dumped at the surface. We conducted our survey with and EM31 and homemade D.C. resistivity and SP equipment that allowed students to grasp the theories more clearly. SP readings were collected using clay pots constructed from terracotta pots and copper tubing purchased at the local hardware store and voltage measurements collected with handle held multi-meters. D.C. resistivity data were collected in a dipole-dipole array using 20 nine volt batteries connected in series with a large enough variable resistor and amp meter to regulate steady current flow. A multi meter was used to collect voltage readings. Wenner array data were collected using a home-made multi-filament cable connected switch box to allow a central user to regulate current and take voltage reading. A map of conductivity produced from a 10 m of EM31 reading show that conductivity anomalies mirror topography. The SP profiles show high values in the playa lake that drop off as we move from the clay rich lake bed to normal grassland. Analysis of both the Dipole-Dipole and Wenner array data support a model with 3 flat layers increasing in resistivity with depth. It appears that these remain flat passing beneath the playa and the playa is eroded into these layers.

Molecular characterization and antibiotic resistance of Enterococcus species from gut microbiota of Chilean Altiplano camelids

PubMed Central

Guerrero-Olmos, Katheryne; Báez, John; Valenzuela, Nicomédes; Gahona, Joselyne; del Campo, Rosa; Silva, Juan

2014-01-01

Background Enterococcus is one of the major human pathogens able to acquire multiple antibiotic-resistant markers as well as virulence factors which also colonize remote ecosystems, including wild animals. In this work, we characterized the Enterococcus population colonizing the gut of Chilean Altiplano camelids without foreign human contact. Material and methods Rectal swabs from 40 llamas and 10 alpacas were seeded in M-Enterococcus agar, and we selected a total of 57 isolates. Species identification was performed by biochemical classical tests, semi-automated WIDER system, mass spectrometry analysis by MALDI-TOF (matrix-assisted laser desorption/ionization with a time-of-flight mass spectrometer), and, finally, nucleotide sequence of internal fragments of the 16S rRNA, rpoB, pheS, and aac(6)-I genes. Genetic diversity was measured by pulsed field gel electrophoresis (PFGE)-SmaI, whereas the antibiotic susceptibility was determined by the WIDER system. Carriage of virulence factors was explored by polymerase chain reaction (PCR). Results Our results demonstrated that the most prevalent specie was Enterococcus hirae (82%), followed by other non–Enterococcus faecalis and non–Enterococcus faecium species. Some discrepancies were detected among the identification methods used, and the most reliable were the rpoB, pheS, and aac(6)-I nucleotide sequencing. Selected isolates exhibited susceptibility to almost all studied antibiotics, and virulence factors were not detected by PCR. Finally, some predominant clones were characterized by PFGE into a diverse genetic background. Conclusion Enterococcus species from the Chilean camelids’ gut microbiota were different from those adapted to humans, and they remained free of antibiotic resistance mechanisms as well as virulence factors. PMID:25405007

Genetic characterization of the oxytocin-neurophysin I gene (OXT) and its regulatory regions analysis in domestic Old and New World camelids

PubMed Central

Ogah, Danlami Moses; Iannaccone, Marco; Erhardt, Georg; Di Stasio, Liliana; Cosenza, Gianfranco

2018-01-01

Oxytocin is a neurohypophysial peptide linked to a wide range of biological functions, including milk ejection, temperament and reproduction. Aims of the present study were a) the characterization of the OXT (Oxytocin-neurophysin I) gene and its regulatory regions in Old and New world camelids; b) the investigation of the genetic diversity and the discovery of markers potentially affecting the gene regulation. On average, the gene extends over 814 bp, ranging between 825 bp in dromedary, 811 bp in Bactrian and 810 bp in llama and alpaca. Such difference in size is due to a duplication event of 21 bp in dromedary. The main regulatory elements, including the composite hormone response elements (CHREs), were identified in the promoter, whereas the presence of mature microRNAs binding sequences in the 3’UTR improves the knowledge on the factors putatively involved in the OXT gene regulation, although their specific biological effect needs to be still elucidated. The sequencing of genomic DNA allowed the identification of 17 intraspecific polymorphisms and 69 nucleotide differences among the four species. One of these (MF464535:g.622C>G) is responsible, in alpaca, for the loss of a consensus sequence for the transcription factor SP1. Furthermore, the same SNP falls within a CpG island and it creates a new methylation site, thus opening future possibilities of investigation to verify the influence of the novel allelic variant in the OXT gene regulation. A PCR-RFLP method was setup for the genotyping and the frequency of the allele C was 0.93 in a population of 71 alpacas. The obtained data clarify the structure of OXT gene in domestic camelids and add knowledge to the genetic variability of a genomic region, which has received little investigation so far. These findings open the opportunity for new investigations, including association studies with productive and reproductive traits. PMID:29608621

Generalists at the interface: Nematode transmission between wild and domestic ungulates.

PubMed

Walker, Josephine G; Morgan, Eric R

2014-12-01

Many parasitic nematode species are generalists capable of infecting multiple host species. The complex life cycle of nematodes, involving partial development outside of the host, facilitates transmission of these parasites between host species even when there is no direct contact between hosts. Infective nematode larvae persist in the environment, and where grazing or water sources are shared ingestion of parasite larvae deposited by different host species is likely. In this paper we examine the extent to which nematode parasite species have been observed in sympatric wild and domestic ungulates. First, using existing host-parasite databases, we describe expected overlap of 412 nematode species between 76 wild and 8 domestic ungulate host species. Our results indicate that host-specific parasites make up less than half of the nematode parasites infecting any particular ungulate host species. For wild host species, between 14% (for common warthog) and 76% (for mouflon) of parasitic nematode species are shared with domestic species. For domestic host species, between 42% (for horse) and 77% (for llamas/alpacas) of parasitic nematode species are shared with wild species. We also present an index of liability to describe the risk of cross-boundary parasites to each host species. We then examine specific examples from the literature in which transmission of nematode parasites between domestic and wild ungulates is described. However, there are many limitations in the existing data due to geographical bias and certain host species being studied more frequently than others. Although we demonstrate that many species of parasitic nematode are found in both wild and domestic hosts, little work has been done to demonstrate whether transmission is occurring between species or whether similar strains circulate separately. Additional research on cross-species transmission, including the use of models and of genetic methods to define strains, will provide evidence to answer this question.

Indigenous Food Systems and Climate Change: Impacts of Climatic Shifts on the Production and Processing of Native and Traditional Crops in the Bolivian Andes.

PubMed

Keleman Saxena, Alder; Cadima Fuentes, Ximena; Gonzales Herbas, Rhimer; Humphries, Debbie L

2016-01-01

Inhabitants of the high-mountain Andes have already begun to experience changes in the timing, severity, and patterning of annual weather cycles. These changes have important implications for agriculture, for human health, and for the conservation of biodiversity in the region. This paper examines the implications of climate-driven changes for native and traditional crops in the municipality of Colomi, Cochabamba, Bolivia. Data were collected between 2012 and 2014 via mixed methods, qualitative fieldwork, including participatory workshops with female farmers and food preparers, semi-structured interviews with local agronomists, and participant observation. Drawing from this data, the paper describes (a) the observed impacts of changing weather patterns on agricultural production in the municipality of Colomi, Bolivia and (b) the role of local environmental resources and conditions, including clean running water, temperature, and humidity, in the household processing techniques used to conserve and sometimes detoxify native crop and animal species, including potato (Solanum sp.), oca (Oxalis tuberosa), tarwi (Lupinus mutabilis), papalisa (Ullucus tuberosus), and charke (llama or sheep jerky). Analysis suggests that the effects of climatic changes on agriculture go beyond reductions in yield, also influencing how farmers make choices about the timing of planting, soil management, and the use and spatial distribution of particular crop varieties. Furthermore, household processing techniques to preserve and detoxify native foods rely on key environmental and climatic resources, which may be vulnerable to climatic shifts. Although these findings are drawn from a single case study, we suggest that Colomi agriculture characterizes larger patterns in what might be termed, "indigenous food systems." Such systems are underrepresented in aggregate models of the impacts of climate change on world agriculture and may be under different, more direct, and more immediate threat from climate change. As such, the health of the food production and processing environments in such systems merits immediate attention in research and practice.

[Ethnographic study of neurological and mental diseases among the Uru-Chipaya peoples of the Andean Altiplano].

PubMed

Carod-Artal, F J; Vázquez-Cabrera, C B

The Uru-Chipaya people are an ethnic group of about 2,500 people, descendants of primitive Andean cultures. Their isolation (they live at an altitude of 4,000 metres in southern Bolivia), their non-written language (Chipaya-Puquina) and their traditional way of life, clothing and customs, which are similar to those used for thousands of years, make this an unusual culture. The aim of our work was to carry out an ethnographic study of the neurological diseases experienced by these people, the way they conceive such disorders and their therapeutic approaches to them. An ethnographic field study was conducted in June 2004. A structured interview was held with a yatiri, or Chipaya healer, to allow classification of the neurological or mental diseases. Epilepsy (tukuri) is interpreted as being a consequence of an evil spirit entering through the nose. Treatment consists in drinking an infusion containing dried powdered butterfly (jesko), birds or curupancho. Achamixi (headache) is common and is treated by drinking the yatiri's fermented urine, herb tea made from the chachacoma plant and by blowing, which is done by the yatiri over the patient's head. Fright, the symptoms of which are similar to those of a post-traumatic stress disorder, is treated by a wilancha, that is, the ritual sacrifice of a llama offered to the Pachamama. Sadness, the cultural equivalent to depression, is treated with infusions made from ayrampo, a plant found in the Andean Altiplano. Psychosis (sumsu), which is treated by means of a wilancha, and mental retardation/static encephalopathy (pustkis), which are considered to be a result of a fright suffered by the mother during pregnancy, also exist. No mention was made of the existence of extrapyramidal or vascular pathologies. The cultural equivalents of certain neurological pathologies (headache, epilepsy, mental retardation, anxiety and depression) are present in this ancestral culture.

Indigenous Food Systems and Climate Change: Impacts of Climatic Shifts on the Production and Processing of Native and Traditional Crops in the Bolivian Andes

PubMed Central

Keleman Saxena, Alder; Cadima Fuentes, Ximena; Gonzales Herbas, Rhimer; Humphries, Debbie L.

2016-01-01

Inhabitants of the high-mountain Andes have already begun to experience changes in the timing, severity, and patterning of annual weather cycles. These changes have important implications for agriculture, for human health, and for the conservation of biodiversity in the region. This paper examines the implications of climate-driven changes for native and traditional crops in the municipality of Colomi, Cochabamba, Bolivia. Data were collected between 2012 and 2014 via mixed methods, qualitative fieldwork, including participatory workshops with female farmers and food preparers, semi-structured interviews with local agronomists, and participant observation. Drawing from this data, the paper describes (a) the observed impacts of changing weather patterns on agricultural production in the municipality of Colomi, Bolivia and (b) the role of local environmental resources and conditions, including clean running water, temperature, and humidity, in the household processing techniques used to conserve and sometimes detoxify native crop and animal species, including potato (Solanum sp.), oca (Oxalis tuberosa), tarwi (Lupinus mutabilis), papalisa (Ullucus tuberosus), and charke (llama or sheep jerky). Analysis suggests that the effects of climatic changes on agriculture go beyond reductions in yield, also influencing how farmers make choices about the timing of planting, soil management, and the use and spatial distribution of particular crop varieties. Furthermore, household processing techniques to preserve and detoxify native foods rely on key environmental and climatic resources, which may be vulnerable to climatic shifts. Although these findings are drawn from a single case study, we suggest that Colomi agriculture characterizes larger patterns in what might be termed, “indigenous food systems.” Such systems are underrepresented in aggregate models of the impacts of climate change on world agriculture and may be under different, more direct, and more immediate threat from climate change. As such, the health of the food production and processing environments in such systems merits immediate attention in research and practice. PMID:26973824

Formatted anti-tumor necrosis factor alpha VHH proteins derived from camelids show superior potency and targeting to inflamed joints in a murine model of collagen-induced arthritis.

PubMed

Coppieters, Ken; Dreier, Torsten; Silence, Karen; de Haard, Hans; Lauwereys, Marc; Casteels, Peter; Beirnaert, Els; Jonckheere, Heidi; Van de Wiele, Christophe; Staelens, Ludovicus; Hostens, Jeroen; Revets, Hilde; Remaut, Erik; Elewaut, Dirk; Rottiers, Pieter

2006-06-01

The advent of tumor necrosis factor (TNF)-blocking drugs has provided rheumatologists with an effective, but highly expensive, treatment for the management of established rheumatoid arthritis (RA). Our aim was to explore preclinically the application of camelid anti-TNF VHH proteins, which are single-domain antigen binding (VHH) proteins homologous to human immunoglobulin V(H) domains, as TNF antagonists in a mouse model of RA. Llamas were immunized with human and mouse TNF, and antagonistic anti-TNF VHH proteins were isolated and cloned for bacterial production. The resulting anti-TNF VHH proteins were recombinantly linked to yield bivalent mouse and human TNF-specific molecules. To increase the serum half-life and targeting properties, an anti-serum albumin anti-TNF VHH domain was incorporated into the bivalent molecules. The TNF-neutralizing potential was analyzed in vitro. Mouse TNF-specific molecules were tested in a therapeutic protocol in murine collagen-induced arthritis (CIA). Disease progression was evaluated by clinical scoring and histologic evaluation. Targeting properties were evaluated by 99mTc labeling and gamma camera imaging. The bivalent molecules were up to 500 times more potent than the monovalent molecules. The antagonistic potency of the anti-human TNF VHH proteins exceeded even that of the anti-TNF antibodies infliximab and adalimumab that are used clinically in RA. Incorporation of binding affinity for albumin into the anti-TNF VHH protein significantly prolonged its serum half-life and promoted its targeting to inflamed joints in the murine CIA model of RA. This might explain the excellent therapeutic efficacy observed in vivo. These data suggest that because of the flexibility of their format, camelid anti-TNF VHH proteins can be converted into potent therapeutic agents that can be produced and purified cost-effectively.

The Rules of Variation Expanded, Implications for the Research on Compatible Genomics.

PubMed

Castro-Chavez, Fernando

2011-05-12

The main focus of this article is to present the practical aspect of the code rules of variation and the search for a second set of genomic rules, including comparison of sequences to understand how to preserve compatible organisms in danger of extinction and how to generate biodiversity. Three new rules of variation are introduced: 1) homologous recombination, 2) a healthy fertile offspring, and 3) comparison of compatible genomes. The novel search in the natural world for fully compatible genomes capable of homologous recombination is explored by using examples of human polymorphisms in the LDLRAP1 gene, and by the production of fertile offspring by crossbreeding. Examples of dogs, llamas and finches will be presented by a rational control of: natural crossbreeding of organisms with compatible genomes (something already happening in nature), the current work focuses on the generation of new varieties after a careful plan. This study is presented within the context of biosemiotics, which studies the processing of information, signaling and signs by living systems. I define a group of organisms having compatible genomes as a single theme: the genomic species or population, able to speak the same molecular language through different accents, with each variety within a theme being a different version of the same book. These studies have a molecular, compatible genetics context. Population and ecosystem biosemiotics will be exemplified by a possible genetic damage capable of causing mutations by breaking the rules of variation through the coordinated patterns of atoms present in the 9/11 World Trade Center contaminated dust (U, Ba, La, Ce, Sr, Rb, K, Mn, Mg, etc.), combination that may be able to overload the molecular quality control mechanisms of the human body. I introduce here the balance of codons in the circular genetic code: 2[1(1)+1(3)+1(4)+4(2)]=2[2(2)+3(4)].

Children Can Learn New Facts Equally Well From Interactive Media Versus Face to Face Instruction

PubMed Central

Kwok, Kristine; Ghrear, Siba; Li, Vivian; Haddock, Taeh; Coleman, Patrick; Birch, Susan A. J.

2016-01-01

Today’s children have more opportunities than ever before to learn from interactive technology, yet experimental research assessing the efficacy of children’s learning from interactive media in comparison to traditional learning approaches is still quite scarce. Moreover, little work has examined the efficacy of using touch-screen devices for research purposes. The current study compared children’s rate of learning factual information about animals during a face-to-face instruction from an adult female researcher versus an analogous instruction from an interactive device. Eighty-six children ages 4 through 8 years (64% male) completed the learning task in either the Face-to-Face condition (n = 43) or the Interactive Media condition (n = 43). In the Learning Phase of the experiment, which was presented as a game, children were taught novel facts about animals without being told that their memory of the facts would be tested. The facts were taught to the children either by an adult female researcher (Face-to-Face condition) or from a pre-recorded female voice represented by a cartoon Llama (Interactive Media condition). In the Testing Phase of the experiment that immediately followed, children’s memory for the taught facts was tested using a 4-option forced-choice paradigm. Children’s rate of learning was significantly above chance in both conditions and a comparison of the rates of learning across the two conditions revealed no significant differences. Learning significantly improved from age 4 to age 8, however, even the preschool-aged children performed significantly above chance, and their performance did not differ between conditions. These results suggest that, interactive media can be equally as effective as one-on-one instruction, at least under certain conditions. Moreover, these results offer support for the validity of using interactive technology to collect data for research purposes. We discuss the implications of these results for children’s learning from interactive media, parental attitudes about interactive technology, and research methods. PMID:27826263

The Rules of Variation Expanded, Implications for the Research on Compatible Genomics

PubMed Central

Castro-Chavez, Fernando

2011-01-01

The main focus of this article is to present the practical aspect of the code rules of variation and the search for a second set of genomic rules, including comparison of sequences to understand how to preserve compatible organisms in danger of extinction and how to generate biodiversity. Three new rules of variation are introduced: 1) homologous recombination, 2) a healthy fertile offspring, and 3) comparison of compatible genomes. The novel search in the natural world for fully compatible genomes capable of homologous recombination is explored by using examples of human polymorphisms in the LDLRAP1 gene, and by the production of fertile offspring by crossbreeding. Examples of dogs, llamas and finches will be presented by a rational control of: natural crossbreeding of organisms with compatible genomes (something already happening in nature), the current work focuses on the generation of new varieties after a careful plan. This study is presented within the context of biosemiotics, which studies the processing of information, signaling and signs by living systems. I define a group of organisms having compatible genomes as a single theme: the genomic species or population, able to speak the same molecular language through different accents, with each variety within a theme being a different version of the same book. These studies have a molecular, compatible genetics context. Population and ecosystem biosemiotics will be exemplified by a possible genetic damage capable of causing mutations by breaking the rules of variation through the coordinated patterns of atoms present in the 9/11 World Trade Center contaminated dust (U, Ba, La, Ce, Sr, Rb, K, Mn, Mg, etc.), combination that may be able to overload the molecular quality control mechanisms of the human body. I introduce here the balance of codons in the circular genetic code: 2[1(1)+1(3)+1(4)+4(2)]=2[2(2)+3(4)]. PMID:21743816

Investigation of the protein osteocalcin of Camelops hesternus: Sequence, structure and phylogenetic implications

NASA Astrophysics Data System (ADS)

Humpula, James F.; Ostrom, Peggy H.; Gandhi, Hasand; Strahler, John R.; Walker, Angela K.; Stafford, Thomas W.; Smith, James J.; Voorhies, Michael R.; George Corner, R.; Andrews, Phillip C.

2007-12-01

Ancient DNA sequences offer an extraordinary opportunity to unravel the evolutionary history of ancient organisms. Protein sequences offer another reservoir of genetic information that has recently become tractable through the application of mass spectrometric techniques. The extent to which ancient protein sequences resolve phylogenetic relationships, however, has not been explored. We determined the osteocalcin amino acid sequence from the bone of an extinct Camelid (21 ka, Camelops hesternus) excavated from Isleta Cave, New Mexico and three bones of extant camelids: bactrian camel ( Camelus bactrianus); dromedary camel ( Camelus dromedarius) and guanaco ( Llama guanacoe) for a diagenetic and phylogenetic assessment. There was no difference in sequence among the four taxa. Structural attributes observed in both modern and ancient osteocalcin include a post-translation modification, Hyp 9, deamidation of Gln 35 and Gln 39, and oxidation of Met 36. Carbamylation of the N-terminus in ancient osteocalcin may result in blockage and explain previous difficulties in sequencing ancient proteins via Edman degradation. A phylogenetic analysis using osteocalcin sequences of 25 vertebrate taxa was conducted to explore osteocalcin protein evolution and the utility of osteocalcin sequences for delineating phylogenetic relationships. The maximum likelihood tree closely reflected generally recognized taxonomic relationships. For example, maximum likelihood analysis recovered rodents, birds and, within hominins, the Homo-Pan-Gorilla trichotomy. Within Artiodactyla, character state analysis showed that a substitution of Pro 4 for His 4 defines the Capra-Ovis clade within Artiodactyla. Homoplasy in our analysis indicated that osteocalcin evolution is not a perfect indicator of species evolution. Limited sequence availability prevented assigning functional significance to sequence changes. Our preliminary analysis of osteocalcin evolution represents an initial step towards a complete character analysis aimed at determining the evolutionary history of this functionally significant protein. We emphasize that ancient protein sequencing and phylogenetic analyses using amino acid sequences must pay close attention to post-translational modifications, amino acid substitutions due to diagenetic alteration and the impacts of isobaric amino acids on mass shifts and sequence alignments.

Mineral, Energy, and Fertilizer Resources of the North Coast of Peru: Perspective from the Santa Rita B Archaeological Site

USGS Publications Warehouse

Brooks, William E.; Kent, Jonathan D.; Willett, Jason C.

2004-01-01

The Santa Rita B archaeological site is in the Chao Valley, approximately 65 km southeast of Trujillo, northern Peru. Location of Santa Rita B at the emergence of several drainages from the Andean cordillera is an important factor in the almost continuous occupation of the site over the past 3,000 years. Mineral resources are abundant throughout the Andes; however, the north coast of Peru was an important center for pre-Columbian mining, metallurgy, and craftsmanship. Success of the Chavin, Moche, Chimu, and other north coast cultures is directly related to the availability and exploitation of mineral and energy resources that include: gold (?silver), as electrum, mainly from placers, and copper from local oxide and carbonate occurrences and from sulfides related to copper porphyry occurrences in the cordillera. An alloy of these three metals is referred to as tumbaga, which is the primary material for Andean metalcraft. Anthracite was used for mirrors by north coast cultures and is available near Rio Chicama, Rio Santa, and east of Santa Rita B. These outcrops are a part of the Alto Chicama, Peru's largest coalfield, which extends from Rio Chicama, in the north, for 200 km southward to Rio Santa. Charcoal from the algorrobo tree and llama dung are considered to be the common pre-Columbian energy sources for cooking and metalwork; however, availability and the higher heat content of anthracite indicate that it was used in metallurgical applications. Bitumen is available from petroleum seeps near Talara, north of the study area, and may have been used as glue or as cement. Hematite, goethite, limonite, and manganese oxides from clay-altered volcanic rock may have provided color and material for ceramics. Guano from the Islas Gua?apes, Chinchas, and Ballestas was used as fertilizer for cotton and other crops.

Generalists at the interface: Nematode transmission between wild and domestic ungulates

PubMed Central

Walker, Josephine G.; Morgan, Eric R.

2014-01-01

Many parasitic nematode species are generalists capable of infecting multiple host species. The complex life cycle of nematodes, involving partial development outside of the host, facilitates transmission of these parasites between host species even when there is no direct contact between hosts. Infective nematode larvae persist in the environment, and where grazing or water sources are shared ingestion of parasite larvae deposited by different host species is likely. In this paper we examine the extent to which nematode parasite species have been observed in sympatric wild and domestic ungulates. First, using existing host–parasite databases, we describe expected overlap of 412 nematode species between 76 wild and 8 domestic ungulate host species. Our results indicate that host-specific parasites make up less than half of the nematode parasites infecting any particular ungulate host species. For wild host species, between 14% (for common warthog) and 76% (for mouflon) of parasitic nematode species are shared with domestic species. For domestic host species, between 42% (for horse) and 77% (for llamas/alpacas) of parasitic nematode species are shared with wild species. We also present an index of liability to describe the risk of cross-boundary parasites to each host species. We then examine specific examples from the literature in which transmission of nematode parasites between domestic and wild ungulates is described. However, there are many limitations in the existing data due to geographical bias and certain host species being studied more frequently than others. Although we demonstrate that many species of parasitic nematode are found in both wild and domestic hosts, little work has been done to demonstrate whether transmission is occurring between species or whether similar strains circulate separately. Additional research on cross-species transmission, including the use of models and of genetic methods to define strains, will provide evidence to answer this question. PMID:25426420

Sobrevivendo a un tsunami: lecciones de Chile, Hawai y Japon

USGS Publications Warehouse

Compilado por Atwater, Brian F.; Cisternas V., Marco; Bourgeois, Joanne; Dudley, Walter C.; Hendley, James W.; Stauffer, Peter H.

1999-01-01

Este folleto contiene historias veridicas que ilustran como sobrevivir, y como no sobrevivir, a un tsunami. Esta publicacion esta dirigida a las personas que viven, trabajan o, simplemente, se divierten a lo largo de las costas que pueden ser afectadas por un tsunami. Tales costas rodean la mayor parte del Oceano Pacifico pero tambien incluyen algunas areas costeras de los Oceanos Atlantico e Indico. Aunque mucha gente llama a los tsunamis 'olas de marea', estos no estan relacionados a las mareas, sino son una serie de olas, o 'tren de olas', generalmente causadas por cambios en el nivel del fondo marino durante los terremotos. Los tsunamis tambien pueden ser generados por la erupcion de volcanes costeros, islas volconicas, deslizamientos submarinos e impactos de grandes meteoritos en el mar. Como sucedio en Sumatra en el 2004, los tsunamis pueden alcanzar alturas de 15 metros, no tan solo en la costa sino tambien kilometros tierra adentro. Los relatos presentados en este folleto fueron seleccionados de entrevistas realizadas a personas que sobrevivieron al tsunami del Oceano Pacifico de 1960. Muchas de estas personas, incluyendo a la enfermera de la foto, se enfrento a las olas generadas a poca distancia, en la costa chilena. En cambio, otros debieron hacer frente al tsunami muchas horas despues, en Hawai y Japon. La mayoria de las entrevistas fueron realizadas a fines de los anos ochenta y en los noventa. Las historias ofrecen una mezcla de lecciones de supervivencia a un tsunami. En algunos casos se presentan las acciones que confiablemente salvaron vidas: poner atencion a los avisos de la naturaleza, abandonar los bienes, dirigirse rapidamente a un sector alto y permanecer alli hasta que el tsunami realmente haya terminado. Otras historias describen como se encontro refugio al subir a construcciones y arboles o flotar sobre desechos, tacticas que tuvieron diferentes resultados y que pueden ser recomendadas solo como actos desesperados de personas atrapadas en sectores bajos.

Nanostructures based on quantum dots for application in promising methods of single- and multiphoton imaging and diagnostics

NASA Astrophysics Data System (ADS)

Nabiev, I. R.

2017-01-01

Molecules recognizing biomarkers of diseases (monoclonal antibodies (monoABs)) are often too large for biomedical applications, and the conditions that are used to bind them with nanolabels lead to disordered orientation of monoABs with respect to the nanoparticle surface. Extremely small nanoprobes, designed via oriented conjugation of quantum dots (QDs) with single-domain antibodies (sdABs) derived from the immunoglobulin of llama and produced in the E. coli culture, have a hydrodynamic diameter less than 12 nm and contain equally oriented sdAB molecules on the surface of each QD. These nanoprobes exhibit excellent specificity and sensitivity in quantitative determination of a small number of cells expressing biomarkers. In addition, the higher diffusion coefficient of sdABs makes it possible to perform immunohistochemical analysis in bulk tissue, inaccessible for conventional monoABs. The necessary conditions for implementing high-quality immunofluorescence diagnostics are a high specificity of labeling and clear differences between the fluorescence of nanoprobes and the autofluorescence of tissues. Multiphoton micros-copy with excitation in the near-IR spectral range, which is remote from the range of tissue autofluorescence excitation, makes it possible to solve this problem and image deep layers in biological tissues. The two-photon absorption cross sections of CdSe/ZnS QDs conjugated with sdABs exceed the corresponding values for organic fluorophores by several orders of magnitude. These nanoprobes provide clear discrimination between the regions of tumor and normal tissues with a ratio of the sdAB fluorescence to the tissue autofluorescence upon two-photon excitation exceeding that in the case of single-photon excitation by a factor of more than 40. The data obtained indicate that the sdAB-QD conjugates used as labels provide the same, or even better, quality as the "gold standard" of immunohistochemical diagnostics. The developed nanoprobes are expected to find wide application in high-efficiency imaging of tumor and multiparameter diagnostics.

Building a world-wide open source community around a software framework: progress, dos, and don'ts

NASA Astrophysics Data System (ADS)

Ibsen, Jorge; Antognini, Jonathan; Avarias, Jorge; Caproni, Alessandro; Fuessling, Matthias; Gimenez, Guillermo; Verma, Khushbu; Mora, Matias; Schwarz, Joseph; Staig, Tomás.

2016-08-01

As we all know too well, building up a collaborative community around a software infrastructure is not easy. Besides recruiting enthusiasts to work as part of it, mostly for free, to succeed you also need to overcome a number of technical, sociological, and, to our surprise, some political hurdles. The ALMA Common Software (ACS) was developed at ESO and partner institutions over the course of more than 10 years. While it was mainly intended for the ALMA Observatory, it was early on thought as a generic distributed control framework. ACS has been periodically released to the public through an LGPL license, which encouraged around a dozen non-ALMA institutions to make use of ACS for both industrial and educational applications. In recent years, the Cherenkov Telescope Array and the LLAMA Observatory have also decided to adopt the framework for their own control systems. The aim of the "ACS Community" is to support independent initiatives in making use of the ACS framework and to further contribute to its development. The Community provides access to a growing network of volunteers eager to develop ACS in areas that are not necessarily in ALMA's interests, and/or were not within the original system scope. Current examples are: support for additional OS platforms, extension of supported hardware interfaces, a public code repository and a build farm. The ACS Community makes use of existing collaborations with Chilean and Brazilian universities, reaching out to promising engineers in the making. At the same time, projects actively using ACS have committed valuable resources to assist the Community's work. Well established training programs like the ACS Workshops are also being continued through the Community's work. This paper aims to give a detailed account of the ongoing (second) journey towards establishing a world-wide open source collaboration around ACS. The ACS Community is growing into a horizontal partnership across a decentralized and diversified group of actors, and we are excited about its technical and human potential.

Modeling Profiles and Signatures of Enrichments

NASA Astrophysics Data System (ADS)

Ali, A.; Qualls, C.; Lucas, S. G.; Lombari, G.; Appenzeller, O.

2014-12-01

Anthropogenic and geochemical enrichment of soils and living matter have been well documented 1, 2, 3.Here we report on geochemical, anthropogenic and biological enrichments with heavy metals in Modern Peru and compared this to Modern and ancient data from New Mexico, USA. We established a signature derived from the quantities of 25 metals in various biological, fossil and soil materials. We also speculate that human adaptation to mercury toxicity may occur in remarkably short time spans during the Holocene. We found mercury concentrations in Modern pigeon feathers and llama wool from free foraging birds and animals in Albuquerque, NM, ranging from 0.006 to 0.019 mg/Kg of tissue. The values for Modern Peru ranged from 22.0 to 556 mg/Kg for the same tissues. We discovered, in 64 million-year-old fossilized plants from New Mexico (Paleocene Nacimiento Formation, San Juan Basin), a mercury concentration of 1.11 mg/Kg of fossil, whereas Modern plant material from the Rio Grande Basin in New Mexico contained no mercury. Profiling of metal content of these samples suggests that mercury is a proxy for anthropogenic rather than geochemical enrichment in the localities we examined. We found no overt signs of mercury toxicity in contemporaneous inhabitants of Huancavelica4, Peru; one of the ten most mercury-polluted places in the world and the mercury concentration in their hair is well below modern admissible levels. However, assessment of their annual scalp hair growth-rate showed marked reduction in growth (~ 5cm/yr) versus ~ 16cm/year for normal scalp hair from other continents4. This is consistent with a toxic effect of heavy metals on human metabolism and especially autonomic nervous system function in Huancavelica, Peru. Contemporaneous anthropogenic activities are known to increase heavy metal content in the biosphere with potentially toxic effects on humans. However, signs of human evolutionary adaptation to such toxins might already be evident in Peru4.

Genetic characterization of the oxytocin-neurophysin I gene (OXT) and its regulatory regions analysis in domestic Old and New World camelids.

PubMed

Pauciullo, Alfredo; Ogah, Danlami Moses; Iannaccone, Marco; Erhardt, Georg; Di Stasio, Liliana; Cosenza, Gianfranco

2018-01-01

Oxytocin is a neurohypophysial peptide linked to a wide range of biological functions, including milk ejection, temperament and reproduction. Aims of the present study were a) the characterization of the OXT (Oxytocin-neurophysin I) gene and its regulatory regions in Old and New world camelids; b) the investigation of the genetic diversity and the discovery of markers potentially affecting the gene regulation. On average, the gene extends over 814 bp, ranging between 825 bp in dromedary, 811 bp in Bactrian and 810 bp in llama and alpaca. Such difference in size is due to a duplication event of 21 bp in dromedary. The main regulatory elements, including the composite hormone response elements (CHREs), were identified in the promoter, whereas the presence of mature microRNAs binding sequences in the 3'UTR improves the knowledge on the factors putatively involved in the OXT gene regulation, although their specific biological effect needs to be still elucidated. The sequencing of genomic DNA allowed the identification of 17 intraspecific polymorphisms and 69 nucleotide differences among the four species. One of these (MF464535:g.622C>G) is responsible, in alpaca, for the loss of a consensus sequence for the transcription factor SP1. Furthermore, the same SNP falls within a CpG island and it creates a new methylation site, thus opening future possibilities of investigation to verify the influence of the novel allelic variant in the OXT gene regulation. A PCR-RFLP method was setup for the genotyping and the frequency of the allele C was 0.93 in a population of 71 alpacas. The obtained data clarify the structure of OXT gene in domestic camelids and add knowledge to the genetic variability of a genomic region, which has received little investigation so far. These findings open the opportunity for new investigations, including association studies with productive and reproductive traits.

Llamas: Large-area microphone arrays and sensing systems

NASA Astrophysics Data System (ADS)

Sanz-Robinson, Josue

Large-area electronics (LAE) provides a platform to build sensing systems, based on distributing large numbers of densely spaced sensors over a physically-expansive space. Due to their flexible, "wallpaper-like" form factor, these systems can be seamlessly deployed in everyday spaces. They go beyond just supplying sensor readings, but rather they aim to transform the wealth of data from these sensors into actionable inferences about our physical environment. This requires vertically integrated systems that span the entirety of the signal processing chain, including transducers and devices, circuits, and signal processing algorithms. To this end we develop hybrid LAE / CMOS systems, which exploit the complementary strengths of LAE, enabling spatially distributed sensors, and CMOS ICs, providing computational capacity for signal processing. To explore the development of hybrid sensing systems, based on vertical integration across the signal processing chain, we focus on two main drivers: (1) thin-film diodes, and (2) microphone arrays for blind source separation: 1) Thin-film diodes are a key building block for many applications, such as RFID tags or power transfer over non-contact inductive links, which require rectifiers for AC-to-DC conversion. We developed hybrid amorphous / nanocrystalline silicon diodes, which are fabricated at low temperatures (<200 °C) to be compatible with processing on plastic, and have high current densities (5 A/cm2 at 1 V) and high frequency operation (cutoff frequency of 110 MHz). 2) We designed a system for separating the voices of multiple simultaneous speakers, which can ultimately be fed to a voice-command recognition engine for controlling electronic systems. On a device level, we developed flexible PVDF microphones, which were used to create a large-area microphone array. On a circuit level we developed localized a-Si TFT amplifiers, and a custom CMOS IC, for system control, sensor readout and digitization. On a signal processing level we developed an algorithm for blind source separation in a real, reverberant room, based on beamforming and binary masking. It requires no knowledge about the location of the speakers or microphones. Instead, it uses cluster analysis techniques to determine the time delays for beamforming; thus, adapting to the unique acoustic environment of the room.

Neutralization Properties of Simian Immunodeficiency Viruses Infecting Chimpanzees and Gorillas

PubMed Central

Barbian, Hannah J.; Decker, Julie M.; Bibollet-Ruche, Frederic; Galimidi, Rachel P.; West, Anthony P.; Learn, Gerald H.; Parrish, Nicholas F.; Iyer, Shilpa S.; Li, Yingying; Pace, Craig S.; Song, Ruijiang; Huang, Yaoxing; Denny, Thomas N.; Mouquet, Hugo; Martin, Loic; Acharya, Priyamvada; Zhang, Baoshan; Kwong, Peter D.; Mascola, John R.; Verrips, C. Theo; Strokappe, Nika M.; Rutten, Lucy; McCoy, Laura E.; Weiss, Robin A.; Brown, Corrine S.; Jackson, Raven; Silvestri, Guido; Connors, Mark; Burton, Dennis R.; Shaw, George M.; Nussenzweig, Michel C.; Bjorkman, Pamela J.; Ho, David D.; Farzan, Michael

2015-01-01

ABSTRACT Broadly cross-reactive neutralizing antibodies (bNabs) represent powerful tools to combat human immunodeficiency virus type 1 (HIV-1) infection. Here, we examined whether HIV-1-specific bNabs are capable of cross-neutralizing distantly related simian immunodeficiency viruses (SIVs) infecting central (Pan troglodytes troglodytes) (SIVcpzPtt) and eastern (Pan troglodytes schweinfurthii) (SIVcpzPts) chimpanzees (n = 11) as well as western gorillas (Gorilla gorilla gorilla) (SIVgor) (n = 1). We found that bNabs directed against the CD4 binding site (n = 10), peptidoglycans at the base of variable loop 3 (V3) (n = 5), and epitopes at the interface of surface (gp120) and membrane-bound (gp41) envelope glycoproteins (n = 5) failed to neutralize SIVcpz and SIVgor strains. In addition, apex V2-directed bNabs (n = 3) as well as llama-derived (heavy chain only) antibodies (n = 6) recognizing both the CD4 binding site and gp41 epitopes were either completely inactive or neutralized only a fraction of SIVcpzPtt strains. In contrast, one antibody targeting the membrane-proximal external region (MPER) of gp41 (10E8), functional CD4 and CCR5 receptor mimetics (eCD4-Ig, eCD4-Igmim2, CD4-218.3-E51, and CD4-218.3-E51-mim2), as well as mono- and bispecific anti-human CD4 (iMab and LM52) and CCR5 (PRO140, PRO140-10E8) receptor antibodies neutralized >90% of SIVcpz and SIVgor strains with low-nanomolar (0.13 to 8.4 nM) potency. Importantly, the latter antibodies blocked virus entry not only in TZM-bl cells but also in Cf2Th cells expressing chimpanzee CD4 and CCR5 and neutralized SIVcpz in chimpanzee CD4+ T cells, with 50% inhibitory concentrations (IC50s) ranging from 3.6 to 40.5 nM. These findings provide new insight into the protective capacity of anti-HIV-1 bNabs and identify candidates for further development to combat SIVcpz infection. PMID:25900654

Vascular provisions for a local utero-ovarian cross-over pathway in new world camelids.

PubMed

Del Campo, M R; Del Campo, C H; Ginther, O J

1996-10-15

The right uterine horn of alpacas causes luteolysis in the right ovary, whereas the left horn causes luteolysis in both ovaries. Female reproductive tracts were studied in 32 adult llamas, 12 adult alpacas, and 21 mid-gestation female fetuses to determine if there is a dichotomy in the vascular anatomy between the 2 sides. Adult tracts were studied by either injection of colored latex into the veins and arteries followed by tissue clearing or by injection of colored fluids during transillumination. Fetal uteri were studied by transillumination. The angioarchitecture of the ovarian vascular pedicle was similar to that reported for ewes. There was no vessel comparable to the middle uterine artery, which is the largest uterine artery in the other farm species. A striking difference from the uterine vascular of other farm species was the presence of a major branch of the right uterine artery that crossed the cranial intercornual area to supply much of the left uterine horn. A corresponding major vein originated from the left horn, crossed the mid-line, and terminated as a branch of the right uterine vein. Thus, the vascular anatomy indicated that much venous blood from the left horn drained to the right side. This was confirmed by injection of colored fluid into a small venous branch at the tip of the left horn. The prominent cross-over vessels were observed in the fetal uteri, and the diameter of the left uterine fetal horn (6.7 +/- 0.6 mm) was greater (P < 0.001) than the diameter of the right horn (5.8 +/- 0.5 mm). The presence of a large cross-over vein traversing from the left horn to the right side is compatible with the hypothesis that the left horn can exert luteolytic control over the corpus luteum in the right ovary through a veno-arterial pathway. The area of veno-arterial transfer of the luteolysin from a vein containing blood from the left horn into an artery supplying the right ovary was not defined in this study. However, the results provide an anatomical basis for functional testing of the cross-over hypothesis and defining the area of venoarterial transfer in camelids.

Pastoralism in Northern Peru during Pre-Hispanic Times: Insights from the Mochica Period (100–800 AD) Based on Stable Isotopic Analysis of Domestic Camelids

PubMed Central

Dufour, Elise; Goepfert, Nicolas; Gutiérrez Léon, Belkys; Chauchat, Claude; Franco Jordán, Régulo; Sánchez, Segundo Vásquez

2014-01-01

Llama (Lama glama) and alpaca (Vicugna pacos) are the only large domesticated animals indigenous to the Americas. Pastoralism occupies a fundamental economic, social and religious role in Andean life. Today, camelid livestock are confined to the ecozone of the puna (above 3,500 masl), while their presence on the Pacific coast during pre-Hispanic times is attested by archaeological skeletal remains. This study aims to document herding practices on the northern Peruvian coast during the Early Intermediate Period (200 BC-600 AD) by gaining insights into diet, location of breeding and mobility of archaeological camelids from the funerary and ritual contexts of two Mochica sites, Uhle Platform in Huacas de Moche and El Brujo. The three first early years and the long-term life histories of the animals were documented by the combined bulk analysis of bone collagen (δ13Ccol and δ15Ncol) and bone structural carbonate (δ13Cbone and δ18Obone) and the serial analysis of structural carbonate of molar tooth enamel (δ13Cenamel and δ18Oenamel). Mochica camelids were bred in the low and/or middle valleys, unlike their modern counterparts, who are restricted to highland puna C3 pastures. Archaeological camelids had diverse and complex life histories, usually with substantial maize foddering. An ontogenetic switch in diet and possible residential mobility during the course of life were identified for some specimens. Although the inference of geographic origin from δ18Obone and δ18Oenamel values was limited because of the lack of understanding of the influence of environmental and biological factors, tooth enamel analysis has great potential for exploring camelid herding practices and Andean pastoralism. Our study suggested that Mochica herders adapted their practices to the difficult lowland environment and that herding practices were varied and not restricted to breeding at higher altitudes. The role of maize in different aspects of the economic life of the Mochicas is also underlined. PMID:24498136

New paleoparasitological investigations from the pre-inca to hispanic contact period in northern Chile.

PubMed

de Souza, Mônica Vieira; da Silva, Lucélia Guedes Ribeiro; Silva-Pinto, Verónica; Mendez-Quiros, Pablo; de Miranda Chaves, Sergio Augusto; Iñiguez, Alena Mayo

2018-02-01

Paleoparasitological studies have demonstrated that changes in environment or culture are reflected in the patterns of parasitic infection diseases in populations worldwide. The advent of agriculture and animal domestication, with its accompanying reduction in human mobility and expanding population involves changes in or emergence of, parasites, the so-called first epidemiological transition. Cultural processes related to territory occupation contribute to both loss and acquisition of parasites. The archaeological site Lluta 57 in the Lluta Valley, Chile, provides a chronology of the transition from the pre-Inca or Late Intermediate Period (LIP), through the Late or Inca Period (LP), to the Hispanic Contact Period (HCP), providing the possibility of evaluating this epidemiological transition. The aim of this study was to conduct a paleoparasitological investigation of to gain insight into the dynamics of parasitism in Lluta people throughout the Inca expansion. Fourteen human coprolites from the three periods were rehydrated, submitted to spontaneous sedimentation, and examined by light microscopy for the presence of intestinal parasite eggs, pollen grains, and micro-remains. Eggs of four parasites: Enterobius vermicularis, Trichostrongylus sp., Trichuris sp., and Eimeria macusaniensis were recovered. Frequency, diversity, and number of parasite eggs per sample increased over the studied time period. Trichostrongylus sp. and E. macusaniensis were recorded in the region for the first time. Enterobius vermicularis eggs, absent in the LIP, were present as a hyper-infection in LP. The presence of E. macusaniensis is likely related to exploitation of llamas, which were used for food and transport and as sacrificial offerings. The paleobotanical analysis revealed ten families of pollen grains, as well as phytoliths and floral remains. In contrast to parasitological results, a diachronic pattern was not detected. Evolution of the settlements, with the advent of larger, more densely populated, villages, could have influenced the emergence and intensification of transmission of parasites in the region. The study showed that the Inca expansion influenced host-parasite-environment relationships in the Lluta Valley. Copyright © 2017 Elsevier B.V. All rights reserved.

Inhibition of HIV Virus by Neutralizing Vhh Attached to Dual Functional Liposomes Encapsulating Dapivirine.

PubMed

Wang, Scarlet Xiaoyan; Michiels, Johan; Ariën, Kevin K; New, Roger; Vanham, Guido; Roitt, Ivan

2016-12-01

Although highly active antiretroviral therapy (HAART) has greatly improved the life expectancy of HIV/AIDS patients, the treatment is not curative. It is a global challenge which fosters an urgent need to develop an effective drug or neutralizing antibody delivery approach for the prevention and treatment of this disease. Due to the low density of envelope spikes with restricted mobility present on the surface of HIV virus, which limit the antibody potency and allow virus mutation and escape from the immune system, it is important for a neutralizing antibody to form bivalent or multivalent bonds with the virus. Liposome constructs could fulfil this need due to the flexible mobility of the membrane with its attached antibodies and the capacity for drug encapsulation. In this study, we evaluated the neutralization activity of a range of liposome formulations in different sizes coated with anti-gp120 llama antibody fragments (Vhhs) conjugated via either non-covalent metal chelation or a covalent linkage. The non-covalent construct demonstrated identical binding affinity to HIV-1 envelope glycoprotein gp120 and neutralizing ability for HIV virus as free Vhh. Although covalently linked Vhh showed significant binding affinity to gp120, it unexpectedly had a lower neutralization potency. This may be due to the comparability in size of the viral and liposome particles restricting the number which can be bound to the liposome surface so involving only a fraction of the antibodies, whereas non-covalently attached antibodies dissociate from the surface after acting with gp120 and free the remainder to bind further viruses. Covalently conjugated Vhh might also trigger the cellular uptake of a liposome-virion complex. To explore the possible ability of the antibody-coated liposomes to have a further function, we encapsulated the hydrophobic antiviral drug dapivirine into both of the non-covalently and covalently conjugated liposome formulations, both of which revealed high efficacy in reducing viral replication in vitro. Thus, dual function liposomes may lead to a novel strategy for the prophylaxis of HIV/AIDS by combining the neutralizing activity of Vhh with antiviral effects of high drug concentrations.

Inhibition of HIV Virus by Neutralizing Vhh Attached to Dual Functional Liposomes Encapsulating Dapivirine

NASA Astrophysics Data System (ADS)

Wang, Scarlet Xiaoyan; Michiels, Johan; Ariën, Kevin K.; New, Roger; Vanham, Guido; Roitt, Ivan

2016-07-01

Although highly active antiretroviral therapy (HAART) has greatly improved the life expectancy of HIV/AIDS patients, the treatment is not curative. It is a global challenge which fosters an urgent need to develop an effective drug or neutralizing antibody delivery approach for the prevention and treatment of this disease. Due to the low density of envelope spikes with restricted mobility present on the surface of HIV virus, which limit the antibody potency and allow virus mutation and escape from the immune system, it is important for a neutralizing antibody to form bivalent or multivalent bonds with the virus. Liposome constructs could fulfil this need due to the flexible mobility of the membrane with its attached antibodies and the capacity for drug encapsulation. In this study, we evaluated the neutralization activity of a range of liposome formulations in different sizes coated with anti-gp120 llama antibody fragments (Vhhs) conjugated via either non-covalent metal chelation or a covalent linkage. The non-covalent construct demonstrated identical binding affinity to HIV-1 envelope glycoprotein gp120 and neutralizing ability for HIV virus as free Vhh. Although covalently linked Vhh showed significant binding affinity to gp120, it unexpectedly had a lower neutralization potency. This may be due to the comparability in size of the viral and liposome particles restricting the number which can be bound to the liposome surface so involving only a fraction of the antibodies, whereas non-covalently attached antibodies dissociate from the surface after acting with gp120 and free the remainder to bind further viruses. Covalently conjugated Vhh might also trigger the cellular uptake of a liposome-virion complex. To explore the possible ability of the antibody-coated liposomes to have a further function, we encapsulated the hydrophobic antiviral drug dapivirine into both of the non-covalently and covalently conjugated liposome formulations, both of which revealed high efficacy in reducing viral replication in vitro. Thus, dual function liposomes may lead to a novel strategy for the prophylaxis of HIV/AIDS by combining the neutralizing activity of Vhh with antiviral effects of high drug concentrations.

Local versus systemic effect of ovulation-inducing factor in the seminal plasma of alpacas

PubMed Central

Ratto, Marcelo H; Huanca, Wilfredo; Singh, Jaswant; Adams, Gregg P

2005-01-01

Background Camelids are induced (reflex) ovulators. We have recently documented the presence of an ovulation-inducing factor (OIF) in the seminal plasma of alpacas and llamas. The objective was to test the hypothesis that OIF exerts its effect via a systemic rather than a local route and that endometrial curettage will enhance the ovulatory response to intrauterine deposition of seminal plasma in alpacas. Methods Female alpacas were assigned randomly to 6 groups (n = 15 to 17 per group) in a 2 × 3 factorial design to test the effect of seminal plasma versus phosphate-buffered saline (PBS) given by intramuscular injection, by intrauterine infusion, or by intrauterine infusion after endometrial curettage. Specifically, alpacas in the respective groups were given 1) 2 ml of alpaca seminal plasma intramuscularly, 2) 2 ml of PBS intramuscularly (negative control group), 3) 2 ml of alpaca seminal plasma by intrauterine infusion, 4) 2 ml of PBS by intrauterine infusion (negative control group), 5) 2 ml of alpaca seminal plasma by intrauterine infusion after endometrial curettage, or 6) 2 ml of PBS by intrauterine infusion after endometrial curettage (negative control group). The alpacas were examined by transrectal ultrasonography to detect ovulation and measure follicular and luteal diameters. Results Intramuscular administration of seminal plasma resulted in a higher ovulation rate than intrauterine administration of seminal plasma (93% versus 41%; P < 0.01), while intrauterine seminal plasma after endometrial curettage was intermediate (67%). None of the saline-treated controls ovulated. The diameter of the CL after treatment-induced ovulation was not affected by the route of administration of seminal plasma. Conclusion We conclude that 1) OIF in seminal plasma effects ovulation via a systemic rather than a local route, 2) disruption of the endometrial mucosa by curettage facilitated the absorption of OIF and increased the ovulatory effect of seminal plasma, and 3) ovulation in alpacas is not associated with a physical stimulation of the genital tract, and 4) the alpaca represents an excellent biological model to evaluate the bioactivity of OIF. PMID:16018817

Molecular epidemiology of feline bordetellosis in two animal shelters in California, USA.

PubMed

Foley, Janet E; Rand, Courtney; Bannasch, Mike J; Norris, Carol R; Milan, Joy

2002-06-25

"Kennel cough" in dogs in animal shelters is readily transmissible, reduces adoption rates, and commonly leads to the euthanasia of affected dogs. In cats, tracheobronchitis, conjunctivitis, and pneumonia have been associated with Bordetella bronchiseptica infection-but most cases of upper-respiratory infection (URI) probably are caused by herpesvirus and calicivirus, and many B. bronchiseptica culture-positive cats are clinically normal. Our prospective observational study was undertaken to document the contribution of B. bronchiseptica to disease in cats and dogs from two animal shelters undergoing outbreaks of canine kennel cough, to evaluate whether cross-species transmission might have occurred, and to determine if the presence of infected cats represented a risk to dogs. Clinically defined cases of kennel cough in dogs and URI in cats were investigated in two shelters by calculating clinical-disease incidence, alveolar-lavage cytological examination, bacterial and viral cultures, antibiotic-susceptibility testing, and molecular fingerprinting by pulsed-field gel electrophoresis. In a 40-cat and 40-dog "no-kill" shelter, the prevalences of culture positivity were 47% for B. bronchiseptica and 36% for calicivirus at the same time as two resident dogs demonstrated clinical cough. When no dogs had kennel cough 3 months later, 10% of cats were B. bronchiseptica-culture-positive and 63% calicivirus positive. In a large traditional shelter, the incidence of kennel cough in dogs increased over 12 weeks to a maximum of 19 cases/week/120 dogs, during which time the culture prevalence was 23% for B. bronchiseptica in dogs and 47% in cats. Three to 6 months before the kennel-cough epidemic, no dogs or cats were B. bronchiseptica positive. Very little genetic variability was detected in isolates from these shelters; all isolates except one corresponded to a single strain type which was identical to the pattern in a vaccine used in these shelters. Isolates from other cats, a horse, a llama, and a sea otter were genetically distinct from the shelter isolates. There was widespread resistance to cephalosporins and ampicillin, but low or no resistance to amoxicillin/clavulanate, trimethoprim-sulfamethoxazole, tetracycline, and enrofloxacin. Greater percent resistance was observed in the traditional shelter than in the no-kill shelter and feline isolates were more likely to be resistant than canine isolates.

Revision of the Middle American clade of the ant genus Stenamma Westwood (Hymenoptera, Formicidae, Myrmicinae)

PubMed Central

Branstetter, Michael G.

2013-01-01

Abstract Stenamma is a cryptic “leaf-litter” ant genus that occurs in mesic forest habitats throughout the Holarctic region, Central America, and part of northwestern South America (Colombia and Ecuador). The genus was thought to be restricted primarily to the temperate zone, but recent collecting efforts have uncovered a large radiation of Neotropical forms, which rival the Holarctic species in terms of morphological and behavioral diversity. By inferring a broad-scale molecular phylogeny of Stenamma, Branstetter (2012) showed that all Neotropical species belong to a diverse Middle American clade (MAC), and that this clade is sister to an almost completely geographically separated Holarctic clade (HOC). Here, the Middle American clade of Stenamma is revised to recognize 40 species, of which 33 are described as new. Included in the revision are a key to species based on the worker caste, and for each species where possible, descriptions and images of workers and queens, images of males, information on geographic distribution, descriptions of intraspecific variation, and notes on natural history. Several species groups are defined, but the majority of species remain unassigned due to a lack of diagnostic morphological character states for most molecular clades. The following species are redescribed: Stenamma alas Longino, Stenamma diversum Mann, Stenamma expolitum Smith, Stenamma felixi Mann, Stenamma huachucanum Smith, Stenamma manni Wheeler, and Stenamma schmidti Menozzi. The following are described as new: Stenamma andersoni sp. n., Stenamma atribellum sp. n., Stenamma brujita sp. n., Stenamma callipygium sp. n., Stenamma catracho sp. n., Stenamma connectum sp. n., Stenamma crypticum sp. n., Stenamma cusuco sp. n., Stenamma excisum sp. n., Stenamma expolitico sp. n., Stenamma hojarasca sp. n., Stenamma ignotum sp. n., Stenamma lagunum sp. n., Stenamma llama sp. n., Stenamma leptospinum sp. n., Stenamma lobinodus sp. n., Stenamma longinoi sp. n., Stenamma maximon sp. n., Stenamma megamanni sp. n., Stenamma monstrosum sp. n., Stenamma muralla sp. n., Stenamma nanozoi sp. n., Stenamma nonotch sp. n., Stenamma ochrocnemis sp. n., Stenamma pelophilum sp. n., Stenamma picopicucha sp. n., Stenamma saenzae sp. n., Stenamma sandinista sp. n., Stenamma stictosomum sp. n., Stenamma tiburon sp. n., Stenamma tico sp. n., Stenamma vexator sp. n., and Stenamma zelum sp. n. Although many of the newly defined species consist of challenging species complexes, this study establishes a robust baseline that will guide future work on the systematics of MAC Stenamma. The total global diversity of Stenamma now includes 84 extant species. PMID:23794874

A Novel ¹¹¹In-Labeled Anti-Prostate-Specific Membrane Antigen Nanobody for Targeted SPECT/CT Imaging of Prostate Cancer.

PubMed

Chatalic, Kristell L S; Veldhoven-Zweistra, Joke; Bolkestein, Michiel; Hoeben, Sander; Koning, Gerben A; Boerman, Otto C; de Jong, Marion; van Weerden, Wytske M

2015-07-01

Prostate-specific membrane antigen (PSMA) is overexpressed in prostate cancer (PCa) and a promising target for molecular imaging and therapy. Nanobodies (single-domain antibodies, VHH) are the smallest antibody-based fragments possessing ideal molecular imaging properties, such as high target specificity and rapid background clearance. We developed a novel anti-PSMA Nanobody (JVZ-007) for targeted imaging and therapy of PCa. Here, we report on the application of the (111)In-radiolabeled Nanobody for SPECT/CT imaging of PCa. A Nanobody library was generated by immunization of a llama with 4 human PCa cell lines. Anti-PSMA Nanobodies were captured by biopanning on PSMA-overexpressing cells. JVZ-007 was selected for evaluation as an imaging probe. JVZ-007 was initially produced with a c-myc-hexahistidine (his) tag allowing purification and detection. The c-myc-his tag was subsequently replaced by a single cysteine at the C terminus, allowing site-specific conjugation of chelates for radiolabeling. JVZ-007-c-myc-his was conjugated to 2-(4-isothiocyanatobenzyl)-diethylenetriaminepentaacetic acid (p-SCN-DTPA) via the lysines, whereas JVZ-007-cys was conjugated to maleimide-DTPA via the C-terminal cysteine. PSMA targeting was analyzed in vitro by cell-binding experiments using flow cytometry, autoradiography, and internalization assays with various PCa cell lines and patient-derived xenografts (PDXs). The targeting properties of radiolabeled Nanobodies were evaluated in vivo in biodistribution and SPECT/CT imaging experiments, using nude mice bearing PSMA-positive PC-310 and PSMA-negative PC-3 tumors. JVZ-007 was successfully conjugated to DTPA for radiolabeling with (111)In at room temperature. (111)In-JVZ007-c-myc-his and (111)In-JVZ007-cys internalized in LNCaP cells and bound to PSMA-expressing PDXs and, importantly, not to PSMA-negative PDXs and human kidneys. Good tumor targeting and fast blood clearance were observed for (111)In-JVZ-007-c-myc-his and (111)In-JVZ-007-cys. Renal uptake of (111)In-JVZ-007-c-myc-his was initially high but was efficiently reduced by coinjection of gelofusine and lysine. The replacement of the c-myc-his tag by the cysteine contributed to a further reduction of renal uptake without loss of targeting. PC-310 tumors were clearly visualized by SPECT/CT with both tracers, with low renal uptake (<4 percentage injected dose per gram) for (111)In-JVZ-007-cys already at 3 h after injection. We developed an (111)In-radiolabeled anti-PSMA Nanobody, showing good tumor targeting, low uptake in nontarget tissues, and low renal retention, allowing excellent SPECT/CT imaging of PCa within a few hours after injection. © 2015 by the Society of Nuclear Medicine and Molecular Imaging, Inc.

The rocks and fossils of my school… as educational and cientific divulgation resources

NASA Astrophysics Data System (ADS)

Canas, Ana; Lopes, Francisco; Guerreiro, Catarina; Reis, Raquel; Carneiro, Amanda

2017-04-01

In a city, on walls, pavements, street furniture and monuments, it is normal to find rocks with fossils and curious features, full of interesting stories. This geological and paleontological heritage, usually ignored by the majority of citizens, has an enormous potential for use in science; education; conservation; communication; tourism and leisure. The fact that the walls and floors in the Silves Secondary School are covered with different types of rocks with peculiar characteristics and full of an enormous fossiliferous diversity, gave us the opportunity to explore the potential of this geodiversity in a school context. The main objectives defined in this project were: to promote the geodiversity present in the school building as an educational (sciences' teaching) and communication (scientific literacy and divulgation) resource; to diversify the educational materials and resources available in the school; to involve students in a concrete project, with a practical and multidisciplinary approach; to stimulate students' interest for science and knowledge; to promote the education of more observing, conscientious and respectful citizens of the natural and cultural heritage. In the execution of the project, we highlight the following activities: theoretical and practical work sessions (bibliographical research, field work, analysis and discussion of results, evaluation, ...); inventory of rocks and fossils ("cartography" of the school building, written and photographic record, identification of rocks and fossils, ...); selection of places of interest; production and compilation of contents - texts, photographs, diagrams and illustrations (students of both science and technology, and arts courses); production of a digital guide with sections for students of different levels of education, teachers and other interested parties (available on the school website); production of identifying and informative contents for boards in the places of interest; edition and graphic design of the digital guide and informative boards (students of the graphic design course); practical classes, guided tours and other promotion and divulgation initiatives. In the end, with this project, students, teachers and the rest of the educational community, will regard rocks and fossils differently. In the next school year (2017/2018), we intend to carry the work forward, extending it to other schools in Silves county. Some of the papers that inspired our project: -Cachada, M., Santos, A., Alfaro, E., Silva, C. S. (2012). Experiencias de aprovechamiento educativo y turístico de recursos geológicos en las ciudades de Huelva, Sevilla y Córdoba. Comunicaciones XVII Simposio sobre Enseñanza de la Geología. Universidad de Huelva, 64-70 -Fuertes, I., Calzada, E., Llamas, T., Tejerina, A., Crespo, M., Pereiras, L., Crespo, T., Domínguez, L., Cabezas, L. (2016). Lugares de interés geoeducativo en el medio urbano. Potencialidad de las ciudades para la enseñanza de Geología. Enseñanza de las Ciencias de la Tierra, Vol. 24 (2), 195-201 -Rodrigues, L., Agostinho, M., Manteigas, R. (2014). Geologia e Paleontologia Urbanas - potencialidades e aplicações em três cidades do Algarve. Comunicações Geológicas, 101, Especial III: 1359-1363 -Silva, C. M., Cachão, M. (1998). "Paleontologia Urbana": percursos citadinos de interpretação e educação (paleo)ambiental. Actas V Congresso Nacional de Geologia, Comunicações Instituto Geológico e Mineiro, Lisboa, 84 (2), H33-H37

Close to the Sky

NASA Astrophysics Data System (ADS)

2007-11-01

Today, a new ALMA outreach and educational book was publicly presented to city officials of San Pedro de Atacama in Chile, as part of the celebrations of the anniversary of the Andean village. ESO PR Photo 50a/07 ESO PR Photo 50a/07 A Useful Tool for Schools Entitled "Close to the sky: Biological heritage in the ALMA area", and edited in English and Spanish by ESO in Chile, the book collects unique on-site observations of the flora and fauna of the ALMA region performed by experts commissioned to investigate it and to provide key initiatives to protect it. "I thank the ALMA project for providing us a book that will surely be a good support for the education of children and youngsters of San Pedro de Atacama. Thanks to this publication, we expect our rich flora and fauna to be better known. I invite teachers and students to take advantage of this educational resource, which will be available in our schools", commented Ms. Sandra Berna, the Mayor of San Pedro de Atacama, who was given the book by representatives of the ALMA global collaboration project. Copies of the book 'Close to the sky' will be donated to all schools in the area, as a contribution to the education of students and young people in northern Chile. "From the very beginning of the project, ALMA construction has had a firm commitment to environment and local culture, protecting unique flora and fauna species and preserving old estancias belonging to the Likan Antai culture," said Jacques Lassalle, who represented ALMA at the hand-over. "Animals like the llama, the fox or the condor do not only live in the region where ALMA is now being built, but they are also key elements of the ancient Andean constellations. In this sense they are part of the same sky that will be explored by ALMA in the near future." ESO PR Photo 50c/07 ESO PR Photo 50c/07 Presentation of the ALMA book The ALMA Project is a giant, international observatory currently under construction on the high-altitude Chajnantor site in Chile. ALMA will be composed initially of 66 high-precision telescopes, operating at wavelengths of 0.3 to 9.6 mm. The ALMA antennas will be electronically combined and will provide astronomical observations which are equivalent to those from a single large telescope of tremendous size and resolution. Chajnantor was selected as the ideal spot for ALMA, following several years of atmospheric and meteorology studies. The high elevation, stable atmosphere, and low humidity make it one of the best locations in the world for radio astronomy. To protect the outstanding conditions of Chajnantor, the Government of Chile declared a major portion of the area a scientific reserve. The publication is available in PDF format. It is the second book on ALMA for the general public, following the previous launch of "Footprints in the Desert", also available on the Internet in PDF format in Spanish. ALMA is a partnership between Europe, East Asia and North America in cooperation with the Republic of Chile. It is funded in Europe by ESO, in East Asia by the National Institutes of Natural Sciences of Japan in cooperation with the Academia Sinica in Taiwan and in North America by the U.S. National Science Foundation in cooperation with the National Research Council of Canada. ALMA construction and operations are led on behalf of Europe by ESO, on behalf of East Asia by the National Astronomical Observatory of Japan and on behalf of North America by the National Radio Astronomy Observatory, which is managed by Associated Universities, Inc.

Foot & Mouth Disease & Ulcerative/Vesicular Rule-outs: Challenges Encountered in Recent Outbreaks

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hullinger, P

2008-01-28

Foot and mouth disease (FMD) is a highly infectious and contagious viral disease affecting bovidae (cattle, zebus, domestic buffaloes, yaks), sheep, goats, swine, all wild ruminants and suidae. Camelidae (camels, dromedaries, llamas, vicunas) have low susceptibility. Foot and mouth disease is caused by a RNS virus of the family Picornaviridae, genus Aphthovirus. There are seven immunologically distinct serotypes: A, O, C, SAT1, SAT2, SAT3, Asia 1. Foot and mouth disease causes significant economic loss both to countries who manage it as an endemic disease (with or without vaccination), as well as those FMD free countries which may become infected. Themore » mortality rate is low in adult animals, but often higher in young due to myocarditis. Foot and mouth disease is endemic in parts of Asia, Africa, the Middle East and South America (sporadic outbreaks in free areas). The Office of International Epizootics (OIE), also referred to the World Organization for Animal Health maintains an official list of free countries and zones.1 The OIE Terrestrial Code (Chapter 2.2.10) provides detailed information on the categories of freedom that can be allocated to a country as well as guidelines for the surveillance for foot and mouth disease (Appendix 3.8.7). In short, countries may be completely free of FMD, free with vaccination or infected with foot and mouth disease virus (FMDV). Source of FMDV include incubating and clinically affected animals with virus present in breath, saliva, faeces, urine, milk and semen. In experimental settings virus has been detected in milk several days before the onset of clinical signs2. Additional sources of virus are meat and by-products in which pH has remained above 6.0 as well as persistently infected carrier animals. Carrier animals may include cattle and water buffalo; convalescent animals and exposed vaccinates (virus persists in the oropharynx for up to 30 months in cattle or longer in buffalo, 9 months in sheep). Pigs do not become carriers. It has been shown that the African Cape buffalo are the major maintenance host of SAT serotypes. FMDV transmission can occur by either direct or indirect contact. Indirect transmission can occur via contaminated animate vectors (humans, etc.), inanimate vectors (vehicles, implements) or airborne transmission. Indirect disease transmission via animate or inanimate vectors can play a major role in disease transmission. Good biosecurity can significantly reduce this type of transmission. Airborne transmission is often debated and is known to be serotype and species specific as well as require specific environmental conditions to occur. Airborne transmission is favored in temperate zones and has been postulated to occur over distances of up to 60 km overland and 300 km by sea. Foot and mouth disease virus is an unenveloped virus which is preserved by refrigeration and freezing and progressively inactivated by temperatures above 50 C. FMDV is highly sensitive to pH change and is inactivated by pH < 6.0 or > 9.0. There are many disinfectants which are effective against FMDV including sodium hydroxide (2%), sodium carbonate (4%), and citric acid (0.2%). FMDV is resistant to iodophores, quaternary ammonium compounds, hypochlorite and phenol, especially in the presence of organic matter. The virus can survive in lymph nodes and bone marrow at neutral pH, but is destroyed in muscle when is pH < 6.0 i.e. after rigor mortis. FMDV can persist in contaminated feed/commodities and the environment for over to 1 month, depending on the temperature and pH conditions. The incubation period for FMD is 2-14 days. Animals transition through latent (infected but not infectious), subclinically infected (infectious but lacking clinical signs) clinically infected and recovered disease states. In cattle clinical signs include pyrexia, reluctance to eat, bruxism, drooling, lameness, treading or stamping of the feet and decreased milk production. Most clinical signs are related to the development and subsequent rupturing of vesicles at the coronary band and in the oral cavity. Vesicles and ulcerations can also occur on the mammary gland. Recovery in adult animals usually occurs in 8-15 days. Clinical signs for most serotypes are less dramatic in sheep and goats. Swine can develop very severe coronary band lesions and high mortality in piglets has been observed. One of the challenges of diagnosing FMD is that it may be clinically similar to several other vesicular or ulcerative diseases. FMD is clinically indistinguishable from Vesicular stomatitis, Swine vesicular disease and Vesicular exanthema of swine. It may also resemble Bovine viral diarrhea, Mucosal disease, Infectious bovine rhinotracheitis, Bluetongue, Bovine papular stomatitis, Bovine mammillitis and Rinderpest.« less

Faconnage de la surface de nanoparticules pour la delivrance de genes

NASA Astrophysics Data System (ADS)

Fortier, Charles

Gene vectorization has become an essential tool in the field of biotechnology. It is also found at the heart of research and development of gene therapies, a specific goal of which is to deliver genetic material in the nuclei of target cells. As the DNA to be delivered is fragile, most vectorization techniques include the use of nanoobjects that are able to encapsulate and protect this delicate cargo. These nanoobjects also facilitate gene transport to the nuclei of target cells whereby its expression can occur. These nanovectors are made from varied materials that include polymers. Particularly, polyplexes form a class of nanovectors that derive from the complexation between DNA and a positively charged polymer. The works presented here are focused on the role of the interface between a model polyplex and the surrounding cellular and physiological environments during the process of gene delivery or transfection. More specifically, in the framework of oncology, systemic delivery has been investigated to enable polyplex accumulation in tumor tissue as well as in ensuing metastases. The stakes are twofold here: it is relevant to decorate polyplexes both with ligands that enable the specific targeting of cancer cells in order to deliver genes at the right place and with a complete surface coating enabling them to circulate more freely in the bloodstream while limiting their elimination by the mononuclear phagocyte system. In a first study, we set up a system to tether ligands at the surface of polyplexes. We based our works on the E/Kcoil tandem: two peptides that enable a stable, oriented and specific tethering between two entities by auto assembly in solution. On the one side, we chose a ligand: a chimeric protein corresponding to the Ecoil peptide fused with the epidermal growth factor (Ecoil-EGF); and on the other side, we engineered a Kcoil peptide-decorated polyplex. In that endeavor, a cationic polymer - branched polyethylenimine (bPEI) - was functionalized with a moiety reactive to the Kcoil peptide. Polyplexes prepared with this activated bPEI could then be decorated at their surface by the covalent grafting of Kcoil. This system enabled us to fine-tune the ligand surface density in order to study its influence on polyplex uptake by cancer-derived EGF receptor (EGFR)-over-expressing A431 cells. Thus, we evidenced a switch effect between the EGFR-specific polyplex uptake rate by A431 cells and the amount of ligand on polyplexes. Then, we carried out a second study that was focused on the electrostatic coating of polyplexes with carboxymethylated dextran (CMD). The objective was to draw relations between the coating structure - defined by the charge density and the molecular weight of the CMD used - and its function relative to the biological environment. We prepared and characterized a chemical library of 26 closely related CMDs by the controlled carboxymethylation of 4 dextrans differing by their molecular weight. A simple and robust electrostatic coating procedure was set up in order to carry out a systematic study of the CMD library. Each coated polyplex formulation was tested on decoupled relevant constituents of the physiological milieu: small ions, DNases, plasma proteins, red blood cells, and target cells. We observed monotonic trends between both parameters and DNA encapsulation. To our great surprise, those observations did not correlate with DNA protection against DNase challenging, the latter correlating with target cells reporter gene expression. Polyplex stability was evaluated at physiological salt concentration; the results of which indicated that both studied parameters had a jointed influence in preventing polyplex aggregation. At the surface of polyplexes, this influence translated in terms of steric and lateral stabilization rather than in terms of surface charge (zeta potential). Conversely, the latter turned out to be a strong predictor of the ability of coated polyplexes to prevent both opsonization and erythrocytes aggregation. Thus, we exposed the complementarity of several relevant interfacial characteristics for the use of polyplexes in vivo, whereby broadening our understanding of the tailoring of the surface of polyplexes with a hydrophilic polyanion. Next, we envisioned the design of a transfection agent comprising of the E/Kcoil system together with a cationic peptide (RRRRRHHHHHC or R5H5) directly grafted on a dextran backbone. This approach was developed in order to take advantage of the growing collection of Ecoil-tagged polypeptides that were being developed and characterized in our group: especially one derived from the vascular endothelial growth factor (Ecoil-VEGF), and the other from the endosomolytic peptide GALA (Ecoil-GALA). In that endeavor, two dextran-peptides conjugates were prepared and characterized: Dextran-R5H5 and Dextran-R5H5,-Kcoil. Their abilities to encapsulate DNA and to form nanoparticles were confirmed. Subsequently, in vitro transfection assays demonstrated no significant gain from controls: Dextran-R5H5 could transfect cells with no greater efficacy than that of linear PEI. We then equipped our polyplexes with Ecoil-GALA in order to facilitate endosomal escape; however we could not evidence any additional gain regarding that latter point or with respect to overall reporter gene expression. In another side study, we evaluated the utility of polyanions such as CMD in transient gene expression (TGE) for the production of r-proteins in bioreactors. We had previously observed that one of the beneficial effects of polyplex coating with CMD was related to the complexation of excess polycation, which had been linked to the cytotoxicity observed during polyplex-mediated transfection. In parallel, we confirmed that the direct use of coated polyplexes did reduce toxicity during transfection, albeit with decreased r-protein yield. We thus conjectured that the addition of polyanion after transfection could attenuate excess polycation-associated toxicity while maintaining high TGE yields, in order to increase the overall r-protein yield. A library of 8 varied polyanions was used in order to test this hypothesis. Although the delay before addition was explored along with the amount of polyanion, no reliable procedure could be brought out. The last study that we carried out dealt with DNA immunization of animals for the generation of antibodies. The hydrodynamic injection of naked plasmid DNA is currently being used in that endeavor for small mammals such as mice. This procedure is effective yet not efficient as it proves lethal for one in four animals in our facility. The direct use of PEI/DNA polyplexes is also limited for it proves even riskier for the animal. These two techniques feature a systemic injection that renders them hardly scalable to larger animals such as llamas, a particularly relevant species in the field of antibodies generation. We first took on setting up a lyophilisation procedure for our most promising coated polyplex formulation for further use in mice. Successful lyophilizing enabled us to prepare in advance polyplexes to be rehydrated in a highly concentrated yet isotonic fashion that was better adapted for systemic injection. We selected appropriate formulations based on the knowledge gained from the analysis of our CMD library. A first assay was carried out with polyplexes coated with CMD70-60, the most stable formulation that was found to comply with every criterion for systemic injection. Corresponding results indicated that no lethal side effect arose, however the resulting gene expression in mice was undetectable. We then carried out a second round of injections on the same mice cohort, with two less stable formulations: a CMD 40-30 coating that still complied with every physiological criterion was chosen for systemic injection; and a CMD40-03 coating that featured better in vitro transfection performances but did not otherwise comply with every physiological criterion was injected subcutaneously. The subcutaneous injection proved lethal to every animal, which suggests that our previous in vitro systematic analysis bears predictive power over in vivo performances.